U.S. patent application number 12/288775 was filed with the patent office on 2009-05-28 for novel genes encoding proteins having prognostic, diagnostic preventive, therapeutic, and other uses.
This patent application is currently assigned to Millennium Pharmaceuticals, Inc.. Invention is credited to Thomas M. Barnes, Christopher C. Fraser, Andrew D.J. Goodearl, Douglas A. Holtzman, Mehran Khodadoust, Susan J. Kirst, Sean A. McCarthy, Paul S. Myers, John D. Sharp, Nicholas Wrighton.
Application Number | 20090136959 12/288775 |
Document ID | / |
Family ID | 23301576 |
Filed Date | 2009-05-28 |
United States Patent
Application |
20090136959 |
Kind Code |
A1 |
McCarthy; Sean A. ; et
al. |
May 28, 2009 |
Novel genes encoding proteins having prognostic, diagnostic
preventive, therapeutic, and other uses
Abstract
The invention provides isolated nucleic acids encoding a variety
of proteins having diagnostic, preventive, therapeutic, and other
uses. These nucleic and proteins are useful for diagnosis,
prevention, and therapy of a number of human and other animal
disorders. The invention also provides antisense nucleic acid
molecules, expression vectors containing the nucleic acid molecules
of the invention, host cells into which the expression vectors have
been introduced, and non-human transgenic animals in which a
nucleic acid molecule of the invention has been introduced or
disrupted. The invention still further provides isolated
polypeptides, fusion polypeptides, antigenic peptides and
antibodies. Diagnostic, screening, and therapeutic methods using
compositions of the invention are also provided. The nucleic acids
and polypeptides of the present invention are useful as modulating
agents in regulating a variety of cellular processes.
Inventors: |
McCarthy; Sean A.; (San
Diego, CA) ; Fraser; Christopher C.; (Lexington,
MA) ; Sharp; John D.; (Arlington, MA) ;
Barnes; Thomas M.; (Brookline, MA) ; Kirst; Susan
J.; (Brookline, MA) ; Myers; Paul S.; (Jamaica
Plain, MA) ; Wrighton; Nicholas; (Winchester, MA)
; Goodearl; Andrew D.J.; (Natick, MA) ; Holtzman;
Douglas A.; (Jamaica Plain, MA) ; Khodadoust;
Mehran; (Brookline, MA) |
Correspondence
Address: |
MILLENNIUM PHARMACEUTICALS, INC.
40 Landsdowne Street
CAMBRIDGE
MA
02139
US
|
Assignee: |
Millennium Pharmaceuticals,
Inc.
|
Family ID: |
23301576 |
Appl. No.: |
12/288775 |
Filed: |
October 23, 2008 |
Related U.S. Patent Documents
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Application
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Filing Date |
Patent Number |
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11491318 |
Jul 21, 2006 |
7459530 |
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12288775 |
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09766511 |
Jan 19, 2001 |
7160694 |
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11491318 |
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09578063 |
May 24, 2000 |
6764677 |
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09766511 |
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09333159 |
Jun 14, 1999 |
7033780 |
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09578063 |
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09596194 |
Jun 16, 2000 |
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09766511 |
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09342364 |
Jun 29, 1999 |
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09596194 |
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09608452 |
Jun 30, 2000 |
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09766511 |
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09393996 |
Sep 10, 1999 |
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09608452 |
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09345680 |
Jun 30, 1999 |
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09766511 |
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Current U.S.
Class: |
435/6.11 ;
435/320.1; 435/325; 435/375; 435/402; 435/69.1; 435/7.2; 436/86;
530/350; 530/387.9; 536/23.1 |
Current CPC
Class: |
C07K 14/47 20130101 |
Class at
Publication: |
435/6 ; 536/23.1;
435/320.1; 435/325; 530/350; 530/387.9; 435/69.1; 435/7.2; 435/375;
435/402; 436/86 |
International
Class: |
C12Q 1/68 20060101
C12Q001/68; C12N 15/11 20060101 C12N015/11; C12N 15/00 20060101
C12N015/00; C12N 5/06 20060101 C12N005/06; C07K 14/00 20060101
C07K014/00; G01N 33/00 20060101 G01N033/00; C07K 1/00 20060101
C07K001/00; C07K 16/18 20060101 C07K016/18; C12P 21/04 20060101
C12P021/04; G01N 33/53 20060101 G01N033/53 |
Claims
1. An isolated nucleic acid molecule selected from the group
consisting of: a) a nucleic acid molecule having a nucleotide
sequence which is at least 90% identical to the nucleotide sequence
of any of SEQ ID NOs: 1, 2, 11, 12, 21, 22, 31, 32, 41, 42, 51, 52,
61, 62, 71, 72, 81, 82, and the nucleotide sequence of any of the
clones deposited as ATCC.RTM. Accession numbers 207185, 207221,
PTA-147, PTA-425, and PTA-424, or a complement thereof; b) a
nucleic acid molecule comprising at least 15 nucleotide residues
and having a nucleotide sequence identical to at least 15
consecutive nucleotide residues of any of SEQ ID NOs: 1, 2, 11, 12,
21, 22, 31, 32, 41, 42, 51, 52, 61, 62, 71, 72, 81, 82, and the
nucleotide sequence of any of the clones deposited as ATCC.RTM.
Accession numbers 207185, 207221, PTA-147, PTA-425, and PTA-424, or
a complement thereof; c) a nucleic acid molecule which encodes a
polypeptide comprising the amino acid sequence of any of SEQ ID
NOs: 3-8, 13-18, 23-28, 33-38, 43, 53-55, 63-65, 73, and 83-85, and
the amino acid sequence encoded by the nucleotide sequence of any
of the clones deposited as ATCC.RTM. Accession numbers 207185,
207221, PTA-147, PTA-425, and PTA-424; d) a nucleic acid molecule
which encodes a fragment of a polypeptide comprising the amino acid
sequence of any of SEQ ID NOs: 3-8, 13-18, 23-28, 33-38, 43, 53-55,
63-65, 73, and 83-85 and the amino acid sequence encoded by the
nucleotide sequence of any of the clones deposited as ATCC.RTM.
Accession numbers 207185, 207221, PTA-147, PTA-425, and PTA-424,
wherein the fragment comprises at least 10 consecutive amino acid
residues of any of SEQ ID NOs: 3-8, 13-18, 23-28, 33-38, 43, 53-55,
63-65, 73, and 83-85 and the amino acid sequence encoded by the
nucleotide sequence of any of the clones deposited as ATCC.RTM.
Accession numbers 207185, 207221, PTA-147, PTA-425, and PTA-424; e)
a nucleic acid molecule which encodes a fragment of a polypeptide
comprising the amino acid sequence of any of SEQ ID NOs: 3-8,
13-18, 23-28, 33-38, 43, 53-55, 63-65, 73, and 83-85 and the amino
acid sequence encoded by the nucleotide sequence of any of the
clones deposited as ATCC.RTM. Accession numbers 207185, 207221,
PTA-147, PTA-425, and PTA-424, wherein the fragment comprises
consecutive amino acid residues corresponding to at least half of
the full length of any of SEQ ID NOs: 3, 13, 23, 33, 43, 53, 63,
73, and 83 and the amino acid sequence encoded by the nucleotide
sequence of any of the clones deposited as ATCC.RTM. Accession
numbers 207185, 207221, PTA-147, PTA-425, and PTA-424; and f) a
nucleic acid molecule which encodes a naturally occurring allelic
variant of a polypeptide comprising the amino acid sequence of any
of SEQ ID NOs: 3-8, 13-18, 23-28, 33-38, 43, 53-55, 63-65, 73, and
83-85, wherein the nucleic acid molecule hybridizes with a nucleic
acid molecule consisting of the nucleotide sequence of any of SEQ
ID NOs: 1, 2, 11, 12, 21, 22, 31, 32, 41, 42, 51, 52, 61, 62, 71,
72, 81, 82, and the nucleotide sequence of any of the clones
deposited as ATCC.RTM. Accession numbers 207185, 207221, PTA-147,
PTA-425, and PTA-424, or a complement thereof under stringent
conditions.
2. The isolated nucleic acid molecule of claim 1, which is selected
from the group consisting of: a) a nucleic acid having the
nucleotide sequence of any of SEQ ID NOs: 1, 2, 11, 12, 21, 22, 31,
32, 41, 42, 51, 52, 61, 62, 71, 72, 81, 82, and the nucleotide
sequence of any of the clones deposited as ATCC.RTM. Accession
numbers 207185, 207221, PTA-147, PTA-425, and PTA-424, or a
complement thereof; and b) a nucleic acid molecule which encodes a
polypeptide having the amino acid sequence of any of SEQ ID NOs:
3-8, 13-18, 23-28, 33-38, 43, 53-55, 63-65, 73, and 83-85 and the
amino acid sequence encoded by the nucleotide sequence of any of
the clones deposited as ATCC.RTM. Accession numbers 207185, 207221,
PTA-147, PTA-425, and PTA-424, or a complement thereof.
3. The nucleic acid molecule of claim 1, further comprising vector
nucleic acid sequences.
4. The nucleic acid molecule of claim 1 further comprising nucleic
acid sequences encoding a heterologous polypeptide.
5. A host cell which contains the nucleic acid molecule of claim
1.
6. The host cell of claim 5 which is a mammalian host cell.
7. A non-human mammalian host cell containing the nucleic acid
molecule of claim 1.
8. An isolated polypeptide selected from the group consisting of:
a) a fragment of a polypeptide comprising the amino acid sequence
of any of SEQ ID NOs: 3-8, 13-18, 23-28, 33-38, 43, 53-55, 63-65,
73, and 83-85 and the amino acid sequence encoded by the nucleotide
sequence of any of the clones deposited as ATCC.RTM. Accession
numbers 207185, 207221, PTA-147, PTA-425, and PTA-424; b) a
naturally occurring allelic variant of a polypeptide comprising the
amino acid sequence of any of SEQ ID NOs: 3-8, 13-18, 23-28, 33-38,
43, 53-55, 63-65, 73, and 83-85, wherein the polypeptide is encoded
by a nucleic acid molecule which hybridizes with a nucleic acid
molecule consisting of the nucleotide sequence of any of SEQ ID
NOs: 1, 2, 11, 12, 21, 22, 31, 32, 41, 42, 51, 52, 61, 62, 71, 72,
81, 82, and the nucleotide sequence of any of the clones deposited
as ATCC.RTM. Accession numbers 207185, 207221, PTA-147, PTA-425,
and PTA-424, or a complement thereof under stringent conditions;
and c) a polypeptide which is encoded by a nucleic acid molecule
comprising a nucleotide sequence which is at least 90% identical to
a nucleic acid consisting of the nucleotide sequence of any of SEQ
ID NOs: 1, 2, 11, 12, 21, 22, 31, 32, 41, 42, 51, 52, 61, 62, 71,
72, 81, 82, and the nucleotide sequence of any of the clones
deposited as ATCC.RTM. Accession numbers 207185, 207221, PTA-147,
PTA-425, and PTA-424, or a complement thereof.
9. The isolated polypeptide of claim 8 having the amino acid
sequence of any of SEQ ID NOs: 3-8, 13-18, 23-28, 33-38, 43, 53-55,
63-65, 73, and 83-85 and the amino acid sequence encoded by the
nucleotide sequence of any of the clones deposited as ATCC.RTM.
Accession numbers 207185, 207221, PTA-147, PTA-425, and
PTA-424.
10. The polypeptide of claim 8, wherein the amino acid sequence of
the polypeptide further comprises heterologous amino acid
residues.
11. An antibody which selectively binds with the polypeptide of
claim 8.
12. A method for producing a polypeptide selected from the group
consisting of: a) a polypeptide comprising the amino acid sequence
of any of SEQ ID NOs: 3-8, 13-18, 23-28, 33-38, 43, 53-55, 63-65,
73, and 83-85 and the amino acid sequence encoded by the nucleotide
sequence of any of the clones deposited as ATCC.RTM. Accession
numbers 207185, 207221, PTA-147, PTA-425, and PTA-424; b) a
polypeptide comprising a fragment of the amino acid sequence of any
of SEQ ID NOs: 3-8, 13-18, 23-28, 33-38, 43, 53-55, 63-65, 73, and
83-85 and the amino acid sequence encoded by the nucleotide
sequence of any of the clones deposited as ATCC.RTM. Accession
numbers 207185, 207221, PTA-147, PTA-425, and PTA-424, wherein the
fragment comprises at least 10 contiguous amino acids of any of SEQ
ID NOs: 3-8, 13-18, 23-28, 33-38, 43, 53-55, 63-65, 73, and 83-85
and the amino acid sequence encoded by the nucleotide sequence of
any of the clones deposited as ATCC.RTM. Accession numbers 207185,
207221, PTA-147, PTA-425, and PTA-424; and c) a naturally occurring
allelic variant of a polypeptide comprising the amino acid sequence
of any of SEQ ID NOs: 3-8, 13-18, 23-28, 33-38, 43, 53-55, 63-65,
73, and 83-85, wherein the polypeptide is encoded by a nucleic acid
molecule which hybridizes with a nucleic acid molecule consisting
of the nucleotide sequence of any of SEQ ID NOs: 1, 2, 11, 12, 21,
22, 31, 32, 41, 42, 51, 52, 61, 62, 71, 72, 81, 82, and the
nucleotide sequence of any of the clones deposited as ATCC.RTM.
Accession numbers 207185, 207221, PTA-147, PTA-425, and PTA-424, or
a complement thereof under stringent conditions; the method
comprising culturing the host cell of claim 5 under conditions in
which the nucleic acid molecule is expressed.
13. A method for detecting the presence of a polypeptide of claim 8
in a sample, comprising: a) contacting the sample with a compound
which selectively binds with a polypeptide of claim 8; and b)
determining whether the compound binds with the polypeptide in the
sample.
14. The method of claim 13, wherein the compound which binds with
the polypeptide is an antibody.
15. A method for detecting the presence of a nucleic acid molecule
of claim 1 in a sample, comprising the steps of: a) contacting the
sample with a nucleic acid probe or primer which selectively
hybridizes with the nucleic acid molecule; and b) determining
whether the nucleic acid probe or primer binds with a nucleic acid
molecule in the sample.
16. The method of claim 15, wherein the sample comprises mRNA
molecules and is contacted with a nucleic acid probe.
17. A method for identifying a compound which binds with a
polypeptide of claim 8 comprising the steps of: a) contacting a
polypeptide, or a cell expressing a polypeptide of claim 8 with a
test compound; and b) determining whether the polypeptide binds
with the test compound.
18. The method of claim 17, wherein the binding of the test
compound to the polypeptide is detected by a method selected from
the group consisting of: a) detection of binding by direct
detecting of test compound/polypeptide binding; b) detection of
binding using a competition binding assay; c) detection of binding
using an assay for an activity characteristic of the
polypeptide.
19. A method for modulating the activity of a polypeptide of claim
8 comprising contacting a polypeptide or a cell expressing a
polypeptide of claim 8 with a compound which binds with the
polypeptide in a sufficient concentration to modulate the activity
of the polypeptide.
20. A method for identifying a compound which modulates the
activity of a polypeptide of claim 8, comprising: a) contacting a
polypeptide of claim 8 with a test compound; and b) determining the
effect of the test compound on the activity of the polypeptide to
thereby identify a compound which modulates the activity of the
polypeptide.
Description
CROSS-REFERENCE TO RELATED APPLICATIONS
[0001] This application is a divisional application of U.S. patent
application Ser. No. 11/491,318, filed Jul. 21, 2006, pending,
which is a divisional application of U.S. patent application Ser.
No. 09/766,511, filed Jan. 19, 2001, now U.S. Pat. No.
7,160,694.
[0002] U.S. patent application Ser. No. 09/766,511 is a
continuation-in-part of U.S. patent application Ser. No.
09/578,063, filed on May 24, 2000, now U.S. Pat. No. 6,764,677,
which is a continuation-in-part of U.S. patent application Ser. No.
09/333,159, filed on Jun. 14, 1999, now U.S. Pat. No.
7,033,780.
[0003] U.S. patent application Ser. No. 09/766,511 is also a
continuation-in-part of U.S. patent application Ser. No.
09/596,194, filed on Jun. 16, 2000, now abandoned, which is a
continuation-in-part of U.S. patent application Ser. No.
09/342,364, filed on Jun. 29, 1999, now abandoned.
[0004] U.S. patent application Ser. No. 09/766,511 is also a
continuation-in-part of U.S. patent application Ser. No.
09/608,452, filed on Jun. 30, 2000, now abandoned, which is a
continuation-in-part of U.S. patent application Ser. No.
09/393,996, filed on Sep. 10, 1999, now abandoned.
[0005] U.S. patent application Ser. No. 09/766,511 is also a
continuation-in-part of U.S. patent application Ser. No.
09/345,680, filed on Jun. 30, 1999, now abandoned.
[0006] The contents of each of the applications cross-referenced in
this section are incorporated into this disclosure by
reference.
STATEMENT REGARDING FEDERAL RESEARCH SUPPORT
[0007] Not Applicable
REFERENCE TO MICROFICHE APPENDIX
[0008] Not Applicable
BACKGROUND OF THE INVENTION
[0009] The molecular bases underlying many human and animal
physiological states (e.g., diseased and homeostatic states of
various tissues) remain unknown. Nonetheless, it is well understood
that these states result from interactions among the proteins and
nucleic acids present in the cells of the relevant tissues. In the
past, the complexity of biological systems overwhelmed the ability
of practitioners to understand the molecular interactions giving
rise to normal and abnormal physiological states. More recently,
though, the techniques of molecular biology, transgenic and null
mutant animal production, computational biology, and
pharmacogenomics have enabled practitioners to discern the role and
importance of individual genes and proteins in particular
physiological states.
[0010] Knowledge of the sequences and other properties of genes
(particularly including the portions of genes encoding proteins)
and the proteins encoded thereby enables the practitioner to design
and screen agents which will affect, prospectively or
retrospectively, the physiological state of an animal tissue in a
favorable way. Such knowledge also enables the practitioner, by
detecting the levels of gene expression and protein production, to
diagnose the current physiological state of a tissue or animal and
to predict such physiological states in the future. This knowledge
furthermore enables the practitioner to identify and design
molecules which bind with the polynucleotides and proteins, in
vitro, in vivo, or both.
[0011] Many secreted proteins, for example, cytokines and cytokine
receptors, play a vital role in the regulation of cell growth, cell
differentiation, and a variety of specific cellular responses. A
number of medically useful proteins, including erythropoietin,
granulocyte-macrophage colony stimulating factor, human growth
hormone, and various interleukins, are secreted proteins. Thus, an
important goal in the design and development of new therapies is
the identification and characterization of secreted and
transmembrane proteins and the genes which encode them.
[0012] Many secreted proteins are receptors which bind a ligand and
transduce an intracellular signal, leading to a variety of cellular
responses. The identification and characterization of such a
receptor enables one to identify both the ligands which bind to the
receptor and the intracellular molecules and signal transduction
pathways associated with the receptor, permitting one to identify
or design modulators of receptor activity, e.g., receptor agonists
or antagonists and modulators of signal transduction.
SUMMARY OF THE INVENTION
[0013] The present invention is based, at least in part, on the
discovery of human cDNA molecules which encode proteins which are
herein designated TANGO 273, TANGO 325, TANGO 364, TANGO 405, and
M019 (M019 is synonymous with TANGO 533). These proteins, fragments
thereof, derivatives thereof, and variants thereof are collectively
referred to herein as the polypeptides of the invention or the
proteins of the invention. Nucleic acid molecules encoding
polypeptides of the invention are collectively referred to as
nucleic acids of the invention.
[0014] The nucleic acids and polypeptides of the present invention
are useful as modulating agents for regulating a variety of
cellular processes. Accordingly, in one aspect, the present
invention provides isolated nucleic acid molecules encoding a
polypeptide of the invention or a biologically active portion
thereof. The present invention also provides nucleic acid molecules
which are suitable as primers or hybridization probes for the
detection of nucleic acids encoding a polypeptide of the
invention.
[0015] The invention also includes fragments of any of the nucleic
acids described herein wherein the fragment retains a biological or
structural function by which the full-length nucleic acid is
characterized (e.g., an activity, an encoded protein, or a binding
capacity). The invention furthermore includes fragments of any of
the nucleic acids described herein wherein the fragment has a
nucleotide sequence sufficiently (e.g., 50%, 60%, 70%, 80%, 85%,
90%, 95%, 98%, or 99% or greater) identical to the nucleotide
sequence of the corresponding full-length nucleic acid that it
retains a biological or structural function by which the
full-length nucleic acid is characterized (e.g., an activity, an
encoded protein, or a binding capacity).
[0016] The invention also includes fragments of any of the
polypeptides described herein wherein the fragment retains a
biological or structural function by which the full-length
polypeptide is characterized (e.g., an activity or a binding
capacity). The invention furthermore includes fragments of any of
the polypeptides described herein wherein the fragment has an amino
acid sequence sufficiently (e.g., 50%, 60%, 70%, 80%, 85%, 90%,
95%, 98%, or 99% or greater) identical to the amino acid sequence
of the corresponding full-length polypeptide that it retains a
biological or structural function by which the full-length
polypeptide is characterized (e.g., an activity or a binding
capacity).
[0017] The invention also features nucleic acid molecules which are
at least 40% (or 50%, 60%, 70%, 80%, 90%, 95%, or 98%) identical to
the nucleotide sequence of any of SEQ ID NOs: 1, 2, 11, 12, 21, 22,
31, 32, 41, 42, 51, 52, 61, 62, 71, 72, 81, and 82, the human TANGO
273 nucleotide sequence of the cDNA insert of a clone deposited on
Apr. 2, 1999 with the American Type Culture Collection.RTM.
(ATCC.RTM.) as accession no. 207185, the murine TANGO 273
nucleotide sequence of the cDNA insert of a clone deposited on Apr.
2, 1999 with ATCC.RTM. as accession no. 207221, the human TANGO 325
nucleotide sequence of the cDNA insert of a clone deposited on May
28, 1999 with ATCC.RTM. as accession no. PTA-147, the human TANGO
364 nucleotide sequence of the cDNA insert of a clone deposited on
Jul. 23, 1999 with ATCC.RTM. as accession no. PTA-425, the human
TANGO 405 nucleotide sequence of the cDNA insert of a clone
deposited on Jul. 23, 1999 with ATCC.RTM. as accession no. PTA-424,
or a complement thereof. These deposited nucleotide sequences are
hereafter individually and collectively referred to as "the
nucleotide sequence of any of the clones deposited as ATCC.RTM.
Accession numbers 207185, 207221, PTA-147, PTA-425, and
PTA-424."
[0018] The invention features nucleic acid molecules which include
a fragment of at least 15 (25, 40, 60, 80, 100, 150, 200, 250, 300,
350, 400, 450, 500, 600, 700, 800, 900, 1000, 1200, 1400, 1600,
1800, 2000, 2200, 2400, 2600, 2800, 3000, or 3500 or more)
consecutive nucleotide residues of any of SEQ ID NOs: 1, 2, 11, 12,
21, 22, 31, 32, 41, 42, 51, 52, 61, 62, 71, 72, 81, 82, and the
nucleotide sequence of any of the clones deposited as ATCC.RTM.
Accession numbers 207185, 207221, PTA-147, PTA-425, and PTA-424, or
a complement thereof.
[0019] The invention also features nucleic acid molecules which
include a nucleotide sequence encoding a protein having an amino
acid sequence that is at least 50% (or 60%, 70%, 80%, 90%, 95%, or
98%) identical to the amino acid sequence of any of SEQ ID NOs:
3-8, 13-18, 23-28, 33-38, 43, 53-55, 63-65, 73, and 83-85, or the
amino acid sequence encoded by the nucleotide sequence of any of
the clones deposited as ATCC.RTM. Accession numbers 207185, 207221,
PTA-147, PTA-425, and PTA-424, or a complement thereof.
[0020] In certain embodiments, the nucleic acid molecules have the
nucleotide sequence of any of SEQ ID NOs: 1, 2, 11, 12, 21, 22, 31,
32, 41, 42, 51, 52, 61, 62, 71, 72, 81, 82, and the nucleotide
sequence of any of the clones deposited as ATCC.RTM. Accession
numbers 207185, 207221, PTA-147, PTA-425, and PTA-424.
[0021] Also within the invention are nucleic acid molecules which
encode a fragment of a polypeptide having the amino acid sequence
of any of SEQ ID NOs: 3-8, 13-18, 23-28, 33-38, 43, 53-55, 63-65,
73, and 83-85, the fragment including at least 10 (12, 15, 20, 25,
30, 40, 50, 75, 100, 125, 150, 200, 250, 300, 400, 500, 750, 1000
or more) consecutive amino acid residues of any of SEQ ID NOs: 3-8,
13-18, 23-28, 33-38, 43, 53-55, 63-65, 73, and 83-85.
[0022] The invention includes nucleic acid molecules which encode a
naturally occurring allelic variant of a polypeptide comprising the
amino acid sequence of any of SEQ ID NOs: 3-8, 13-18, 23-28, 33-38,
43, 53-55, 63-65, 73, and 83-85, wherein the nucleic acid molecule
hybridizes under stringent conditions to a nucleic acid molecule
having a nucleic acid sequence of any of SEQ ID NOs: 1, 2, 11, 12,
21, 22, 31, 32, 41, 42, 51, 52, 61, 62, 71, 72, 81, 82, and the
nucleotide sequence of any of the clones deposited as ATCC.RTM.
Accession numbers 207185, 207221, PTA-147, PTA-425, and PTA-424, or
a complement thereof.
[0023] Also within the invention are isolated polypeptides or
proteins having an amino acid sequence that is at least about 50%,
preferably 60%, 75%, 90%, 95%, or 98% identical to the amino acid
sequence of any of SEQ ID NOs: 3-8, 13-18, 23-28, 33-38, 43, 53-55,
63-65, 73, and 83-85.
[0024] Also within the invention are isolated polypeptides or
proteins which are encoded by a nucleic acid molecule having a
nucleotide sequence that is at least about 40%, preferably 50%,
60%, 75%, 85%, or 95% identical to the nucleic acid sequence
encoding any of SEQ ID NOs: 3-8, 13-18, 23-28, 33-38, 43, 53-55,
63-65, 73, and 83-85, and isolated polypeptides or proteins which
are encoded by a nucleic acid molecule consisting of the nucleotide
sequence which hybridizes under stringent hybridization conditions
to a nucleic acid molecule having the nucleotide sequence of any of
SEQ ID NOs: 1, 2, 11, 12, 21, 22, 31, 32, 41, 42, 51, 52, 61, 62,
71, 72, 81, 82, and the nucleotide sequence of any of the clones
deposited as ATCC.RTM. Accession numbers 207185, 207221, PTA-147,
PTA-425, and PTA-424.
[0025] Also within the invention are polypeptides which are
naturally occurring allelic variants of a polypeptide that includes
the amino acid sequence of any of SEQ ID NOs: 3-8, 13-18, 23-28,
33-38, 43, 53-55, 63-65, 73, and 83-85, wherein the polypeptide is
encoded by a nucleic acid molecule which hybridizes under stringent
conditions to a nucleic acid molecule having the nucleotide
sequence of any of SEQ ID NOs: 1, 2, 11, 12, 21, 22, 31, 32, 41,
42, 51, 52, 61, 62, 71, 72, 81, 82, and the nucleotide sequence of
any of the clones deposited as ATCC.RTM. Accession numbers 207185,
207221, PTA-147, PTA-425, and PTA-424, or a complement thereof.
[0026] The invention also features nucleic acid molecules that
hybridize under stringent conditions to a nucleic acid molecule
having the nucleotide sequence of any of SEQ ID NOs: 1, 2, 11, 12,
21, 22, 31, 32, 41, 42, 51, 52, 61, 62, 71, 72, 81, 82, and the
nucleotide sequence of any of the clones deposited as ATCC.RTM.
Accession numbers 207185, 207221, PTA-147, PTA-425, and PTA-424, or
a complement thereof. In other embodiments, the nucleic acid
molecules are at least 15 (25, 40, 60, 80, 100, 150, 200, 250, 300,
350, 400, 450, 500, 600, 700, 800, 900, 1000, 1200, 1400, 1600,
1800, 2000, 2200, 2400, 2600, 2800, 3000, or 3500 or more)
nucleotides in length and hybridize under stringent conditions to a
nucleic acid molecule having the nucleotide sequence of any of SEQ
ID NOs: 1, 2, 11, 12, 21, 22, 31, 32, 41, 42, 51, 52, 61, 62, 71,
72, 81, 82, and the nucleotide sequence of any of the clones
deposited as ATCC.RTM. Accession numbers 207185, 207221, PTA-147,
PTA-425, and PTA-424, or a complement thereof. In some embodiments,
the isolated nucleic acid molecules encode a cytoplasmic,
transmembrane, extracellular, or other domain of a polypeptide of
the invention. In other embodiments, the invention provides an
isolated nucleic acid molecule which is antisense to the coding
strand of a nucleic acid of the invention.
[0027] Another aspect of the invention provides vectors, e.g.,
recombinant expression vectors, comprising a nucleic acid molecule
of the invention. In another embodiment, the invention provides
isolated host cells, e.g., mammalian or non-mammalian cells,
containing such a vector or a nucleic acid of the invention. The
invention also provides methods for producing a polypeptide of the
invention by culturing, in a suitable medium, a host cell of the
invention containing a recombinant expression vector encoding a
polypeptide of the invention such that the polypeptide of the
invention is produced.
[0028] Another aspect of this invention features isolated or
recombinant proteins and polypeptides of the invention. Preferred
proteins and polypeptides possess at least one biological activity
possessed by the corresponding naturally-occurring human
polypeptide. An activity, a biological activity, and a functional
activity of a polypeptide of the invention refers to an activity
exerted by a protein or polypeptide of the invention on a
responsive cell as determined in vivo, or in vitro, according to
standard techniques. Such activities can be a direct activity, such
as an association with or an enzymatic activity exerted on a second
protein or an indirect activity, such as a cellular processes
mediated by interaction of the protein with a second protein.
[0029] TANGO 273 protein mediates one or more physiological
responses of cells to bacterial infection, e.g., by mediating one
or more of detection of bacteria in a tissue in which it is
expressed, movement of cells with relation to sites of bacterial
infection, production of biological molecules which inhibit
bacterial infection, and production of biological molecules which
alleviate cellular or other physiological damage wrought by
bacterial infection. TANGO 273, a transmembrane protein, is also
involved in transmembrane signal transduction, and therefore
mediates transmission of signals between the extracellular and
intracellular environments of cells. TANGO 273 mediates regulation
of cell growth and proliferation, endocytosis, activation of
respiratory burst, and other physiological processes triggered by
transmission of a signal via a protein with which TANGO 273
interacts. The compositions and methods of the invention can
therefore be used to prevent, diagnose, and treat disorders
involving one or more physiological activities mediated by TANGO
273 protein.
[0030] As an additional example, TANGO 325 polypeptides, nucleic
acids, and modulators thereof modulate growth, proliferation,
survival, differentiation, and activity of human tissues such as
vascular endothelium, including aortic endothelium, other heart
tissues, placenta, liver, kidney, and pancreas tissues. Thus, TANGO
325 polypeptides, nucleic acids, and modulators thereof can
therefore be used to prevent, diagnose, and treat disorders
involving one or more physiological activities mediated by TANGO
325 protein in tissues in which it is expressed. Such activities
include, for example, modulation of cardiac contractility and
vasomotor tone, modulation of leukocyte extravasation, sensing
physiological signals by the endocrine system, modulating growth,
development, maintenance, and regeneration of neurons, and the
like.
[0031] TANGO 364, compounds which modulate its activity,
expression, or both, and compounds (e.g., antibodies) which bind
with TANGO 364 (collectively "TANGO 364-related molecules") exhibit
the ability to affect one or more of growth, proliferation,
survival, differentiation, activity, morphology, and
movement/migration of, for example, human fetal and adult skin
cells and tissue. Furthermore, TANGO 364 is involved in modulating
cell-to-cell adhesion, tissue and extracellular matrix invasivity
of cells, infectivity of cells by pathogens (e.g., bacteria and
viruses), endocrine signaling processes, tissue developmental and
organizational processes, and the like. Thus, TANGO 364-related
molecules can be used to prognosticate, prevent, diagnose, or treat
one or more disorders associated with these physiological
processes.
[0032] TANGO 405, compounds which modulate its activity,
expression, or both, and compounds (e.g., antibodies) which bind
with TANGO 405 (collectively "TANGO 405-related molecules")
modulate one or more of growth, proliferation, survival,
differentiation, activity, morphology, and movement/migration of
human lymphocytes and bone marrow cells and tissues. As described
herein, TANGO 405 is involved in activation of leukocytes,
including modulating one or more of growth, proliferation,
survival, differentiation, activity, morphology,
movement/migration, and other cellular processes by which
leukocytes are characterized. TANGO 405 is involved in disorders
associated with aberrant activation of leukocytes, including both
auto-immune disorders and disorders related to inappropriate
activity or activation of leukocytes and disorders related to
uncontrolled proliferation of leukocytes.
[0033] M019 protein, compounds which modulate its activity,
expression, or both, and compounds (e.g., antibodies) which bind
with M019 (collectively "M019-related molecules") exhibit the
ability to affect growth, proliferation, survival, differentiation,
and activity of adipose tissue cells.
[0034] In one embodiment, a polypeptide of the invention has an
amino acid sequence sufficiently identical to a polypeptide of the
invention or to an identified domain thereof. As used herein, the
term "sufficiently identical" refers to a first amino acid or
nucleotide sequence which contains a sufficient or minimum number
of identical or equivalent (e.g., with a similar side chain) amino
acid residues or nucleotides to a second amino acid or nucleotide
sequence such that the first and second amino acid or nucleotide
sequences have a common domain and/or common functional activity.
For example, amino acid or nucleotide sequences which contain a
common domain having about 65% identity, preferably 75% identity,
more preferably 85%, 95%, or 98% identity are defined herein as
sufficiently identical.
[0035] In one embodiment, the isolated polypeptide of the invention
lacks both a transmembrane and a cytoplasmic domain. In another
embodiment, the polypeptide lacks both a transmembrane domain and a
cytoplasmic domain and is soluble under physiological
conditions.
[0036] The polypeptides of the present invention, or biologically
active portions thereof, can be operably linked with a heterologous
amino acid sequence to form fusion proteins. The invention further
features antibody substances that specifically bind a polypeptide
of the invention, such as monoclonal or polyclonal antibodies,
antibody fragments, and single-chain antibodies. In addition, the
polypeptides of the invention or biologically active portions
thereof can be incorporated into pharmaceutical compositions, which
optionally include pharmaceutically acceptable carriers. These
antibody substances can be made, for example, by providing the
polypeptide of the invention to an immunocompetent vertebrate and
thereafter harvesting blood or serum from the vertebrate.
[0037] In another aspect, the present invention provides methods
for detecting the presence of the activity or expression of a
polypeptide of the invention in a biological sample by contacting
the biological sample with an agent capable of detecting an
indicator of activity such that the presence of activity is
detected in the biological sample.
[0038] In another aspect, the invention provides methods for
modulating activity of a polypeptide of the invention comprising
contacting a cell with an agent that modulates (inhibits or
enhances) the activity or expression of a polypeptide of the
invention such that activity or expression in the cell is
modulated. In one embodiment, the agent is an antibody that
specifically binds with a polypeptide of the invention.
[0039] In another embodiment, the agent modulates expression of a
polypeptide of the invention by modulating transcription, splicing,
or translation of an mRNA encoding a polypeptide of the invention.
In yet another embodiment, the agent is a nucleic acid molecule
having a nucleotide sequence that is antisense with respect to the
coding strand of an mRNA encoding a polypeptide of the
invention.
[0040] The present invention also provides methods of treating a
subject having a disorder characterized by aberrant activity of a
polypeptide of the invention or aberrant expression of a nucleic
acid of the invention by administering an agent which is a
modulator of the activity of a polypeptide of the invention or a
modulator of the expression of a nucleic acid of the invention to
the subject. In one embodiment, the modulator is a protein of the
invention. In another embodiment, the modulator is a nucleic acid
of the invention. In other embodiments, the modulator is a peptide,
peptidomimetic, or other small molecule. In yet another embodiment,
the modulator is an antibody.
[0041] The present invention also provides diagnostic assays for
identifying the presence or absence of a genetic lesion or mutation
characterized by at least one of: (i) aberrant modification or
mutation of a gene encoding a polypeptide of the invention, (ii)
mis-regulation of a gene encoding a polypeptide of the invention,
and (iii) aberrant post-translational modification of a polypeptide
of the invention wherein a wild-type form of the gene encodes a
polypeptide having the activity of the polypeptide of the
invention.
[0042] In another aspect, the invention provides a method for
identifying a compound that binds with or modulates the activity of
a polypeptide of the invention. In general, such methods entail
measuring a biological activity of the polypeptide in the presence
and absence of a test compound and identifying those compounds
which bind with or alter the activity of the polypeptide.
[0043] The invention also features methods for identifying a
compound which modulates the expression of a polypeptide or nucleic
acid of the invention by measuring the expression of the
polypeptide or nucleic acid in the presence and absence of the
compound.
[0044] Other features and advantages of the invention will be
apparent from the following detailed description and claims.
BRIEF DESCRIPTION OF THE DRAWINGS
[0045] FIG. 1 comprises FIGS. 1A through 1J. The nucleotide
sequence (SEQ ID NO: 1) of a cDNA encoding the human TANGO 273
protein described herein is listed in FIGS. 1A-1C. The ORF
(residues 135 to 650; SEQ ID NO: 2) of the cDNA is indicated by
nucleotide triplets, above which the amino acid sequence (SEQ ID
NO: 3) of human TANGO 273 is listed. The nucleotide sequence (SEQ
ID NO: 11) of a cDNA encoding the murine TANGO 273 protein
described herein is listed in FIGS. 1D-1G. The ORF (residues 137 to
652; SEQ ID NO: 12) of the cDNA is indicated by nucleotide
triplets, above which the amino acid sequence (SEQ ID NO: 13) of
murine TANGO 273 is listed. An alignment of the amino acid
sequences of human ("Hum."; SEQ ID NO: 3) and murine ("Mur."; SEQ
ID NO: 13) TANGO 273 protein is shown in FIG. 1H, wherein identical
amino acid residues are indicated by ":" and similar amino acid
residues are indicated by ".". FIG. 1I is a hydrophobicity plot of
human TANGO 273 protein, and FIG. 1J is a hydrophobicity plot of
murine TANGO 273 protein.
[0046] FIG. 2 comprises FIGS. 2A through 2M-18. The nucleotide
sequence (SEQ ID NO: 21) of a cDNA encoding the human TANGO 325
protein described herein is listed in FIGS. 2A through 2E. The ORF
(residues 135 to 2000; SEQ ID NO: 22) of the cDNA is indicated by
nucleotide triplets, above which the amino acid sequence (SEQ ID
NO: 23) of human TANGO 325 is listed. FIG. 2F is a hydrophobicity
plot of TANGO 325 protein. An alignment of the amino acid sequences
of TANGO 325 ("325"; SEQ ID NO: 23) and Slit-1 protein ("Slit"; SEQ
ID NO: 29) protein is shown in FIGS. 2G to 2L. In FIGS. 2M-1 to
2M-18, an alignment of the nucleotide sequences of the cDNA
encoding human TANGO 325 protein ("325"; SEQ ID NO: 23) and the
nucleotide sequence of the cDNA encoding Slit-1 protein ("Slit";
SEQ ID NO: 30) is shown. This alignment was made using the ALIGN
software {Myers and Miller (1989) CABIOS, ver. 2.0}; pam120.mat
scoring matrix; gap opening penalty=12, gap extension
penalty=4).
[0047] FIG. 3 comprises FIGS. 3A through 3K. The nucleotide
sequence (SEQ ID NO: 31) of a cDNA encoding the human TANGO 364
protein described herein is listed in FIGS. 3A through 3E. The ORF
(residues 235 to 1764; SEQ ID NO: 32) of the cDNA is indicated by
nucleotide triplets, above which the amino acid sequence (SEQ ID
NO: 33) of human TANGO 364 is listed. FIG. 3F is a hydrophobicity
plot of human TANGO 364 protein. The nucleotide sequence (SEQ ID
NO: 41) of an alternatively-spliced form of the cDNA encoding the
human TANGO 364 protein described herein is listed in FIGS. 3G
through 31. The ORF (residues 2 to 898; SEQ ID NO: 42) of the cDNA
is indicated by nucleotide triplets, above which the amino acid
sequence (SEQ ID NO: 43) of the protein encoded by the splice
variant is listed. FIGS. 3J and 3K are an alignment of the amino
acid sequence of SEQ ID NOs: 33 and 43.
[0048] FIG. 4 comprises FIGS. 4A through 4P. The nucleotide
sequence (SEQ ID NO: 51) of a cDNA encoding the human TANGO 405
protein described herein is listed in FIGS. 4A through 4C. The ORF
(residues 154 to 780; SEQ ID NO: 52) of the cDNA is indicated by
nucleotide triplets, above which the amino acid sequence (SEQ ID
NO: 53) of human TANGO 405 is listed. FIG. 4D is a hydrophobicity
plot of human TANGO 405 protein. The nucleotide sequence (SEQ ID
NO: 61) of a cDNA encoding the murine TANGO 405 protein described
herein is listed in FIGS. 4E and 4F. The ORF (residues 174 to 707;
SEQ ID NO: 62) of the cDNA is indicated by nucleotide triplets,
above which the amino acid sequence (SEQ ID NO: 63) of murine TANGO
405 is listed. FIG. 4G is a hydrophobicity plot of murine TANGO 405
protein. An alignment of the amino acid sequences of human TANGO
405 protein (SEQ ID NO: 53) and murine TANGO 405 protein (SEQ ID
NO: 63) amino acid sequences is shown in FIG. 4H. An alignment of
the nucleotide sequences of the human (SEQ ID NO: 52) and murine
(SEQ ID NO: 62) ORFs encoding TANGO 405 protein is shown in FIGS.
4I through 4K. FIG. 4L is an alignment of the amino acid sequences
of murine TANGO 405 protein ("mT405"; SEQ ID NO: 63) and murine
dectin-2 ("Dectin"; SEQ ID NO: 60). FIG. 4M is an alignment of the
amino acid sequences of human TANGO 405 protein ("hT405"; SEQ ID
NO: 53) and murine dectin-2 ("Dectin"; SEQ ID NO: 60). The
nucleotide sequence (SEQ ID NO: 71) of an alternative embodiment of
a cDNA encoding the murine TANGO 405 protein described herein is
listed in FIGS. 4N, 4O and 4P. The ORF (residues 179 to 805; SEQ ID
NO: 72) of the cDNA is indicated by nucleotide triplets, above
which the amino acid sequence (SEQ ID NO: 73) of the alternative
embodiment of murine TANGO 405 is listed.
[0049] FIG. 5 comprises FIGS. 5A through 5C. The nucleotide
sequence (SEQ ID NO: 81) of a cDNA encoding the human M019 (i.e.,
TANGO 533) protein described herein is listed in FIGS. 5A and 5B.
The ORF (residues 331 to 585; SEQ ID NO: 82) of the cDNA is
indicated by nucleotide triplets, above which the amino acid
sequence (SEQ ID NO: 83) of human M019 protein is listed. FIG. 5C
is a hydrophobicity plot of human M019 protein, in which the
locations of cysteine residues ("Cys"), and the predicted
extracellular ("out"), intracellular ("ins"), or transmembrane
("TM") locations of the protein backbone is indicated by a
horizontal bar.
DETAILED DESCRIPTION OF THE INVENTION
[0050] The present invention is based, at least in part, on the
discovery of a variety of cDNA molecules which encode proteins
which are herein designated TANGO 273, TANGO 325, TANGO 364, TANGO
405, and M019. These proteins exhibit a variety of physiological
activities, and are included in a single application for the sake
of convenience. It is understood that the allowability or
non-allowability of claims directed to one of these proteins has no
bearing on the allowability of claims directed to the others. The
characteristics of each of these proteins and the cDNAs encoding
them are described separately in the ensuing sections. In addition
to the full length mature and immature proteins described in the
following sections, the invention includes fragments, derivatives,
and variants of these proteins, as described herein. These
proteins, fragments, derivatives, and variants are collectively
referred to herein as polypeptides of the invention or proteins of
the invention.
TANGO 273
[0051] A cDNA clone (designated jthoc028g06) encoding at least a
portion of human TANGO 273 protein was isolated from a
lipopolysaccharide-(LPS)stimulated human osteoblast cDNA library.
The corresponding murine cDNA clone (designated jtmoa001c04) was
isolated from an LPS-stimulated murine osteoblast cDNA library. The
human and murine TANGO 273 proteins are predicted by structural
analysis to be transmembrane proteins.
[0052] The full length of the cDNA encoding human TANGO 273 protein
(FIG. 1; SEQ ID NO: 1) is 2964 nucleotide residues. The ORF of this
cDNA, nucleotide residues 135 to 650 of SEQ ID NO: 1 (i.e., SEQ ID
NO: 2), encodes a 172-amino acid transmembrane protein (FIG. 1; SEQ
ID NO: 3).
[0053] The invention thus includes purified human TANGO 273
protein, both in the form of the immature 172 amino acid residue
protein (SEQ ID NO: 3) and in the form of the mature 150 amino acid
residue protein (SEQ ID NO: 5). The invention also includes
purified murine TANGO 273 protein, both in the form of the immature
172 amino acid residue protein (SEQ ID NO: 13) and in the form of
the mature 150 amino acid residue protein (SEQ ID NO: 15). Mature
human or murine TANGO 273 proteins can be synthesized without the
signal sequence polypeptide at the amino terminus thereof, or they
can be synthesized by generating immature TANGO 273 protein and
cleaving the signal sequence therefrom.
[0054] The invention also includes nucleic acid molecules which
encode a polypeptide of the invention. Such nucleic acids include,
for example, a DNA molecule having the nucleotide sequence listed
in SEQ ID NO: 1 or some portion thereof or SEQ ID NO: 12 or some
portion thereof, such as the portion which encodes mature TANGO 273
protein, immature TANGO 273 protein, or a domain of TANGO 273
protein. These nucleic acids are collectively referred to as
nucleic acids of the invention.
[0055] TANGO 273 proteins and nucleic acid molecules encoding them
comprise a family of molecules having certain conserved structural
and functional features. This family includes, by way of example,
the human and murine TANGO 273 proteins.
[0056] A common domain of TANGO 273 proteins is a signal sequence.
As used herein, a signal sequence includes a peptide of at least
about 10 amino acid residues in length which occurs at the amino
terminus of membrane-bound proteins and which contains at least
about 45% hydrophobic amino acid residues such as alanine, leucine,
isoleucine, phenylalanine, proline, tyrosine, tryptophan, or
valine. In a preferred embodiment, a signal sequence contains at
least about 10 to 35 amino acid residues, preferably about 10 to 20
amino acid residues, and has at least about 35-60%, more preferably
40-50%, and more preferably at least about 45% hydrophobic
residues. A signal sequence serves to direct a protein containing
such a sequence to a lipid bilayer. Thus, in one embodiment, a
TANGO 273 protein contains a signal sequence corresponding to amino
acid residues 1 to 22 of SEQ ID NO: 3 (SEQ ID NO: 4) or to amino
acid residues 1 to 22 of SEQ ID NO: 13. The signal sequence is
cleaved during processing of the mature protein.
[0057] TANGO 273 proteins can also include an extracellular domain.
The human TANGO 273 protein extracellular domain is located from
about amino acid residue 23 to about amino acid residue 60 of SEQ
ID NO: 3, and the murine TANGO 273 protein extracellular domain is
located from about amino acid residue 23 to about amino acid
residue 60 of SEQ ID NO: 13.
[0058] The present invention also includes TANGO 273 proteins
having a transmembrane domain. As used herein, a "transmembrane
domain" refers to an amino acid sequence having at least about 15
to 50 amino acid residues in length and which contains at least
about 65-70% hydrophobic amino acid residues such as alanine,
leucine, phenylalanine, protein, tyrosine, tryptophan, or valine.
In a preferred embodiment, a transmembrane domain has at least
about 60-80%, more preferably 65-75%, and more preferably at least
about 70% hydrophobic residues. Thus, in one embodiment, a human
TANGO 273 protein of the invention contains a transmembrane domain
corresponding to about amino acid residues 61 to 81 of SEQ ID NO: 3
(SEQ ID NO: 7). In another embodiment, a murine TANGO 273 protein
of the invention contains a transmembrane domain corresponding to
about amino acid residues 61 to 81 of SEQ ID NO: 13.
[0059] In addition, TANGO 273 proteins include a cytoplasmic
domain. The human TANGO 273 cytoplasmic domain is located from
about amino acid residue 82 to amino acid residue 172 of SEQ ID NO:
3 (SEQ ID NO: 8), and the murine TANGO 273 cytoplasmic domain is
located from about amino acid residue 82 to amino acid residue 172
of SEQ ID NO: 13.
[0060] TANGO 273 proteins typically comprise a variety of potential
post-translational modification sites (often within an
extracellular domain), such as those described herein in Tables I
and II, as predicted by computerized sequence analysis of human and
murine TANGO 273 proteins using amino acid sequence comparison
software (comparing the amino acid sequence of TANGO 273 with the
information in the PROSITE database {rel. 12.2; February, 1995} and
the Hidden Markov Models database {Rel. PFAM 3.3}). As used herein,
the term "post-translational modification site" refers to a protein
domain that includes about 3 to 10 amino acid residues, more
preferably about 3 to 6 amino acid residues wherein the domain has
an amino acid sequence which comprises a consensus sequence which
is recognized and modified by a protein-modifying enzyme. Examples
of protein-modifying enzymes include amino acid glycosylases, cAMP-
and cGMP-dependent protein kinases, protein kinase C, casein kinase
II, and myristoylases. In certain embodiments, a protein of the
invention has at least 1, 2, 3, 4, 5, or all 6 of the
post-translational modification sites listed in Table I. In other
embodiments, the protein of the invention has at least 1, 2, 3, 4,
5, 6, or all 7 of the post-translational modification sites listed
in Table II.
TABLE-US-00001 TABLE I Amino Acid Amino Type of Potential Residues
of Acid Modification Site or Domain SEQ ID NO: 3 Sequence
N-glycosylation site 97 to 100 NVSY Casein kinase II phosphory- 41
to 44 SYED lation site N-myristoylation site 31 to 36 GLYPTY 47 to
52 GSRCCV 70 to 75 GVLFCC 131 to 136 GNSMAM Src Homology 3 (SH3)
domain 86 to 90 YPPPL binding site 103 to 107 QPPNP 113 to 117
QPGPP 121 to 125 DPGGP 140 to 145 VPPNSP 151 to 155 CPPPP 160 to
164 TPPPP
TABLE-US-00002 TABLE II Amino Acid Amino Type of Potential Residues
of Acid Modification Site or Domain SEQ ID NO: 13 Sequence
N-glycosylation site 97 to 100 NVSY Casein kinase II phosphory- 41
to 44 SYED lation site N-myristoylation site 31 to 36 GLYPTY 47 to
52 GSRCCV 70 to 75 GVLFCC 131 to 136 GNTMAM Src Homology 3 (SH3)
domain 86 to 90 YPPPL binding site 103 to 107 QPPNP 115 to 119
GPPYY 121 to 125 DPGGP 141 to 145 QPNSP 151 to 155 YPPPP 160 to 164
TPPPP Amidation site 1 to 4 MGRR
[0061] The amino acid sequence of TANGO 273 protein includes about
seven potential proline-rich Src homology 3 (SH3) domain binding
sites nearer the cytoplasmic portion of the protein. SH3 domains
mediate specific assembly of protein complexes, presumably by
interacting with proline-rich protein domains (Morton and Campbell
(1994) Curr. Biol. 4:615-617). SH3 domains also mediate
interactions between proteins involved in transmembrane signal
transduction. Coupling of proteins mediated by SH3 domains has been
implicated in a variety of physiological systems, including those
involving regulation of cell growth and proliferation, endocytosis,
and activation of respiratory burst.
[0062] SH3 domains have been described in the art (e.g., Mayer et
al. (1988) Nature 332:272-275; Musacchio et al. (1992) FEBS Lett.
307:55-61; Pawson and Schlessinger (1993) Curr. Biol. 3:434-442;
Mayer and Baltimore (1993) Trends Cell Biol. 3:8-13; Pawson (1993)
Nature 373:573-580), and occur in a variety of cytoplasmic
proteins, including several (e.g., protein tyrosine kinases)
involved in transmembrane signal transduction. Among the proteins
in which one or more SH3 domains occur are protein tyrosine kinases
such as those of the Src, Abl, Bkt, Csk and ZAP70 families,
mammalian phosphatidylinositol-specific phospholipases C-gamma-1
and -2, mammalian phosphatidylinositol 3-kinase regulatory p85
subunit, mammalian Ras GTPase-activating protein (GAP), proteins
which mediate binding of guanine nucleotide exchange factors and
growth factor receptors (e.g., vertebrate GRB2, Caenorhabditis
elegans sem-5, and Drosophila DRK proteins), mammalian Vav
oncoprotein, guanidine nucleotide releasing factors of the CDC 25
family (e.g., yeast CDC25, yeast SCD25, and fission yeast ste6
proteins), MAGUK proteins (e.g., mammalian tight junction protein
ZO-1, vertebrate erythrocyte membrane protein p55, C. elegans
protein lin-2, rat protein CASK, and mammalian synaptic proteins
SAP90/PSD-95, CHAPSYN-110/PSD-93, SAP97/DLG 1, and SAP 102),
proteins which interact with vertebrate receptor protein tyrosine
kinases (e.g., mammalian cytoplasmic protein Nck and oncoprotein
Crk), chicken Src substrate p80/85 protein (cortactin), human
hemopoietic lineage cell specific protein Hsl, mammalian
dihydrouridine-sensitive L-type calcium channel beta subunit, human
myasthenic syndrome antigen B (MSYB), mammalian neutrophil
cytosolic activators of NADPH oxidase (e.g., p47 {NCF-1}, p67
{NCF-2}, and C. elegans protein B0303.7) myosin heavy chains (MYO3)
from amoebae, from slime molds, and from yeast, vertebrate and
Drosophila spectrin and fodrin alpha chain proteins, human
amphiphysin, yeast actin-binding proteins ABP1 and SLA3, yeast
protein BEM1, fission yeast protein scd2 (ral3), yeast BEM1-binding
proteins B012 (BEB1) and BOB1 (BOI1), yeast fusion protein FUS1,
yeast protein RSV167, yeast protein SSU81, yeast hypothetical
proteins YAR014c, YFR024c, YHL002w, YHR016c, YJL020C, and YHR114w,
hypothetical fission yeast protein SpAC12C2.05c, and C. elegans
hypothetical protein F42H10.3. Of these proteins, multiple SH3
domains occur in vertebrate GRB2 protein, C. elegans sem-5 protein,
Drosophila DRK protein, oncoprotein Crk, mammalian neutrophil
cytosolic activators of NADPH oxidase p47 and p67, yeast protein
BEM1, fission yeast protein scd2, yeast hypothetical protein
YHR114w, mammalian cytoplasmic protein Nck, C. elegans neutrophil
cytosolic activator of NADPH oxidase B0303.7, and yeast
actin-binding protein SLA1. Of these proteins, three or more SH3
domains occur in mammalian cytoplasmic protein Nck, C. elegans
neutrophil cytosolic activator of NADPH oxidase B0303.7, and yeast
actin-binding protein SLA1. The presence of SH3 domain binding
sites in TANGO 273 indicates that TANGO 273 interacts with one or
more of these and other SH3 domain-containing proteins and is thus
involved in physiological processes in which one or more of these
or other SH3 domain-containing proteins are involved.
[0063] The signal peptide prediction program SIGNALP (Nielsen et
al. (1997) Protein Engineering 10:1-6) predicted that human TANGO
273 protein includes a 22 amino acid signal peptide (amino acid
residues 1 to 22 of SEQ ID NO: 3; SEQ ID NO: 4) preceding the
mature TANGO 273 protein (amino acid residues 23 to 172 of SEQ ID
NO: 3; SEQ ID NO: 5). Human TANGO 273 protein includes an
extracellular domain (amino acid residues 23 to 60 of SEQ ID NO: 3;
SEQ ID NO: 6); a transmembrane domain (amino acid residues 61 to 81
of SEQ ID NO: 3; SEQ ID NO: 7); and a cytoplasmic domain (amino
acid residues 82 to 172 of SEQ ID NO: 3; SEQ ID NO: 8).
[0064] FIG. 1I depicts a hydrophobicity plot of human TANGO 273
protein. Relatively hydrophobic regions are above the dashed
horizontal line, and relatively hydrophilic regions are below the
dashed horizontal line. The hydrophobic region which corresponds to
amino acid residues 1 to 22 of SEQ ID NO: 3 is the signal sequence
of human TANGO 273 (SEQ ID NO: 4). The hydrophobic region which
corresponds to amino acid residues 61 to 81 of SEQ ID NO: 3 is the
transmembrane domain of human TANGO 273 (SEQ ID NO: 7). As
described elsewhere herein, relatively hydrophilic regions are
generally located at or near the surface of a protein, and are more
frequently effective immunogenic epitopes than are relatively
hydrophobic regions. For example, the region of human TANGO 273
protein from about amino acid residue 100 to about amino acid
residue 120 appears to be located at or near the surface of the
protein, while the region from about amino acid residue 130 to
about amino acid residue 140 appears not to be located at or near
the surface.
[0065] Chromosomal mapping was performed by computerized comparison
of TANGO 273 cDNA sequences against a chromosomal mapping database
in order to identify the approximate location of the gene encoding
human TANGO 273 protein. This analysis indicated that the gene is
located on chromosome 7 between markers D7S2467 and D7S2552.
[0066] The predicted molecular weight of human TANGO 273 protein
without modification and prior to cleavage of the signal sequence
is about 19.2 kilodaltons. The predicted molecular weight of the
mature human TANGO 273 protein without modification and after
cleavage of the signal sequence is about 16.8 kilodaltons.
[0067] Northern analysis experiments indicated that mRNA
corresponding to the cDNA encoding TANGO 273 is expressed in the
tissues listed in Table III, wherein "++" indicates moderate
expression and "+" indicates lower expression.
TABLE-US-00003 TABLE III Animal Tissue Relative Level of Expression
Human heart ++ brain ++ skeletal muscle ++ pancreas ++ placenta +
lung + liver + kidney +
[0068] The full length of the cDNA encoding murine TANGO 273
protein (FIG. 1; SEQ ID NO: 11) is 2915 nucleotide residues. The
ORF of this cDNA, nucleotide residues 137 to 650 of SEQ ID NO: 11
(i.e., SEQ ID NO: 12), encodes a 172-amino acid transmembrane
protein (FIG. 1; SEQ ID NO: 13).
[0069] The signal peptide prediction program SIGNALP (Nielsen et
al. (1997) Protein Engineering 10:1-6) predicted that murine TANGO
273 protein includes a 22 amino acid signal peptide (amino acid
residues 1 to 22 of SEQ ID NO: 13) preceding the mature TANGO 273
protein (amino acid residues 23 to 172 of SEQ ID NO: 13; SEQ ID NO:
15). Murine TANGO 273 protein includes an extracellular domain
(amino acid residues 23 to 60 of SEQ ID NO: 13); a transmembrane
domain (amino acid residues 61 to 81 of SEQ ID NO: 13); and a
cytoplasmic domain (amino acid residues 82 to 172 of SEQ ID NO:
13).
[0070] FIG. 1J depicts a hydrophobicity plot of murine TANGO 273
protein. Relatively hydrophobic regions are above the dashed
horizontal line, and relatively hydrophilic regions are below the
dashed horizontal line. The hydrophobic region which corresponds to
amino acid residues 1 to 22 of SEQ ID NO: 13 is the signal sequence
of murine TANGO 273. As described elsewhere herein, relatively
hydrophilic regions are generally located at or near the surface of
a protein, and are more frequently effective immunogenic epitopes
than are relatively hydrophobic regions. For example, the region of
murine TANGO 273 protein from about amino acid residue 100 to about
amino acid residue 120 appears to be located at or near the surface
of the protein, while the region from about amino acid residue 130
to about amino acid residue 140 appears not to be located at or
near the surface.
[0071] The predicted molecular weight of murine TANGO 273 protein
without modification and prior to cleavage of the signal sequence
is about 19.4 kilodaltons. The predicted molecular weight of the
mature murine TANGO 273 protein without modification and after
cleavage of the signal sequence is about 17.1 kilodaltons.
[0072] In situ analysis of murine TANGO 273 mRNA indicated that
TANGO 273 is expressed in central nervous system (CNS) tissues
during embryogenesis and into adulthood. Expression of TANGO 273 is
widely observed in murine CNS tissues, including brain, spinal
cord, eye, and olfactory epithelium at all embryonic ages examined
(i.e., at embryonic days 13.5, 14.5, 15.5, 16.5, and 18.5 and at
post-natal day 1.5).
[0073] Human and murine TANGO 273 cDNA sequences exhibit
significant nucleotide sequence identity with an expressed sequence
tag (EST) isolated from a library of ESTs corresponding to proteins
secreted from prostate tissue, as described in PCT publication
number WO 99/06550, published Feb. 11, 1999.
[0074] Human and murine TANGO 273 proteins exhibit considerable
sequence similarity, as indicated herein in FIG. 1H. FIG. 1H
depicts an alignment of human and murine TANGO 273 protein amino
acid sequences (SEQ ID NOs: 3 and 13, respectively). In this
alignment (pam120.mat scoring matrix, gap penalties -12/-4), the
proteins are 89.5% identical. Alignment of the ORF encoding human
TANGO 273 protein and the ORF encoding murine TANGO 273 protein
using the same software and parameters indicated that the
nucleotide sequences are 84.1% identical.
[0075] Uses of TANGO 273 Nucleic Acids,
[0076] Polypeptides, and Modulators Thereof
[0077] cDNAs encoding the human and murine TANGO 273 proteins were
each isolated from LPS-stimulated osteoblast cDNA libraries. These
proteins are involved in bone-related metabolism, homeostasis, and
development disorders. Thus, proteins and nucleic acids of the
invention which are identical to, similar to, or derived from human
and murine TANGO 273 proteins and nucleic acids encoding them are
useful for preventing, diagnosing, and treating, among others,
bone-related disorders such as osteoporosis, cancer, skeletal
development disorders, bone fragility, and the like.
[0078] Expression of TANGO 273 in heart, brain, skeletal muscle,
and pancreas, placenta, lung, liver, and kidney tissues is an
indication that TANGO 273 proteins, nucleic acids encoding them,
and agents that modulate activity or expression of either of these
can be used to modulate growth, proliferation, survival,
differentiation, adhesion, and activity of cells of these tissues,
or to prognosticate, diagnose, and treat one or more disorders
which affect these tissues.
[0079] The fact that TANGO 273 is expressed at high levels in
neurological tissues is an indication that TANGO 273 proteins,
nucleic acids, and modulators thereof can be used to modulate
proliferation, differentiation, or function of neurological cells
in these tissues (e.g., neuronal cells). Thus, TANGO 273 proteins,
nucleic acids, and modulators thereof can be used to prognosticate,
diagnose, and treat one or more neurological disorders. Examples of
such disorders include CNS disorders, CNS-related disorders, focal
brain disorders, global-diffuse cerebral disorders, and other
neurological and cerebrovascular disorders.
[0080] CNS disorders include, but are not limited to cognitive and
neurodegenerative disorders such as Alzheimer's disease, senile
dementia, Huntington's disease, amyotrophic lateral sclerosis, and
Parkinson's disease, as well as Gilles de la Tourette's syndrome,
autonomic function disorders such as hypertension and sleep
disorders (e.g., insomnia, hypersomnia, parasomnia, and sleep
apnea); neuropsychiatric disorders (e.g., schizophrenia,
schizoaffective disorder, attention deficit disorder, dysthymic
disorder, major depressive disorder, mania, and
obsessive-compulsive disorder); psychoactive substance use
disorders; anxiety; panic disorder; and bipolar affective disorders
(e.g., severe bipolar affective disorder and bipolar affective
disorder with hypomania and major depression).
[0081] CNS-related disorders include disorders associated with
developmental, cognitive, and autonomic neural and neurological
processes, such as pain, appetite, long term memory, and short term
memory.
[0082] Examples of focal brain disorders include aphasia, apraxia,
agnosia, and amnesias (e.g., posttraumatic amnesia, transient
global amnesia, and psychogenic amnesia). Global-diffuse cerebral
disorders with which TANGO 273 can be associated include coma,
stupor, obtundation, and disorders of the reticular formation.
[0083] Other neurological disorders with which TANGO 273 can be
associated include ischemic syndromes (e.g., stroke), hypertensive
encephalopathy, hemorrhagic disorders, and disorders involving
aberrant function of the blood-brain barrier (e.g., CNS infections
such as meningitis and encephalitis, aseptic meningitis, metastasis
of non-CNS tumor cells into the CNS, various pain disorders such as
migraine, blindness and other vision problems, and CNS-related
adverse drug reactions such as head pain, sleepiness, and
confusion). TANGO 273 proteins, nucleic acids encoding them, and
agents that modulate activity or expression of either of these can
be used to prognosticate, diagnose, and treat one or more of these
disorders.
[0084] Developmental regulation of TANGO 273 expression in fetal
neurological tissues, as described herein, is an indication that
TANGO 273 proteins, nucleic acids, and modulators thereof can be
used to prognosticate, diagnose, and treat one or more disorders
which involve aberrant fetal neurological development. Examples of
such disorders include blindness, deafness, fetal death, mental
retardation, dysraphia, anencephaly, malformation of cerebral
hemispheres, encephalocele, porencephaly, hydranencephaly,
hydrocephalus, and spina bifida.
[0085] The fact that TANGO 273 is expressed in tissues which were
exposed to LPS indicates that TANGO 273 mediates one or more
physiological responses of cells to bacterial infection. Thus,
TANGO 273 is involved in one or more of detection of bacteria in a
tissue in which it is expressed, movement of cells with relation to
sites of bacterial infection, production of biological molecules
which inhibit bacterial infection, and production of biological
molecules which alleviate cellular or other physiological damage
wrought by bacterial infection.
[0086] Presence in TANGO 273 protein of multiple SH3 domain binding
sites indicates that TANGO 273 protein interacts with one or more
SH3 domain-containing proteins. Thus, TANGO 273 protein mediates
binding of proteins (i.e., binding of proteins to TANGO 273 and to
one another to form protein complexes) in cells in which it is
expressed. TANGO 273 is also involved in transduction of signals
between the exterior environment of cells (i.e., including from
other cells) and the interior of cells in which it is expressed.
TANGO 273 mediates regulation of cell growth and proliferation,
endocytosis, activation of respiratory burst, and other
physiological processes triggered by transmission of a signal via a
protein with which TANGO 273 interacts.
[0087] Sequence similarity of TANGO 273 cDNA with an EST expressed
in prostate tissue indicates that TANGO 273 can be expressed in
prostate tissue, and can thus be involved in disorders of the
prostate. Thus, TANGO 273 proteins, nucleic acids encoding them,
and agents that modulate activity or expression of either of these
can be used to treat prostate disorders. Examples of prostate
disorders which can be treated in this manner include inflammatory
prostatic diseases (e.g., acute and chronic prostatitis and
granulomatous prostatitis), prostatic hyperplasia (e.g., benign
prostatic hypertrophy or hyperplasia), and prostate tumors (e.g.,
carcinomas).
[0088] In another example, TANGO 273 polypeptides, nucleic acids,
or modulators thereof, can be used to treat cardiovascular
disorders. Examples of cardiac disorders which can be treated in
this manner include ischemic heart diseases (e.g., angina pectoris,
myocardial infarction and its aftermath, coronary artery disease,
cardiac arrest, and chronic ischemic heart disease), hypertensive
heart disease, pulmonary heart disease, valvular heart disease
(e.g., rheumatic fever and rheumatic heart disease, endocarditis,
mitral valve prolapse, and aortic valve stenosis), congenital heart
disease (e.g., valvular and vascular obstructive lesions, atrial or
ventricular septal defect, and patent ductus arteriosus), cardiac
arrhythmia, cardiac insufficiency, endocarditis, pericardial
disease, muscular dystrophy, and myocardial disease (e.g.,
myocarditis, congestive cardiomyopathy, restrictive cardiomyopathy,
and hypertrophic cardiomyopathy). Examples of vascular disorders
which can be treated in this manner include arteriosclerosis,
atherosclerosis, hypertension, aberrant or non-desired
angiogenesis, stenosis and restenosis, and smooth muscle
proliferation in response to traumatic injury.
[0089] In another example, TANGO 273 polypeptides, nucleic acids,
or modulators thereof, can be used to treat disorders of the brain.
Examples of brain disorders in which TANGO 273 can have role
include both CNS disorders, CNS-related disorders, focal brain
disorders, global-diffuse cerebral disorders, and other
neurological and cerebrovascular disorders. CNS disorders include,
but are not limited to cognitive and neurodegenerative disorders
such as Alzheimer's disease, senile dementia, Huntington's disease,
amyotrophic lateral sclerosis, and Parkinson's disease, as well as
Gilles de la Tourette's syndrome, autonomic function disorders such
as hypertension and sleep disorders (e.g., insomnia, hypersomnia,
parasomnia, and sleep apnea), neuropsychiatric disorders (e.g.,
schizophrenia, schizoaffective disorder, attention deficit
disorder, dysthymic disorder, major depressive disorder, mania, and
obsessive-compulsive disorder), psychoactive substance use
disorders, anxiety, panic disorder, and bipolar affective disorder
(e.g., severe bipolar affective disorder and bipolar affective
disorder with hypomania and major depression). CNS-related
disorders include disorders associated with developmental,
cognitive, and autonomic neural and neurological processes, such as
pain, appetite, long term memory, and short term memory. Examples
of focal brain disorders include aphasia, apraxia, agnosia, and
amnesias (e.g., posttraumatic amnesia, transient global amnesia,
and psychogenic amnesia). Global-diffuse cerebral disorders with
which TANGO 273 is associated include coma, stupor, obtundation,
and disorders of the reticular formation. Cerebrovascular disorders
include ischemic syndromes (e.g., stroke), hypertensive
encephalopathy, hemorrhagic disorders, and disorders involving
aberrant function of the blood-brain barrier (e.g., CNS infections
such as meningitis and encephalitis, aseptic meningitis, metastasis
of non-CNS tumor cells into the CNS, various pain disorders such as
migraine, and CNS-related adverse drug reactions such as head pain,
sleepiness, and confusion).
[0090] In another example, TANGO 273 polypeptides, nucleic acids,
or modulators thereof, can be used to treat disorders of skeletal
muscle, such as muscular dystrophy (e.g., Duchenne muscular
dystrophy, Becker muscular dystrophy, Emery-Dreifuss muscular
dystrophy, limb-girdle muscular dystrophy, facioscapulohumeral
muscular dystrophy, myotonic dystrophy, oculopharyngeal muscular
dystrophy, distal muscular dystrophy, and congenital muscular
dystrophy), motor neuron diseases (e.g., amyotrophic lateral
sclerosis, infantile progressive spinal muscular atrophy,
intermediate spinal muscular atrophy, spinal bulbar muscular
atrophy, and adult spinal muscular atrophy), myopathies (e.g.,
inflammatory myopathies such as dermatomyositis and polymyositis,
myotonia congenita, paramyotonia congenita, central core disease,
nemaline myopathy, myotubular myopathy, and periodic paralysis),
and metabolic diseases of muscle (e.g., phosphorylase deficiency,
acid maltase deficiency, phosphofructokinase deficiency, debrancher
enzyme deficiency, mitochondrial myopathy, carnitine deficiency,
carnitine palmityl transferase deficiency, phosphoglycerate kinase
deficiency, phosphoglycerate mutase deficiency, lactate
dehydrogenase deficiency, and myoadenylate deaminase
deficiency).
[0091] In another example, TANGO 273 polypeptides, nucleic acids,
or modulators thereof, can be used to treat pancreatic disorders,
such as pancreatitis (e.g., acute hemorrhagic pancreatitis and
chronic pancreatitis), pancreatic cysts (e.g., congenital cysts,
pseudocysts, and benign or malignant neoplastic cysts), pancreatic
tumors (e.g., pancreatic carcinoma and adenoma), diabetes mellitus
(e.g., insulin- and non-insulin-dependent types, impaired glucose
tolerance, and gestational diabetes), or islet cell tumors (e.g.,
insulinomas, adenomas, Zollinger-Ellison syndrome, glucagonomas,
and somatostatinoma).
[0092] In another example, TANGO 273 polypeptides, nucleic acids,
or modulators thereof, can be used to treat placental disorders,
such as toxemia of pregnancy (e.g., preeclampsia and eclampsia),
placentitis, or spontaneous abortion.
[0093] In another example, TANGO 273 polypeptides, nucleic acids,
or modulators thereof, can be used to treat pulmonary disorders,
such as atelectasis, cystic fibrosis, rheumatoid lung disease,
pulmonary congestion or edema, chronic obstructive airway disease
(e.g., emphysema, chronic bronchitis, bronchial asthma, and
bronchiectasis), diffuse interstitial diseases (e.g., sarcoidosis,
pneumoconiosis, hypersensitivity pneumonitis, Goodpasture's
syndrome, idiopathic pulmonary hemosiderosis, pulmonary alveolar
proteinosis, desquamative interstitial pneumonitis, chronic
interstitial pneumonia, fibrosing alveolitis, hamman-rich syndrome,
pulmonary eosinophilia, diffuse interstitial fibrosis, Wegener's
granulomatosis, lymphomatoid granulomatosis, and lipid pneumonia),
or tumors (e.g., bronchogenic carcinoma, bronchioalveolar
carcinoma, bronchial carcinoid, hamartoma, and mesenchymal
tumors).
[0094] In another example, TANGO 273 polypeptides, nucleic acids,
or modulators thereof, can be used to treat hepatic (liver)
disorders, such as jaundice, hepatic failure, hereditary
hyperbilirubinemias (e.g., Gilbert's syndrome, Crigler-Naijar
syndromes, and Dubin-Johnson and Rotor's syndromes), hepatic
circulatory disorders (e.g., hepatic vein thrombosis and portal
vein obstruction and thrombosis) hepatitis (e.g., chronic active
hepatitis, acute viral hepatitis, and toxic and drug-induced
hepatitis) cirrhosis (e.g., alcoholic cirrhosis, biliary cirrhosis,
and hemochromatosis), or malignant tumors (e.g., primary carcinoma,
hepatoblastoma, and angiosarcoma).
[0095] In another example, TANGO 273 polypeptides, nucleic acids,
or modulators thereof, can be used to treat renal (kidney)
disorders, such as glomerular diseases (e.g., acute and chronic
glomerulonephritis, rapidly progressive glomerulonephritis,
nephrotic syndrome, focal proliferative glomerulonephritis,
glomerular lesions associated with systemic disease such as
systemic lupus erythematosus, Goodpasture's syndrome, multiple
myeloma, diabetes, neoplasia, sickle cell disease, and chronic
inflammatory diseases), tubular diseases (e.g., acute tubular
necrosis and acute renal failure, polycystic renal disease,
medullary sponge kidney, medullary cystic disease, nephrogenic
diabetes, and renal tubular acidosis), tubulointerstitial diseases
(e.g., pyelonephritis, drug and toxin induced tubulointerstitial
nephritis, hypercalcemic nephropathy, and hypokalemic nephropathy)
acute and rapidly progressive renal failure, chronic renal failure,
nephrolithiasis, vascular diseases (e.g., hypertension and
nephrosclerosis, microangiopathic hemolytic anemia, atheroembolic
renal disease, diffuse cortical necrosis, and renal infarcts), or
tumors (e.g., renal cell carcinoma and nephroblastoma).
TANGO 325
[0096] A cDNA clone (designated jthdc071a12) encoding at least a
portion of human TANGO 325 protein was isolated from a human aortic
endothelial cell cDNA library. The human TANGO 325 protein is
predicted by structural analysis to be a transmembrane protein.
[0097] The full length of the cDNA encoding human TANGO 325 protein
(FIG. 2; SEQ ID NO: 21) is 2169 nucleotide residues. The ORF of
this cDNA, nucleotide residues 135 to 2000 of SEQ ID NO: 21 (i.e.,
SEQ ID NO: 22), encodes a 622-amino acid transmembrane protein
(FIG. 2; SEQ ID NO: 23).
[0098] The invention thus includes purified human TANGO 325
protein, both in the form of the immature 622 amino acid residue
protein (SEQ ID NO: 23) and in the form of the mature,
approximately 591 amino acid residue protein (SEQ ID NO: 25).
Mature human TANGO 325 protein can be synthesized without the
signal sequence polypeptide at the amino terminus thereof, or it
can be synthesized by generating immature TANGO 325 protein and
cleaving the signal sequence therefrom.
[0099] The invention also includes nucleic acid molecules which
encode a TANGO 325 polypeptide of the invention. Such nucleic acids
include, for example, a DNA molecule having the nucleotide sequence
listed in SEQ ID NO: 21 or some portion thereof, such as the
portion which encodes mature TANGO 325 protein, immature TANGO 325
protein, or a domain of TANGO 325 protein. These nucleic acids are
collectively referred to as TANGO 325 nucleic acids of the
invention.
[0100] TANGO 325 proteins and nucleic acid molecules encoding them
comprise a family of molecules having certain conserved structural
and functional features.
[0101] A common domain present in TANGO 325 proteins is a signal
sequence. In one embodiment, a TANGO 325 protein contains a signal
sequence corresponding to about amino acid residues 1 to 31 of SEQ
ID NO: 23 (SEQ ID NO: 24). The signal sequence is cleaved during
processing of the mature protein.
[0102] TANGO 325 proteins can include an extracellular domain. The
human TANGO 325 protein extracellular domain is located from about
amino acid residue 32 to about amino acid residue 529 of SEQ ID NO:
23 (SEQ ID NO: 26).
[0103] In addition, TANGO 325 include a transmembrane domain. In
one embodiment, a TANGO 325 protein of the invention contains a
transmembrane domain corresponding to about amino acid residues 530
to 547 of SEQ ID NO: 23 (SEQ ID NO: 27).
[0104] The present invention includes TANGO 325 proteins having a
cytoplasmic domain, particularly including proteins having a
carboxyl-terminal cytoplasmic domain. The human TANGO 325
cytoplasmic domain is located from about amino acid residue 548 to
amino acid residue 622 of SEQ ID NO: 23 (SEQ ID NO: 28).
[0105] TANGO 325 proteins typically comprise a variety of potential
post-translational modification sites (often within an
extracellular domain), such as those described herein in Table IV,
as predicted by computerized sequence analysis of TANGO 325
proteins using amino acid sequence comparison software (comparing
the amino acid sequence of TANGO 325 with the information in the
PROSITE database {rel. 12.2; February, 1995} and the Hidden Markov
Models database {Rel. PFAM 3.3}). In certain embodiments, a protein
of the invention has at least 1, 2, 4, 6, 10, 15, or 20 or more of
the post-translational modification sites listed in Table IV.
TABLE-US-00004 TABLE IV Amino Acid Amino Type of Potential Residues
of Acid Modification Site or Domain SEQ ID NO: 23 Sequence
N-glycosylation site 71 to 74 NISY 76 to 79 NESE 215 to 218 NLTK
266 to 269 NVTR 317 to 320 NDTF 331 to 334 NLSF 336 to 339 NLTA 400
to 403 NITN 410 to 413 NVSR 451 to 454 NITF 579 to 582 NVTA cAMP-
or cGMP-dependent 231 to 234 RRLS protein kinase phosphory- lation
site Protein kinase C phosphory- 40 to 42 TGR lation site 229 to
231 SLR 326 to 328 SLK 390 to 392 SMR 510 to 512 SGK 575 to 577 SAR
Casein kinase II phosphory- 284 to 287 SHND lation site 442 to 445
SPLE 447 to 450 TETE 453 to 456 TFWE N-myristoylation site 3 to 8
GLQFSL 69 to 74 GNNISY 126 to 131 GIFKGL 174 to 179 GTFVGM
ATP/GTP-binding site motif A 506 to 513 AASMSGKT (P-loop) Leucine
rich repeat amino 32 to 60 See FIG. terminal domain (LLRNT) 2
Leucine rich repeat (LRR) 61 to 84 See FIG. domain 2 85 to 108 See
FIG. 2 109 to 132 See FIG. 2 133 to 156 See FIG. 2 157 to 180 See
FIG. 2 181 to 204 See FIG. 2 205 to 228 See FIG. 2 229 to 252 See
FIG. 2 253 to 276 See FIG. 2 277 to 300 See FIG. 2 301 to 324 See
FIG. 2 326 to 349 See FIG. 2 Leucine rich repeat carboxyl 359 to
405 See FIG. terminal domain (LRRCT) 2
[0106] Among the domains that occur in TANGO 325 protein are
leucine rich repeat (LRR) domains, including amino terminal and
carboxyl terminal LRR domains, and a P-loop domain. In one
embodiment, the protein of the invention has at least one domain
that is at least 55%, preferably at least about 65%, more
preferably at least about 75%, yet more preferably at least about
85%, and most preferably at least about 95% identical to one of
these domains. In another embodiment, the protein has at least one
amino terminal LRR domain, at least one carboxyl terminal LRR
domain, and a plurality of LRR domains interposed therebetween. In
yet another embodiment, the protein has at least one P-loop domain,
and a plurality (e.g., 2, 3, 4, or more) of the LRR domains
described herein in Table IV.
[0107] One or more LRR domains is present in a variety of proteins
involved in protein-protein interactions. Such proteins include,
for example, proteins involved in signal transduction, cell-to-cell
adhesion, cell-to-extracellular matrix adhesion, cell development,
DNA repair, RNA processing, and cellular molecular recognition
processes. Specialized LRR domains, designated LRR amino terminal
(LRRNT) domains and LRR carboxyl terminal (LRRCT) domains often
occur near the amino and carboxyl, respectively, ends of a series
of LRR domains. TANGO 325 protein has fourteen clustered LRR
domains, including (from the amino terminus toward the carboxyl
terminus of TANGO 325) an LRRNT domain, twelve LRR domains, and an
LRRCT domain. TANGO 325 is thus involved in one or more
physiological processes in which these other LRR domain-containing
proteins are involved, namely binding of cells with extracellular
proteins such as soluble extracellular proteins and cell surface
proteins of other cells.
[0108] The fact that TANGO 325 has an ATP/GTP-binding domain (i.e.,
a P-loop domain) within the extracellular domain of the protein
indicates that this protein is involved in transmembrane signaling
events. Considered in combination with the protein-binding LRR
domains present in the extracellular domain of TANGO 325 protein,
the presence of the ATP/GTP-binding domain indicates that TANGO 325
protein is capable of sensing extracellular proteins, including
ATP-binding proteins and GTP-binding proteins, and extracellular
nucleotides (e.g., ATP, ADP, and AMP). Thus, TANGO 325 protein is
involved in translating information (e.g., environmental conditions
or signaling molecules provided to the environment by other cells)
from the extracellular environment of the cell in which it is
expressed to one or more intracellular biochemical systems.
[0109] TANGO 325 exhibits amino acid sequence and nucleic acid
sequence homology with human Slit-1 protein. An alignment of the
amino acid sequences of TANGO 325 and human Slit-1 protein is shown
in FIGS. 2G to 2L. In this alignment (made using the ALIGN software
{Myers and Miller (1989) CABIOS, ver. 2.0}; pam120.mat scoring
matrix; gap opening penalty=12, gap extension penalty=4), the
proteins are 35.4% identical (i.e., 35.4% of the residues of TANGO
325 correspond to identical residues in Slit-1). An alignment of
the nucleotide sequences of the ORFs encoding TANGO 325 and human
Slit-1 protein is shown in FIGS. 2M-1 through 2M-18. The two ORFs
are 65.7% identical, as assessed using the same software and
parameters.
[0110] The signal peptide prediction program SIGNALP (Nielsen et
al. (1997) Protein Engineering 10:1-6) predicted that human TANGO
325 protein includes an approximately 31 (i.e., 29, 30, 31, 32, or
33) amino acid residue signal peptide (amino acid residues 1 to 31
of SEQ ID NO: 23; SEQ ID NO: 24) preceding the mature TANGO 325
protein (i.e., approximately amino acid residues 42 to 622 of SEQ
ID NO: 23; SEQ ID NO: 25). In one embodiment, human TANGO 325
protein includes an extracellular domain (amino acid residues 32 to
529 of SEQ ID NO: 23; SEQ ID NO: 26); a transmembrane domain (amino
acid residues 530 to 547 of SEQ ID NO: 23; SEQ ID NO: 27); and a
cytoplasmic domain (amino acid residues 548 to 622 of SEQ ID NO:
23; SEQ ID NO: 28). In an alternative embodiment, human TANGO 325
protein includes a cytoplasmic domain (amino acid residues 32 to
529 of SEQ ID NO: 23; SEQ ID NO: 26); a transmembrane domain (amino
acid residues 530 to 547 of SEQ ID NO: 23; SEQ ID NO: 27); and an
extracellular domain (amino acid residues 548 to 622 of SEQ ID NO:
23; SEQ ID NO: 28).
[0111] FIG. 2F depicts a hydrophobicity plot of human TANGO 325
protein. Relatively hydrophobic regions are above the dashed
horizontal line, and relatively hydrophilic regions are below the
dashed horizontal line. The hydrophobic region which corresponds to
amino acid residues 1 to 31 of SEQ ID NO: 23 is the signal sequence
of human TANGO 325 (SEQ ID NO: 24). The hydrophobic region which
corresponds to amino acid residues 530 to 547 of SEQ ID NO: 23 is
the transmembrane domain of human TANGO 325 (SEQ ID NO: 27). As
described elsewhere herein, relatively hydrophilic regions are
generally located at or near the surface of a protein, and are more
frequently effective immunogenic epitopes than are relatively
hydrophobic regions. For example, the region of human TANGO 325
protein from about amino acid residue 550 to about amino acid
residue 565 appears to be located at or near the surface of the
protein, while the region from about amino acid residue 168 to
about amino acid residue 185 appears not to be located at or near
the surface.
[0112] The predicted molecular weight of human TANGO 325 protein
without modification and prior to cleavage of the signal sequence
is about 70.3 kilodaltons. The predicted molecular weight of the
mature human TANGO 325 protein without modification and after
cleavage of the signal sequence is about 66.8 kilodaltons.
[0113] Northern analysis experiments indicated that mRNA
corresponding to the cDNA encoding TANGO 325 is expressed in the
tissues listed in Table V, wherein "+" indicates expression and "-"
indicates that expression could not be detected in the
corresponding tissue.
TABLE-US-00005 TABLE V Animal Tissue Relative Level of Expression
Human placenta + liver + kidney + pancreas + heart + brain -
skeletal muscle - lung -
[0114] Uses of TANGO 325 Nucleic Acids,
[0115] Polypeptides, and Modulators Thereof
[0116] TANGO 325 proteins are involved in disorders which affect
both tissues in which they are normally expressed and tissues in
which they are normally not expressed. Based on the observation
that TANGO 325 is expressed in human aortic endothelial tissue and
in placenta, liver, kidney, pancreas, and heart tissues, TANGO 325
protein is involved in one or more biological processes which occur
in these tissues. In particular, TANGO 325 is involved in
modulating growth, proliferation, survival, differentiation, and
activity of endothelial cells including, but not limited to,
vascular and cardiac (including valvular) endothelial cells of the
animal in which it is normally expressed. TANGO 325 also modulates
growth, proliferation, survival, differentiation, and activity of
placenta, liver, kidney, and pancreas cells. Thus, TANGO 325 has a
role in disorders which affect these cells and their growth,
proliferation, survival, differentiation, and activity. TANGO 325
polypeptides, nucleic acids, or modulators thereof can be used to
prognosticate, diagnose, inhibit, prevent, or alleviate one or more
of these disorders.
[0117] In one example, TANGO 325 polypeptides, nucleic acids, and
modulators thereof can be used to treat placental disorders, such
as the placental disorders described elsewhere in this disclosure.
TANGO 325 polypeptides, nucleic acids, or modulators thereof can be
used to prognosticate, diagnose, inhibit, prevent, or alleviate one
or more of these disorders.
[0118] In another example, TANGO 325 polypeptides, nucleic acids,
and modulators thereof, can be used to treat hepatic (i.e., liver)
disorders, such as the hepatic disorders described elsewhere in
this disclosure. TANGO 325 polypeptides, nucleic acids, or
modulators thereof can be used to prognosticate, diagnose, inhibit,
prevent, or alleviate one or more of these disorders.
[0119] In another example, TANGO 325 polypeptides, nucleic acids,
or modulators thereof, can be used to treat renal (i.e., kidney)
disorders, such as glomerular diseases (e.g., acute and chronic
glomerulonephritis, rapidly progressive glomerulonephritis,
nephrotic syndrome, focal proliferative glomerulonephritis,
glomerular lesions associated with systemic disease, such as
systemic lupus erythematosus, Goodpasture's syndrome, multiple
myeloma, diabetes, neoplasia, sickle cell disease, and chronic
inflammatory diseases), tubular diseases (e.g., acute tubular
necrosis and acute renal failure, polycystic renal disease,
medullary sponge kidney, medullary cystic disease, nephrogenic
diabetes, and renal tubular acidosis), tubulointerstitial diseases
(e.g., pyelonephritis, drug and toxin induced tubulointerstitial
nephritis, hypercalcemic nephropathy, and hypokalemic nephropathy)
acute and rapidly progressive renal failure, chronic renal failure,
nephrolithiasis, vascular diseases (e.g., hypertension and
nephrosclerosis, microangiopathic hemolytic anemia, atheroembolic
renal disease, diffuse cortical necrosis, and renal infarcts), and
tumors (e.g., renal cell carcinoma and nephroblastoma). TANGO 325
polypeptides, nucleic acids, or modulators thereof can be used to
prognosticate, diagnose, inhibit, prevent, or alleviate one or more
of these disorders.
[0120] Pancreatic disorders in which TANGO 325 can be involved
include the pancreatic disorders described elsewhere in this
disclosure. TANGO 325 polypeptides, nucleic acids, or modulators
thereof can be used to prognosticate, diagnose, inhibit, prevent,
or alleviate one or more of these disorders.
[0121] Because TANGO 325 exhibits expression in the heart, TANGO
325 nucleic acids, proteins, and modulators thereof can be used to
treat cardiovascular disorders. Examples of heart disorders with
which TANGO 325 can be involved include the cardiovascular
disorders described elsewhere in this disclosure. TANGO 325
polypeptides, nucleic acids, or modulators thereof can be used to
prognosticate, diagnose, inhibit, prevent, or alleviate one or more
of these disorders.
[0122] It is known that serum nucleotide levels (e.g., ATP) affect
cardiac contractility and vasomotor tone. Presence in TANGO 325 of
an ATP/GTP binding domain in the extracellular portion thereof
implicates this transmembrane protein in sensing of serum
nucleotide levels and transmission of the sensed level by
mechanisms not yet fully understood to myocytes underlying the
epithelium. Thus, TANGO 325 is involved in disorders such as
cardiovascular insufficiency, hypertension, hypotension, shock, and
the like.
[0123] Leukocytes are known to bind with vascular endothelial
surfaces in a reversible manner prior to penetrating the vascular
endothelium in route to an underlying tissue. Although a few
proteins have previously been implicated in the
leukocyte-endothelium binding process, the identities of all of the
proteins involved remain unknown. The presence of numerous LRR
domains on the exterior portion of TANGO 325 protein implicates
this protein in reversible binding of leukocytes to vascular
endothelium. Thus, TANGO 325 is involved in physiological processes
and disorders which involve leukocyte-endothelium binding. Such
processes and disorders include, by way of example, cellular
aspects of immune responses, autoimmune responses and disorders,
and migration of leukocytes to lymph nodes.
[0124] The aortic endothelium, as well as other vascular
endothelia, are known to be involved in detection of signals (e.g.,
metabolites, proteins, and the like) in the blood stream. Mammalian
Slit-1 protein is known to be involved in the human endocrine
system (Itoh et al. (1998) Brain Res. Mol. Brain. Res. 62:175-186).
Amino acid and nucleic acid sequence similarity of TANGO 325 with
human Slit-1 protein, as described herein, indicates that TANGO 325
is involved in sensing physiological signals by the endocrine
system. Thus, TANGO 325 is involved in one or more human endocrine
disorders such as pituitary disorders (e.g., diabetes insipidus),
thyroid disorders (e.g., hyperthyroidism, hypothyroidism, diabetes,
goiter, and growth and developmental disorders), adrenal disorders
(e.g., Addison's disease, Cushing's syndrome, hyperaldosteronism,
and pheochromocytoma), and the like.
[0125] Human Slit-1 protein is also known to be involved in
guidance of neuronal growth. The sequence similarity of TANGO 325
with Slit-1, as described herein, implicates TANGO 325 in growth,
development, maintenance, and regeneration of neurons. TANGO 325
can thus be used to prevent, diagnose, and treat a variety of
neurological disorders.
[0126] TANGO 364 cDNA clones (designated jthke076a05 and
jthkf069g11) encoding at least a portion of human TANGO 364 protein
were isolated from a human fetal skin cDNA library by computerized
sequence analysis of library ORFs which encode a signal sequence
(SPOT analysis). Human TANGO 364 protein is predicted by structural
analysis to be a transmembrane protein.
[0127] The full length of the cDNA encoding human TANGO 364 protein
(FIG. 3; SEQ ID NO: 31) is 3510 nucleotide residues. The ORF of
this cDNA, nucleotide residues 235 to 1764 of SEQ ID NO: 31 (i.e.,
SEQ ID NO: 32), encodes a 510-amino acid residue protein (FIG. 3;
SEQ ID NO: 33), corresponding to a 479-residue transmembrane
protein. TANGO 364 cDNA can exist in an alternatively-spliced form,
as listed in FIGS. 3G through 31. In this alternative form, TANGO
364 cDNA is 2510 nucleotide residues in length (SEQ ID NO: 41). The
ORF of this cDNA, nucleotide residues 2 to 898 of SEQ ID NO: 41
(i.e., SEQ ID NO: 42), encodes a 299-amino acid residue protein
(FIG. 3; SEQ ID NO: 43) which has the same sequence as the portions
of full length TANGO 364 protein indicated in the alignment (made
using the ALIGN software; pam120.mat scoring matrix; gap penalties
-12/-4) listed in FIGS. 3J and 3K. In the discussion which follows,
the full length and alternatively-spliced forms of TANGO 364
molecules are referred to individually and collectively as TANGO
364 molecules of the corresponding type (e.g., cDNA and
protein).
[0128] The invention thus includes purified human TANGO 364
protein, both in the form of the immature 510 amino acid residue
protein (SEQ ID NO: 33) and in the form of the mature 479 amino
acid residue protein (SEQ ID NO: 35). Mature human TANGO 364
proteins can be synthesized without the signal sequence polypeptide
at the amino terminus thereof, or it can be synthesized by
generating immature TANGO 364 protein and cleaving the signal
sequence therefrom.
[0129] The invention also includes nucleic acid molecules which
encode a polypeptide of the invention. Such nucleic acids include,
for example, a DNA molecule having the nucleotide sequence listed
in SEQ ID NO: 31 or some portion thereof, such as the portion which
encodes mature human TANGO 364 protein, immature human TANGO 364
protein, or a domain of human TANGO 364 protein. These nucleic
acids are collectively referred to as nucleic acids of the
invention.
[0130] TANGO 364 proteins and nucleic acid molecules encoding them
comprise a family of molecules having certain conserved structural
and functional features.
[0131] A common domain present in TANGO 364 proteins is a signal
sequence. In one embodiment, a TANGO 364 protein contains a signal
sequence corresponding to the portion of the protein from amino
acid residue 1 to about amino acid residue 31 of SEQ ID NO: 33 (SEQ
ID NO: 34). It is recognized that the carboxyl terminal boundary of
the signal sequence can be located one or two residues from the
residue identified above (i.e., at residue 29, 30, 31, 32, or 33 of
SEQ ID NO: 33). The signal sequence is cleaved during processing of
the mature protein.
[0132] TANGO 364 proteins can include an extracellular domain. The
human TANGO 364 protein extracellular domain is located from about
amino acid residue 32 to amino acid residue 345 of SEQ ID NO: 33
(i.e., the extracellular domain has the sequence SEQ ID NO:
36).
[0133] In addition, TANGO 364 can include a transmembrane domain.
In one embodiment, a TANGO 364 protein of the invention contains a
transmembrane domain corresponding to about amino acid residues 346
to 370 of SEQ ID NO:33 (i.e., the transmembrane domain has the
sequence SEQ ID NO: 37).
[0134] The present invention includes TANGO 364 proteins having a
cytoplasmic domain, particularly including proteins having a
carboxyl-terminal cytoplasmic domain. As used herein, a
"cytoplasmic domain" refers to a portion of a protein which is
localized to the cytoplasmic side of a lipid bilayer of a cell when
a nucleic acid encoding the protein is expressed in the cell. The
human TANGO 364 cytoplasmic domain is located from about amino acid
residue 371 to amino acid residue 510 of SEQ ID NO: 33 (i.e., the
cytoplasmic domain has the sequence SEQ ID NO: 38).
[0135] In an alternative embodiment, TANGO 364 proteins have a
cytoplasmic domain located from about amino acid residue 32 to
amino acid residue 345 of SEQ ID NO: 33 (i.e., the extracellular
domain has the sequence SEQ ID NO: 36); a transmembrane domain
corresponding to about amino acid residues 346 to 370 of SEQ ID NO:
33 (i.e., the transmembrane domain has the sequence SEQ ID NO: 37);
and an extracellular domain located from about amino acid residue
371 to amino acid residue 510 of SEQ ID NO: 33 (i.e., the
extracellular domain has the sequence SEQ ID NO: 38).
[0136] TANGO 364 proteins typically comprise a variety of potential
post-translational modification sites (often within an
extracellular domain), such as those described herein in Table VI,
as predicted by computerized sequence analysis of TANGO 364
proteins using amino acid sequence comparison software (comparing
the amino acid sequence of TANGO 364 with the information in the
PROSITE database {rel. 12.2; February, 1995} and the Hidden Markov
Models database {Rel. PFAM 3.3}).
TABLE-US-00006 TABLE VI Amino Acid Type of Potential Modification
Site Residues of Amino Acid or Domain SEQ ID NO: 33 Sequence
N-glycosylation site 281 to 284 NWTR 430 to 433 NSSC 489 to 492
NGTL Protein kinase C phosphorylation site 26 to 28 TGR 192 to 194
SSR 195 to 197 SFK 249 to 251 SVR 322 to 324 SSR 339 to 341 SGK 383
to 385 TQK 397 to 399 SIR 426 to 428 SLK 450 to 452 TVR 465 to 467
SGR 491 to 493 TLR Casein kinase II phosphorylation site 283 to 286
TRLD 322 to 325 SSRD 410 to 413 SQPE 426 to 429 SLKD 450 to 453
TVRE 456 to 459 TQTE N-myristoylation site 135 to 140 GSFQAR 162 to
167 GQGLTL 189 to 194 GTTSSR 218 to 223 GQPLTC 311 to 316 GIYVCH
354 to 359 GVIAAL 464 to 469 GSGRAE 477 to 482 GIKQAM 490 to 495
GTLRAK 500 to 505 GIYING Cell attachment sequence 55 to 57 RGD
Immunoglobulin-/major 45 to 129 See FIG. 3 histocompatibility
protein-like 162 to 225 (Ig-/MHC-like) domain 263 to 317
[0137] In various embodiments, the protein of the invention has at
least 1, 2, 4, 6, 10, 15, or 20 or more of the post-translational
modification sites described herein in Table VI.
[0138] Examples of additional domains present in human TANGO 364
protein include the RGD cell attachment sequence and Ig-/MHC-like
domains. In one embodiment, the protein of the invention has at
least one domain that is at least 55%, preferably at least about
65%, more preferably at least about 75%, yet more preferably at
least about 85%, and most preferably at least about 95% identical
to one of the Ig-/MHC-like domains described herein in Table VI.
Preferably, the protein of the invention has at least one
Ig-/MHC-like domain and one RGD cell attachment sequence.
[0139] Ig-/MHC-like domains are conserved among immunoglobulin (Ig)
constant (CL) regions and one of the three extracellular domains of
major histocompatibility proteins (MHC). Ig-/MHC-like domains are
involved in protein-to-protein and protein-to-ligand binding.
[0140] The signal peptide prediction program SIGNALP (Nielsen et
al. (1997) Protein Engineering 10:1-6) predicted that human TANGO
364 protein includes an approximately 31 amino acid signal peptide
(amino acid residues 1 to about 31 of SEQ ID NO: 33; SEQ ID NO: 34)
preceding the mature TANGO 364 protein (amino acid residues 32 to
510 of SEQ ID NO: 33; SEQ ID NO: 35). Human TANGO 364 protein
includes an extracellular domain (amino acid residues 32 to 345 of
SEQ ID NO: 33; SEQ ID NO: 36), a transmembrane domain (amino acid
residues 346 to 370 of SEQ ID NO: 33; SEQ ID NO: 37), and a
cytoplasmic domain (amino acid residues 371 to 510 of SEQ ID NO:
33; SEQ ID NO: 38).
[0141] FIG. 3F depicts a hydrophobicity plot of human TANGO 364
protein. Relatively hydrophobic regions are above the dashed
horizontal line, and relatively hydrophilic regions are below the
dashed horizontal line. The hydrophobic region which corresponds to
amino acid residues 1 to about 31 of SEQ ID NO: 33 is the signal
sequence of human TANGO 364 (SEQ ID NO: 34), and the hydrophobic
region which corresponds to amino acid residues 346 to 370 of SEQ
ID NO: 33 is the transmembrane region of TANGO 364 (SEQ ID NO: 37).
As described elsewhere herein, relatively hydrophilic regions are
generally located at or near the surface of a protein, and are more
frequently effective immunogenic epitopes than are relatively
hydrophobic regions. For example, the region of human TANGO 364
protein from about amino acid residue 371 to about amino acid
residue 410 appears to be located at or near the surface of the
protein, while the region from about amino acid residue 235 to
about amino acid residue 245 appears not to be located at or near
the surface.
[0142] The predicted molecular weight of human TANGO 364 protein
without modification and prior to cleavage of the signal sequence
is about 55.5 kilodaltons. The predicted molecular weight of the
mature human TANGO 364 protein without modification and after
cleavage of the signal sequence is about 52.1 kilodaltons.
[0143] TANGO 364 exhibits limited sequence similarity to numerous
cell surface proteins, including proteins which serve as cell
surface antigens, proteoglycans, and virus receptors.
[0144] Uses of TANGO 364 Nucleic Acids,
[0145] Polypeptides, and Modulators Thereof
[0146] TANGO 364 proteins are involved in disorders which affect
both tissues in which they are normally expressed and tissues in
which they are normally not expressed. Based on the observations
that cDNA corresponding to TANGO 364 occurs in a human fetal skin
cDNA library, it is evident that TANGO 364 protein is involved in
one or more biological processes which occur in skin tissues. In
particular, TANGO 364 is involved in modulating one or more of
growth, proliferation, survival, differentiation, activity,
morphology, and movement/migration of skin cells. Thus, TANGO 364
has a role in disorders which affect skin cells and one or more of
their growth, proliferation, survival, differentiation, activity,
morphology, and movement/migration, as well as the biological
function of skin.
[0147] There are several indications that TANGO 364 is a cell
surface protein which is involved in binding a ligand to the cell
which expresses the protein. For instance, presence in TANGO 364 of
an amino terminal extracellular domain that includes three
Ig-/MHC-like domains exemplifies the cell-surface ligand-binding
capability of TANGO 364. In addition, the amino acid sequence
similarity which TANGO 364 exhibits with respect to several other
cell surface ligand-binding proteins reinforces this view. Presence
in TANGO 364 of an Ig-/MHC-like domain indicates that the
corresponding region of TANGO 364 is structurally similar to this
conserved extracellular region, and that TANGO 364 is involved in
binding one or more of a ligand and a protein (including, for
example, a serum protein and a cell-surface protein of another
cell). Thus, molecules (e.g., antibodies and short peptides) which
are able to interact specifically with an Ig-/MHC-like domain of a
TANGO 364 protein can inhibit binding of TANGO 364 with its normal
ligand, thereby disrupting one or more physiological processes
associated with such binding. Furthermore, polypeptides (including,
for example, full-length TANGO 364 protein and polypeptides of at
least about 25 to 50 amino acid residues) which comprise all or
part of an Ig-/MHC-like domain of TANGO 364 can bind with one or
more of the normal ligands of TANGO 364, thereby replicating the
normal physiological effect of binding between TANGO 364 and the
ligand or inhibiting binding of endogenous TANGO 364 with the
ligand. Therefore, TANGO 364 protein, polypeptides having at least
one Ig-/MHC-like domain thereof, and molecules capable of
interacting with such a domain are useful for prognosticating,
diagnosing, treating, and inhibiting disorders associated with
aberrant binding of TANGO 364 and its normal ligand.
[0148] TANGO 364 is involved in binding an animal cell which
expresses it with one or more of a protein (e.g., an antibody, an
major histocompatibility protein, a lectin, or another cell surface
protein), a small molecule (e.g., a sugar, a hormone, or another
molecule having a molecular weight less than about 5000, 1000, or
500 or less Daltons), a component of the extracellular matrix
(e.g., a collagen protein), another cell of the same animal, a
bacterial or fungal cell, and a virus. Thus, TANGO 364 is involved
in modulating cell-to-cell adhesion, tissue and extracellular
matrix invasivity of cells, infectivity of cells by pathogens
(e.g., bacteria and viruses), endocrine signaling processes, tissue
developmental and organizational processes, and the like. Thus,
TANGO 364 is involved in disorders in which these physiological
processes are relevant.
[0149] Disorders associated with aberrant cell-to-cell adhesion
include tumor growth and metastasis, malformation or degradation of
neurological connections, autoimmune disorders, immune
insufficiency disorders, atherosclerosis, arteriosclerosis,
abnormal blood coagulation, and the like. Disorders associated with
tissue and extracellular matrix invasivity of cells include tumor
metastasis, osteoporosis, inflammation, and the like. Disorders
associated with pathogenic infections include infections associated
with bacteria, fungi, mycoplasmas, viruses, eukaryotic parasites,
and the like. Disorders associated with aberrant endocrine
signaling processes include, for example, diabetes mellitus,
hypoglycemia, glucagon disorders, pituitary disorders (e.g.,
diabetes insipidus), thyroid disorders (e.g., hyper- and
hypothyroidism), adrenal disorders (e.g., Addison's disease,
adrenal virilism, Cushing's syndrome, and hyperaldosteronism),
multiple endocrine neoplasias, and polyglandular deficiency
syndromes. Disorders associated with aberrant tissue developmental
and organizational processes include, for example, birth defects,
benign and malignant carcinogenesis, neurodegenerative disorders
(e.g., Alzheimer's disease), and the like. TANGO 364 proteins,
nucleic acids encoding them, and agents (e.g., antibodies,
peptides, and small molecules) that modulate activity or expression
of either of these can be used to prognosticate, diagnose, treat,
and inhibit one or more of these disorders.
TANGO 405
[0150] A cDNA clone (designated jthLa152h06) encoding at least a
portion of human TANGO 405 protein was isolated from a human mixed
lymphocyte reaction cDNA library. A corresponding murine cDNA
(designated jtmMa025a11) was isolated from a long-term bone marrow
cDNA library. Human and murine TANGO 405 proteins are secreted
proteins.
[0151] The full length of the cDNA encoding human TANGO 405 protein
(FIG. 4; SEQ ID NO: 51) is 3114 nucleotide residues in length. The
open reading frame (ORF) of this cDNA, nucleotide residues 154 to
780 of SEQ ID NO: 51 (i.e., SEQ ID NO: 52), encodes a 209-amino
acid residue protein (FIG. 4; SEQ ID NO: 53), corresponding to a
161-residue secreted protein.
[0152] The invention thus includes purified human TANGO 405
protein, both in the form of the immature 209 amino acid residue
protein (SEQ ID NO: 53) and in the form of the mature 161 amino
acid residue protein (SEQ ID NO: 55). The invention also includes
purified murine TANGO 405 protein, both in the form of the immature
178-amino acid residue protein (SEQ ID NO: 63) and in the form of
the mature, secreted 136-amino acid residue protein (SEQ ID NO:
65). Mature human or murine TANGO 405 protein can be synthesized
without the signal sequence polypeptide at the amino terminus
thereof, or they can be synthesized by generating immature TANGO
405 protein and cleaving the signal sequence therefrom.
[0153] The invention also includes nucleic acid molecules which
encode a polypeptide of the invention. Such nucleic acids include,
for example, a DNA molecule having the nucleotide-sequence listed
in SEQ ID NO: 51 or some portion thereof or SEQ ID NO: 61 or some
portion thereof, such as the portion which encodes mature human or
murine TANGO 405 protein, immature human or murine TANGO 405
protein, or a domain of human or murine TANGO 405 protein. These
nucleic acids are collectively referred to as nucleic acids of the
invention.
[0154] TANGO 405 proteins and nucleic acid molecules encoding them
comprise a family of molecules having certain conserved structural
and functional features.
[0155] A common domain present in TANGO 405 proteins is a signal
sequence. In one embodiment, a TANGO 405 protein contains a signal
sequence corresponding to the portion of the protein from amino
acid residue 1 to about amino acid residue 48 of SEQ ID NO: 53 (SEQ
ID NO: 54) or to the portion of the protein from amino acid residue
1 to about amino acid residue 42 of SEQ ID NO: 63 (SEQ ID NO: 64).
It is recognized that the carboxyl terminal boundary of the signal
sequence can be located one or two residues from the residue
identified above (i.e., at residue 46, 47, 48, 49, or 50 of SEQ ID
NO: 53 or at residue 40, 41, 42, 43 or 44 of SEQ ID NO: 63). The
signal sequence is cleaved during processing of the mature
protein.
[0156] TANGO 405 proteins typically comprise a variety of potential
post-translational modification sites (often within an
extracellular domain), such as those described herein in Table VII
(for human TANGO 405) and VIII (for murine TANGO 405), as predicted
by computerized sequence analysis of TANGO 405 proteins using amino
acid sequence comparison software (comparing the amino acid
sequence of TANGO 405 with the information in the PROSITE database
{rel. 12.2; February, 1995} and the Hidden Markov Models database
{Rel. PFAM 3.3}).
TABLE-US-00007 TABLE VII Amino Acid Amino Type of Potential
Residues of Acid Modification Site or Domain SEQ ID NO: 53 Sequence
N-glycosylation site 131 to 134 NESF 170 to 173 NHSA cAMP- or
cGMP-dependent 52 to 55 KRLS protein kinase phosphory- 197 to 200
RRNS lation site Protein kinase C phosphory- 10 to 12 TEK lation
site 17 to 19 SLR 50 to 52 TGK 82 to 84 SWK 196 to 198 TRR Casein
kinase II phosphory- 46 to 49 TYGE lation site 94 to 97 SSEE 101 to
104 SKSE 119 to 122 TEAE 155 to 158 TPYE 200 to 203 SICE Tyrosine
kinase phosphory- 52 to 60 KRLSELHSY lation site N-myristoylation
site 25 to 30 GISIAL 77 to 82 GCCPAS Amidation site 50 to 53 TGKR
C-type lectin domain 176 to 202 See FIG. signature 4 C-type lectin
domain 105 to 202 See FIG. 4
TABLE-US-00008 TABLE VIII Amino Acid Amino Type of Potential
Residues of Acid Modification Site or Domain SEQ ID NO: 63 Sequence
N-glycosylation site 136 to 139 NESL 155 to 158 NGSM Protein kinase
C phosphory- 20 to 22 TLR lation site 54 to 56 SRR 77 to 79 SEK 99
to 101 STK 162 to 164 SVK Casein kinase II phosphory- 99 to 102
STKE lation site 106 to 109 STSE 124 to 127 TEAE Tyrosine kinase
phosphory- 55 to 63 RRLYELHTY lation site N-myristoylation site 16
to 21 GVCWTL 73 to 78 GTMVSE 82 to 87 GCCPNH C-type lectin domain
110 to 180 See FIG. 4
[0157] In various embodiments, the protein of the invention has at
least 1, 2, 4, 6, 10, 15, or 20 or more of the post-translational
modification sites described herein in Tables VII and VIII.
[0158] Examples of additional domains present in human and murine
TANGO 405 protein include a C-type lectin domain and a
corresponding signature sequence. In one embodiment, the protein of
the invention has a C-type lectin domain or signature sequence that
is at least 55%, preferably at least about 65%, more preferably at
least about 75%, yet more preferably at least about 85%, and most
preferably at least about 95% identical to one of those described
herein in Tables VII and VIII.
[0159] C-type lectin domains are conserved among proteins (e.g.,
animal lectins) which are involved in calcium-dependent binding of
carbohydrates, although it has recently been recognized that these
domains can also be involved in binding of proteins (Drickamer,
1988, J. Biol. Chem. 263:9557-9560; Drickamer, 1993, Prog. Nucl.
Acid Res. Mol. Biol. 45:207-232; Drickamer, 1993, Curr. Opin.
Struct. Biol. 3:393-400). C-type lectins and their relevant
properties are described in greater in P.C.T. Publication No. WO
98/28332, which, as with all references cited herein, is
incorporated by reference.
[0160] In PCT Publication No. WO 98/28332, a cDNA encoding murine
protein, designated dectin-2, was isolated from dendritic cells and
described. Human and murine TANGO 405 proteins exhibit amino acid
sequence homology with murine dectin-2. As indicated in the
alignment in FIG. 4M (made using the ALIGN software; pam120.mat
scoring matrix; gap penalties -12/-4), human TANGO 405 exhibits
about 89.0% sequence identity with murine dectin-2. As indicated in
the alignment in FIG. 4L (made using the ALIGN software; pam120.mat
scoring matrix; gap penalties -12/-4), murine TANGO 405 exhibits
about 70.3% sequence identity with murine dectin-2.
[0161] Another embodiment of a murine TANGO 405 cDNA is shown in
FIGS. 4N to 4P (the cDNA having the sequence SEQ ID NO: 71 and the
ORF having the nucleotide sequence SEQ ID NO: 72). In this
embodiment murine TANGO 405 includes a translational frame shift,
and the amino acid sequence (SEQ ID NO: 73) of murine TANGO 405 is
identical to the amino acid sequence reported for murine dectin-2.
These data further confirm that human TANGO 405 is the human
ortholog of murine dectin-2.
[0162] The signal peptide prediction program SIGNALP (Nielsen et
al. (1997) Protein Engineering 10:1-6) predicted that human TANGO
405 protein includes an approximately 48 amino acid signal peptide
(amino acid residues 1 to about 48 of SEQ ID NO: 53; SEQ ID NO: 54)
preceding the mature TANGO 405 protein (amino acid residues 49 to
209 of SEQ ID NO: 53; SEQ ID NO: 55). It is recognized that both
human and murine TANGO 405 can, at least transiently, exist in an
integral membrane form, at least until cleavage of the
corresponding signal sequence (i.e., either during or following
translation of the complete TANGO 405 protein).
[0163] FIG. 4D depicts a hydrophobicity plot of human TANGO 405
protein. Relatively hydrophobic regions are above the dashed
horizontal line, and relatively hydrophilic regions are below the
dashed horizontal line. The hydrophobic region which corresponds to
amino acid residues 1 to about 48 of SEQ ID NO: 53 is the signal
sequence of human TANGO 405 (SEQ ID NO: 54). As described elsewhere
herein, relatively hydrophilic regions are generally located at or
near the surface of a protein, and are more frequently effective
immunogenic epitopes than are relatively hydrophobic regions. For
example, the region of human TANGO 405 protein from about amino
acid residue 90 to about amino acid residue 105 appears to be
located at or near the surface of the protein, while the region
from about amino acid residue 110 to about amino acid residue 120
appears not to be located at or near the surface.
[0164] The predicted molecular weight of human TANGO 405 protein
without modification and prior to cleavage of the signal sequence
is about 24.0 kilodaltons. The predicted molecular weight of the
mature human TANGO 405 protein without modification and after
cleavage of the signal sequence is about 18.6 kilodaltons.
[0165] The full length of the cDNA encoding murine TANGO 405
protein (FIG. 4; SEQ ID NO: 61) is 821 nucleotide residues,
although this cDNA sequence is incomplete. The ORF of this cDNA,
nucleotide residues 174 to 707 of SEQ ID NO: 61 (i.e., SEQ ID NO:
62), encodes a protein comprising at least 178 amino acid residues
(FIG. 4; SEQ ID NO: 63).
[0166] The signal peptide prediction program SIGNALP (Nielsen et
al. (1997) Protein Engineering 10:1-6) predicted that murine TANGO
405 protein includes an approximately 42 amino acid signal peptide
(amino acid residues 1 to about 42 of SEQ ID NO: 63; SEQ ID NO: 64)
preceding the mature TANGO 405 protein (amino acid residues 43 to
178 of SEQ ID NO: 63; SEQ ID NO: 65). Murine TANGO 405 protein is a
secreted protein.
[0167] FIG. 4G depicts a hydrophobicity plot of murine TANGO 405
protein. Relatively hydrophobic regions are above the dashed
horizontal line, and relatively hydrophilic regions are below the
dashed horizontal line. The hydrophobic region which corresponds to
amino acid residues 1 to about 42 of SEQ ID NO: 63 is the signal
sequence of murine TANGO 405 (SEQ ID NO: 64). As described
elsewhere herein, relatively hydrophilic regions are generally
located at or near the surface of a protein, and are more
frequently effective immunogenic epitopes than are relatively
hydrophobic regions. For example, the region of murine TANGO 405
protein from about amino acid residue 95 to about amino acid
residue 110 appears to be located at or near the surface of the
protein, while the region from about amino acid residue 110 to
about amino acid residue 120 appears not to be located at or near
the surface
[0168] The predicted molecular weight of murine TANGO 405 protein
without modification and prior to cleavage of the signal sequence
is about 20.0 kilodaltons. The predicted molecular weight of the
mature murine TANGO 405 protein without modification and after
cleavage of the signal sequence is about 25.3 kilodaltons.
[0169] Human and murine TANGO 405 proteins exhibit considerable
sequence similarity, as indicated herein in FIG. 4H. FIG. 4H
depicts an alignment of human and murine TANGO 405 amino acid
sequences (SEQ ID NOs: 53 and 63, respectively). In this alignment
(made using the ALIGN software {Myers and Miller (1989) CABIOS,
ver. 2.0}; pam120.mat scoring matrix; gap penalties -12/-4), the
proteins are 51.7% identical in the overlapping region (i.e., amino
acid residues 1-209 of SEQ ID NO: 53 and amino acid residues 1-178
of SEQ ID NO: 63). The human and murine ORFs encoding TANGO 405 are
74.5% identical in the 541 nucleotide residue overlapping region,
as assessed using the same software and parameters and as indicated
in FIGS. 4I through 4K. The nucleotide sequences encoding human and
murine TANGO 405 (i.e., SEQ ID NOs: 51 and 61) are about 71.2%
identical in the 838 nucleotide residue overlapping region, as
assessed using the LALIGN software (Myers and Miller (1989) CABIOS,
ver. 2.0; pam120.mat scoring matrix; gap penalties -12/-4).
[0170] Uses of TANGO 405 Nucleic Acids,
[0171] Polypeptides, and Modulators Thereof
[0172] TANGO 405 proteins are involved in disorders which affect
both tissues in which they are normally expressed and tissues in
which they are normally not expressed. Based on the observations
that cDNA corresponding to TANGO 405 occurs in a human mixed
lymphocyte reaction cDNA library and in a murine long-term bone
marrow cDNA library, it is evident that TANGO 405 protein is
involved in one or more biological processes which occur in these
tissues (i.e., in blood-related tissues, such as tissues containing
lymphocytes). In particular, TANGO 405 is involved in modulating
one or more of growth, proliferation, survival, differentiation,
activity, morphology, and movement/migration of cells of these
tissues. TANGO 405 is involved in modulating the structure of
extracellular matrix which contacts or is in fluid communication
with cells of these tissues. Thus, TANGO 405 has a role in
disorders which affect these cells and one or more of their growth,
proliferation, survival, differentiation, activity, morphology, and
movement/migration, as well as the biological function of tissues
comprising one or more of these types of cells.
[0173] Presence of a C-type lectin domain in TANGO 405 is an
indication that this protein is capable of specifically recognizing
particular surfaces, such as the surface of cells of a particular
type. Further supportive of this observation is the fact that human
TANGO 405 protein exhibits significant sequence homology with
murine dectin-2 protein. Murine dectin-2 has been shown to be
expressed by murine dendritic cells, and has also been shown to be
involved in activation of naive T cells. Murine dectin-2 can also
be involved in inflammatory and non-T cell-mediated immune
responses. Thus, human and murine TANGO 405 are also involved in
activating or inhibiting one or more types of lymphocytes, thereby
modulating T cell-mediated immune responses, non-T cell-mediated
immune responses, inflammatory responses, and other components of
the immune response in mammals. It is recognized that the amino
acid sequence differences among murine dectin-2, human TANGO 405,
and murine TANGO 405 can lead to different lymphocyte-activating
capacities for these three proteins. Human and murine TANGO 405
proteins are involved both in normal activation of lymphocytes
(e.g., in response to the presence of a pathogen in a tissue) and
in aberrant activation of lymphocytes (e.g., as in auto-immune and
immune inflammatory disorders (e.g., asthma), in disorders
characterized by an insufficient immune response, and in disorders
characterized by non-controlled proliferation of lymphocytes).
TANGO 405 proteins are thus involved in a variety of disorders
relating to aberrant lymphocyte activation or proliferation.
Examples of disorders include leukemias (e.g., ALL, CML, CLL, and
myelodysplastic syndrome), lymphomas (e.g., Hodgkin's disease,
non-Hodgkin's lymphoma, Burkitt's lymphoma, and mycosis fungoides),
plasma cell dyscrasias, auto-immune disorders such as multiple
sclerosis, bacterial and viral infections (e.g., acquired immune
deficiency syndrome), leukopenias, eosinophilic disorders such as
idiopathic hypereosinophilic syndrome, and the like. TANGO 405
proteins, nucleic acids encoding them, and agents that modulate
activity or expression of either of these can be used to
prognosticate, diagnose, treat, and inhibit one or more of these
disorders.
[0174] M019 (Also Designated TANGO 533)
[0175] A cDNA encoding at least a portion of human M019 protein was
isolated from a human adipose tissue cDNA library. The human M019
protein is predicted by structural analysis to be a secreted
protein.
[0176] The full length of the cDNA encoding human M019 protein
(FIG. 5; SEQ ID NO: 81) is 1202 nucleotide residues. The ORF of
this cDNA, nucleotide residues 331 to 585 of SEQ ID NO: 81 (FIG. 5;
SEQ ID NO: 82), encodes a 85-amino acid secreted protein (FIG. 5;
SEQ ID NO: 83).
[0177] The invention thus includes purified human M019 protein,
both in the form of the immature 85 amino acid residue protein (SEQ
ID NO: 83) and in the form of the mature 62 amino acid residue
protein (SEQ ID NO: 85). Mature human M019 protein can be
synthesized without the signal sequence polypeptide at the amino
terminus thereof, or it can be synthesized by generating immature
M019 protein and cleaving the signal sequence therefrom.
[0178] The invention also includes nucleic acid molecules which
encode a polypeptide of the invention. Such nucleic acids include,
for example, a DNA molecule having the nucleotide sequence listed
in SEQ ID NO: 81 or some portion thereof, such as the portion which
encodes mature M019 protein, immature M019 protein, or a domain of
M019 protein. These nucleic acids are collectively referred to as
nucleic acids of the invention.
[0179] M019 proteins and nucleic acid molecules encoding them
comprise a family of molecules having certain conserved structural
and functional features. Each of these molecules is included in the
invention. As used herein, the term "family" is intended to mean
two or more proteins or nucleic acid molecules having a common or
similar domain structure and having sufficient amino acid or
nucleotide sequence identity as defined herein. Family members can
be from either the same or different species. For example, a family
can comprise two or more proteins of human origin, or can comprise
one or more proteins of human origin and one or more of non-human
origin.
[0180] A common domain present in M019 proteins is a signal
sequence. In one embodiment, a M019 protein contains a signal
sequence corresponding to amino acid residues 1 to 23 of SEQ ID NO:
83 (SEQ ID NO: 84). The signal sequence is cleaved during
processing of the mature protein.
[0181] M019 proteins are secreted proteins, and thus include an
`extracellular domain,` both in the mature protein (i.e., wherein
the entire mature protein is an `extracellular domain`) and in the
immature protein (e.g., wherein the signal sequence, residues 1-23
of SEQ ID NO: 83, is embedded in the membrane prior to cleavage,
and the remainder of the protein, about residues 24-85 is
extracellular). As used herein, an "extracellular domain" refers to
a portion of a protein which is localized to the non-cytoplasmic
side of a lipid bilayer of a cell when a nucleic acid encoding the
protein is expressed in the cell.
[0182] M019 proteins typically comprise a variety of potential
post-translational modification sites (often within an
extracellular domain), such as those described herein in Table IX,
as predicted by computerized sequence analysis of M019 proteins
using amino acid sequence comparison software (comparing the amino
acid sequence of M019 with the information in the PROSITE database
{rel. 12.2; February, 1995} and the Hidden Markov Models database
{Rel. PFAM 3.3}). In certain embodiments, a protein of the
invention has at least 1, 2, 3, or all 4 of the post-translational
modification sites listed in Table IX.
TABLE-US-00009 TABLE IX Amino Acid Amino Type of Potential Residues
of Acid Modification Site or Domain SEQ ID NO: 83 Sequence Protein
kinase C phosphory- 47 to 49 SNR lation site 75 to 77 TMK Casein
kinase II phosphory- 47 to 50 SNRE lation site N-myristoylation
site 34 to 49 GQDSNL
[0183] M019 exhibits no significant amino acid sequence similarity
with any known protein. Thus, M019 appears to be a novel
protein.
[0184] FIG. 5C depicts a hydrophobicity plot of human M019 protein.
Relatively hydrophobic regions are above the dashed horizontal
line, and relatively hydrophilic regions are below the dashed
horizontal line. The hydrophobic region which corresponds to amino
acid residues 1 to 23 of SEQ ID NO: 83 is the signal sequence of
human M019 (SEQ ID NO: 84). As described elsewhere herein,
relatively hydrophilic regions are generally located at or near the
surface of a protein, and are more frequently effective immunogenic
epitopes than are relatively hydrophobic regions. For example, the
region of human M019 protein from about amino acid residue 63 to
about amino acid residue 80 appears to be located at or near the
surface of the protein, while the region from about amino acid
residue 55 to about amino acid residue 60 appears not to be located
at or near the surface.
[0185] Uses of M019 Nucleic Acids,
[0186] Polypeptides, and Modulators Thereof
[0187] M019 proteins are involved in disorders which affect both
tissues in which they are normally expressed and tissues in which
they are normally not expressed. Based on the observation that M019
is expressed in adipose tissue, M019 protein is involved in one or
more biological processes which occur in these tissues and in
disorders which affect adipose tissue. Such disorder include, for
example, obesity, hypercholesterolemia, hyperlipidemia,
hyperlipoproteinemia, diabetes, stroke, liver fibrosis,
atherosclerosis, arteriosclerosis, and coronary artery disease.
[0188] By virtue of its size and the presence of a pair of cysteine
residues in its mature form, M019 resembles chemokine molecules,
and it therefore believed to be involved in modulating adipose
tissue processes involving interaction of cells (e.g., leukocytes)
and proteins (e.g., lipoproteins) with the surface of adipose
tissue cells. Such processes include, for example, uptake, release,
metabolism, and storage of lipids (e.g., triglycerides),
cholesterol, lipoproteins, and the like.
[0189] M019 exhibits limited sequence similarity with pancreatic
proteins, indicating that this protein is involved in physiological
processes of the pancreas and in pancreatic disorders, as well as
other disorders. Pancreatic disorders in which M019 can be involved
include those pancreatic disorders described elsewhere in this
disclosure.
[0190] Tables A and B summarize sequence data corresponding to the
nucleic acids and proteins disclosed herein.
TABLE-US-00010 TABLE A SEQ ID NOs Pro- Depicted in ATCC .RTM.
Protein Designation cDNA ORF tein Figure # Accession # human TANGO
273 1 2 3 1 207185 murine TANGO 273 11 12 13 1 207221 human TANGO
325 21 22 23 2 PTA-147 human TANGO 364 31 32 33 3 PTA-425 human
TANGO 364 41 42 43 3 PTA-425 (alternative form) human TANGO 405 51
52 53 4 PTA-424 murine TANGO 405 61 62 63 4 murine TANGO 405 71 72
73 4 (alternative form) human M019 81 82 83 5
TABLE-US-00011 TABLE B Signal Extracellular Transmembrane
Cytoplasmic Sequence.sup.1 Mature Protein Domain(s).sup.2 Domain(s)
Domain(s).sup.2 Protein Desig. SEQ ID NOs hum. TANGO 273 1 to 22 4
23 to 172 5 23 to 60 6 61 to 81 7 82 to 172 8 mur. TANGO 273 1 to
22 14 23 to 172 15 23 to 60 16 61 to 81 17 82 to 172 18 hum. TANGO
325 1 to 31 24 32 to 622 25 32 to 529 26 530 to 547 27 548 to 622
28 hum. TANGO 364 1 to 31 34 32 to 510 35 32 to 345 36 346 to 370
37 371 to 510 38 hum. TANGO 405 1 to 48 54 49 to 209 55 49 to 209
55 N/A N/A mur. TANGO 405 1 to 52 64 53 to 178 65 53 to 178 65 N/A
N/A hum. M019 1 to 23 84 24 to 82 85 24 to 82 85 Amino Acid
Residues Notes for Table B: .sup.1It is recognized that the
carboxyl terminal boundary of the signal sequence can be .+-.1 or 2
residues from that indicated. .sup.2It is recognized that
`extracellular` and cytoplasmic` domains can have the opposite
orientation in certain embodiments, as described herein.
[0191] Various aspects of the invention are described in further
detail in the following subsections.
I. Isolated Nucleic Acid Molecules
[0192] One aspect of the invention pertains to isolated nucleic
acid molecules that encode a polypeptide of the invention or a
biologically active portion thereof, as well as nucleic acid
molecules sufficient for use as hybridization probes to identify
nucleic acid molecules encoding a polypeptide of the invention and
fragments of such nucleic acid molecules suitable for use as PCR
primers for the amplification or mutation of nucleic acid
molecules. As used herein, the term "nucleic acid molecule" is
intended to include DNA molecules (e.g., cDNA or genomic DNA) and
RNA molecules (e.g., mRNA) and analogs of the DNA or RNA generated
using nucleotide analogs. The nucleic acid molecule can be
single-stranded or double-stranded.
[0193] An "isolated" nucleic acid molecule is one which is
separated from other nucleic acid molecules which are present in
the natural source of the nucleic acid molecule. Preferably, an
"isolated" nucleic acid molecule is free of sequences (preferably
protein-encoding sequences) which naturally flank the nucleic acid
(i.e., sequences located at the 5' and 3' ends of the nucleic acid)
in the genomic DNA of the organism from which the nucleic acid is
derived. For example, in various embodiments, the isolated nucleic
acid molecule can contain less than about 5, 4, 3, 2, 1, 0.5, or
0.1 kilobases of nucleotide sequences which naturally flank the
nucleic acid molecule in genomic DNA of the cell from which the
nucleic acid is derived. Moreover, an "isolated" nucleic acid
molecule, such as a cDNA molecule, can be substantially free of
other cellular material, or culture medium when produced by
recombinant techniques, or substantially free of chemical
precursors or other chemicals when chemically synthesized.
[0194] A nucleic acid molecule of the present invention, e.g., a
nucleic acid molecule having the nucleotide sequence of all or a
portion of any of SEQ ID NOs: 1, 2, 11, 12, 21, 22, 31, 32, 41, 42,
51, 52, 61, 62, 71, 72, 81, 82, and the nucleotide sequence of any
of the clones deposited as ATCC.RTM. Accession numbers 207185,
207221, PTA-147, PTA-425, and PTA-424, or a complement thereof, or
which has a nucleotide sequence comprising one of these sequences,
can be isolated using standard molecular biology techniques and the
sequence information provided herein. Using all or a portion of the
nucleic acid sequences of any of SEQ ID NOs: 1, 2, 11, 12, 21, 22,
31, 32, 41, 42, 51, 52, 61, 62, 71, 72, 81, 82, and the nucleotide
sequence of any of the clones deposited as ATCC.RTM. Accession
numbers 207185, 207221, PTA-147, PTA-425, and PTA-424 as a
hybridization probe, nucleic acid molecules of the invention can be
isolated using standard hybridization and cloning techniques (e.g.,
as described in Sambrook et al., Eds., Molecular Cloning: A
Laboratory Manual, 2nd ed., Cold Spring Harbor Laboratory, Cold
Spring Harbor Laboratory Press, Cold Spring Harbor, N.Y.,
1989).
[0195] A nucleic acid molecule of the invention can be amplified
using cDNA, mRNA, or genomic DNA as a template and appropriate
oligonucleotide primers according to standard PCR amplification
techniques. The nucleic acid so amplified can be cloned into an
appropriate vector and characterized by DNA sequence analysis.
Furthermore, oligonucleotides corresponding to all or a portion of
a nucleic acid molecule of the invention can be prepared by
standard synthetic techniques, e.g., using an automated DNA
synthesizer.
[0196] In another embodiment, an isolated nucleic acid molecule of
the invention comprises a nucleic acid molecule which is a
complement of the nucleotide sequence of any of SEQ ID NOs: 1, 2,
11, 12, 21, 22, 31, 32, 41, 42, 51, 52, 61, 62, 71, 72, 81, 82, and
the nucleotide sequence of any of the clones deposited as ATCC.RTM.
Accession numbers 207185, 207221, PTA-147, PTA-425, and PTA-424, or
a portion thereof. A nucleic acid molecule which is complementary
to a given nucleotide sequence is one which is sufficiently
complementary to the given nucleotide sequence that it can
hybridize with the given nucleotide sequence thereby forming a
stable duplex.
[0197] Moreover, a nucleic acid molecule of the invention can
comprise a portion of a nucleic acid sequence encoding a full
length polypeptide of the invention, such as a fragment which can
be used as a probe or primer or a fragment encoding a biologically
active portion of a polypeptide of the invention. The nucleotide
sequence determined from cloning one gene allows generation of
probes and primers designed for identifying and/or cloning homologs
in other cell types, e.g., from other tissues, as well as homologs
from other mammals. The probe/primer typically comprises
substantially purified oligonucleotide. The oligonucleotide
typically comprises a region of nucleotide sequence that hybridizes
under stringent conditions with at least about 15, preferably about
25, more preferably about 40, 60, 80, 100, 150, 200, 250, 300, 350,
400, 450, 550, 650, 700, 800, 900, 1000, 1200, 1400, 1410, 1600,
1800, 2000, 2200, 2400, 2600, 2800, 3000, or 3500 or more
consecutive nucleotides of the sense or anti-sense sequence of any
of SEQ ID NOs: 1, 2, 11, 12, 21, 22, 31, 32, 41, 42, 51, 52, 61,
62, 71, 72, 81, 82, and the nucleotide sequence of any of the
clones deposited as ATCC.RTM. Accession numbers 207185, 207221,
PTA-147, PTA-425, and PTA-424, or of a naturally occurring mutant
of any of these sequences.
[0198] Probes based on the sequence of a nucleic acid molecule of
the invention can be used to detect transcripts or genomic
sequences encoding the same protein molecule encoded by a selected
nucleic acid molecule. The probe comprises a label group attached
thereto, e.g., a radioisotope, a fluorescent compound, an enzyme,
or an enzyme co-factor. Such probes can be used as part of a
diagnostic test kit for identifying cells or tissues which
aberrantly express the protein, such as by measuring levels of a
nucleic acid molecule encoding the protein in a sample of cells
from a subject, e.g., detecting mRNA levels or determining whether
a gene encoding the protein has been mutated or deleted.
[0199] A nucleic acid fragment encoding a biologically active
portion of a polypeptide of the invention can be prepared by
isolating a portion of one of SEQ ID NOs: 2, 12, 22, 32, 42, 52,
62, 72, and 82 expressing the encoded portion of the polypeptide
protein (e.g., by recombinant expression in vitro), and assessing
the activity of the encoded portion of the polypeptide.
[0200] The invention further encompasses nucleic acid molecules
that differ from the nucleotide sequence of any of SEQ ID NOs: 1,
2, 11, 12, 21, 22, 31, 32, 41, 42, 51, 52, 61, 62, 71, 72, 81, 82,
and the nucleotide sequence of any of the clones deposited as
ATCC.RTM. Accession numbers 207185, 207221, PTA-147, PTA-425, and
PTA-424, due to degeneracy of the genetic code and thus encode the
same protein as that encoded by the nucleotide sequence of one of
SEQ ID NOs: 2, 12, 22, 32, 42, 52, 62, 72, and 82.
[0201] In addition to the nucleotide sequences of one of SEQ ID
NOs: 2, 12, 22, 32, 42, 52, 62, 72, and 82, it will be appreciated
by those skilled in the art that DNA sequence polymorphisms that
lead to changes in the amino acid sequence can exist within a
population (e.g., the human population). Such genetic polymorphisms
can exist among individuals within a population due to natural
allelic variation. An allele is one of a group of genes which occur
alternatively at a given genetic locus.
[0202] To determine the percent identity of two amino acid
sequences or of two nucleic acids, the sequences are aligned for
optimal comparison purposes (e.g., gaps can be introduced in the
sequence of a first amino acid or nucleic acid sequence for optimal
alignment with a second amino or nucleic acid sequence). The amino
acid residues or nucleotides at corresponding amino acid positions
or nucleotide positions are then compared. When a position in the
first sequence is occupied by the same amino acid residue or
nucleotide as the corresponding position in the second sequence,
then the molecules are identical at that position. The percent
identity between the two sequences is a function of the number of
identical positions shared by the sequences (i.e., % identity=# of
identical positions/total # of positions {e.g., overlapping
positions}.times.100). In one embodiment, the two sequences are the
same length.
[0203] The determination of percent identity between two sequences
can be accomplished using a mathematical algorithm. A preferred,
non-limiting example of a mathematical algorithm utilized for the
comparison of two sequences is the algorithm of Karlin and Altschul
(1990) Proc. Natl. Acad. Sci. USA 87:2264-2268, modified as in
Karlin and Altschul (1993) Proc. Natl. Acad. Sci. USA 90:5873-5877.
Such an algorithm is incorporated into the NBLAST and XBLAST
programs of Altschul, et al. (1990) J. Mol. Biol. 215:403-410.
BLAST nucleotide searches can be performed with the NBLAST program,
score=100, wordlength=12 to obtain nucleotide sequences homologous
to a nucleic acid molecules of the invention. BLAST protein
searches can be performed with the XBLAST program, score=50,
wordlength=3 to obtain amino acid sequences homologous to a protein
molecules of the invention. To obtain gapped alignments for
comparison purposes, Gapped BLAST can be utilized as described in
Altschul et al. (1997) Nucleic Acids Res. 25:3389-3402.
Alternatively, PSI-Blast can be used to perform an iterated search
which detects distant relationships between molecules (Id.). When
utilizing BLAST, Gapped BLAST, and PSI-Blast programs, the default
parameters of the respective programs (e.g., XBLAST and NBLAST) can
be used.
[0204] Another preferred, non-limiting example of a mathematical
algorithm utilized for the comparison of sequences is the local
homology algorithm of Smith and Waterman (Advances in Applied
Mathematics 2: 482-489 (1981)). Such an algorithm is incorporated
into the BestFit program, which is part of the Wisconsin.TM.
package, and is used to find the best segment of similarity between
two sequences. BestFit reads a scoring matrix that contains values
for every possible GCG symbol match. The program uses these values
to construct a path matrix that represents the entire surface of
comparison with a score at every position for the best possible
alignment to that point. The quality score for the best alignment
to any point is equal to the sum of the scoring matrix values of
the matches in that alignment, less the gap creation penalty
multiplied by the number of gaps in that alignment, less the gap
extension penalty multiplied by the total length of all gaps in
that alignment. The gap creation and gap extension penalties are
set by the user. If the best path to any point has a negative
value, a zero is put in that position.
[0205] After the path matrix is complete, the highest value on the
surface of comparison represents the end of the best region of
similarity between the sequences. The best path from this highest
value backwards to the point where the values revert to zero is the
alignment shown by BestFit. This alignment is the best segment of
similarity between the two sequences.
[0206] Additional algorithms for sequence analysis are known in the
art and include ADVANCE and ADAM as described in Torellis and
Robotti (1994) Comput. Appl. Biosci., 10:3-5; and FASTA described
in Pearson and Lipman (1988) Proc. Natl. Acad. Sci. 85:2444-8.
Within FASTA, ktup is a control option that sets the sensitivity
and speed of the search. If ktup=2, similar regions in the two
sequences being compared are found by looking at pairs of aligned
residues; if ktup=1, single aligned amino acids are examined. ktup
can be set to 2 or 1 for protein sequences, or from 1 to 6 for DNA
sequences. The default if ktup is not specified is 2 for proteins
and 6 for DNA. For a further description of FASTA parameters, see
http://bioweb.pasteur.fr/docs/man/man/fasta.1.html#sect2, the
contents of which are incorporated herein by reference.
[0207] The percent identity between two sequences can be determined
using techniques similar to those described above, with or without
allowing gaps. In calculating percent identity, typically exact
matches are counted.
[0208] As used herein, the phrase "allelic variant" refers to a
nucleotide sequence which occurs at a given locus or to a
polypeptide encoded by the nucleotide sequence. For example, the
TANGO 273 gene exhibits significant homology with a portion of
chromosome 7 between chromosomal markers D7S2467 and D7S2552.
Allelic variants of any of this gene can be identified by
sequencing the corresponding chromosomal portion at the indicated
location in multiple individuals.
[0209] As used herein, the terms "gene" and "recombinant gene"
refer to nucleic acid molecules comprising an open reading frame
encoding a polypeptide of the invention. Such natural allelic
variations can typically result in 1-5% variance in the nucleotide
sequence of a given gene. Alternative alleles can be identified by
sequencing the gene of interest in a number of different
individuals. This can be readily carried out by using hybridization
probes to identify the same genetic locus in a variety of
individuals. Any and all such nucleotide variations and resulting
amino acid polymorphisms or variations that are the result of
natural allelic variation and that do not alter the functional
activity are intended to be within the scope of the invention.
[0210] Moreover, nucleic acid molecules encoding proteins of the
invention from other species (homologs), which have a nucleotide
sequence which differs from that of the human proteins described
herein are within the scope of the invention. Nucleic acid
molecules corresponding to natural allelic variants and homologs of
a cDNA of the invention can be isolated based on their identity to
human nucleic acid molecules using the human cDNAs, or a portion
thereof, as a hybridization probe according to standard
hybridization techniques under stringent hybridization conditions.
For example, a cDNA encoding a soluble form of a membrane-bound
protein of the invention can be isolated based on its hybridization
with a nucleic acid molecule encoding all or part of the
membrane-bound form. Likewise, a cDNA encoding a membrane-bound
form can be isolated based on its hybridization with a nucleic acid
molecule encoding all or part of the soluble form.
[0211] Accordingly, in another embodiment, an isolated nucleic acid
molecule of the invention is at least 15 (25, 40, 60, 80, 100, 150,
200, 250, 300, 350, 400, 450, 500, 600, 700, 800, 900, 1000, 1200,
1400, 1600, 1800, 2000, 2200, 2400, 2600, 2800, 3000, or 3500 or
more) nucleotides in length and hybridizes under stringent
conditions to the nucleic acid molecule comprising the nucleotide
sequence, preferably the coding sequence, of any of SEQ ID NOs: 1,
2, 11, 12, 21, 22, 31, 32, 41, 42, 51, 52, 61, 62, 71, 72, 81, 82,
and the nucleotide sequence of any of the clones deposited as
ATCC.RTM. Accession numbers 207185, 207221, PTA-147, PTA-425, and
PTA-424, or a complement thereof. As used herein, the term
"hybridizes under stringent conditions" is intended to describe
conditions for hybridization and washing under which nucleotide
sequences at least 60% (65%, 70%, preferably 75%) identical to each
other typically remain hybridized with each other. Such stringent
conditions are known to those skilled in the art and can be found
in Current Protocols in Molecular Biology, John Wiley & Sons,
N.Y. (1989), 6.3.1-6.3.6. A example of stringent hybridization
conditions are hybridization in 6.times. sodium chloride/sodium
citrate (SSC) at about 45.degree. C., followed by one or more
washes in 0.2.times.SSC, 0.1% SDS at 50-65.degree. C. Preferably,
an isolated nucleic acid molecule of the invention that hybridizes
under stringent conditions to the sequence of any of SEQ ID NOs: 1,
2, 11, 12, 21, 22, 31, 32, 41, 42, 51, 52, 61, 62, 71, 72, 81, 82,
and the nucleotide sequence of any of the clones deposited as
ATCC.RTM. Accession numbers 207185, 207221, PTA-147, PTA-425, and
PTA-424, or a complement thereof, corresponds to a
naturally-occurring nucleic acid molecule. As used herein, a
"naturally-occurring" nucleic acid molecule refers to an RNA or DNA
molecule having a nucleotide sequence that occurs in nature (e.g.,
encodes a natural protein).
[0212] In addition to naturally-occurring allelic variants of a
nucleic acid molecule of the invention sequence that can exist in
the population, the skilled artisan will further appreciate that
changes can be introduced by mutation thereby leading to changes in
the amino acid sequence of the encoded protein, without altering
the biological activity of the protein. For example, one can make
nucleotide substitutions leading to amino acid substitutions at
"non-essential" amino acid residues. A "non-essential" amino acid
residue is a residue that can be altered from the wild-type
sequence without altering the biological activity, whereas an
"essential" amino acid residue is required for biological activity.
For example, amino acid residues that are not conserved or only
semi-conserved among homologs of various species may be
non-essential for activity and thus would be likely targets for
alteration. Alternatively, amino acid residues that are conserved
among the homologs of various species (e.g., murine and human) may
be essential for activity and thus would not be likely targets for
alteration.
[0213] Accordingly, another aspect of the invention pertains to
nucleic acid molecules encoding a polypeptide of the invention that
contain changes in amino acid residues that are not essential for
activity. Such polypeptides differ in amino acid sequence from any
of SEQ ID NOs: 3-8, 13-18, 23-28, 33-38, 43, 53-55, 63-65, 73, and
83-85, yet retain biological activity. In one embodiment, the
isolated nucleic acid molecule includes a nucleotide sequence
encoding a protein that includes an amino acid sequence that is at
least about 40% identical, 50%, 60%, 70%, 80%, 90%, 95%, or 98%
identical to the amino acid sequence of any of SEQ ID NOs: 3-8,
13-18, 23-28, 33-38, 43, 53-55, 63-65, 73, and 83-85, or the amino
acid sequence encoded by the nucleotide sequence of any of the
clones deposited as ATCC.RTM. Accession numbers 207185, 207221,
PTA-147, PTA-425, and PTA-424.
[0214] An isolated nucleic acid molecule encoding a variant protein
can be created by introducing one or more nucleotide substitutions,
additions or deletions into the nucleotide sequence of any of SEQ
ID NOs: 1, 2, 11, 12, 21, 22, 31, 32, 41, 42, 51, 52, 61, 62, 71,
72, 81, 82, and the nucleotide sequence of any of the clones
deposited as ATCC.RTM. Accession numbers 207185, 207221, PTA-147,
PTA-425, and PTA-424, such that one or more amino acid residue
substitutions, additions or deletions are introduced into the
encoded protein. Mutations can be introduced by standard
techniques, such as site-directed mutagenesis and PCR-mediated
mutagenesis. Preferably, conservative amino acid substitutions are
made at one or more predicted non-essential amino acid residues. A
"conservative amino acid substitution" is one in which the amino
acid residue is replaced with an amino acid residue having a
similar side chain. Families of amino acid residues having similar
side chains have been defined in the art. These families include
amino acids with basic side chains (e.g., lysine, arginine,
histidine), acidic side chains (e.g., aspartic acid, glutamic
acid), uncharged polar side chains (e.g., glycine, asparagine,
glutamine, serine, threonine, tyrosine, cysteine), non-polar side
chains (e.g., alanine, valine, leucine, isoleucine, proline,
phenylalanine, methionine, tryptophan), beta-branched side chains
(e.g., threonine, valine, isoleucine) and aromatic side chains
(e.g., tyrosine, phenylalanine, tryptophan, histidine).
Alternatively, mutations can be introduced randomly along all or
part of the coding sequence, such as by saturation mutagenesis, and
the resultant mutants can be screened for biological activity to
identify mutants that retain activity. Following mutagenesis, the
encoded protein can be expressed recombinantly and the activity of
the protein can be determined.
[0215] In one embodiment, a mutant polypeptide that is a variant of
a polypeptide of the invention can be assayed for: (1) the ability
to form protein:protein interactions with the polypeptide of the
invention; (2) the ability to bind a ligand of the polypeptide of
the invention; (3) the ability to bind with a modulator or
substrate of the polypeptide of the invention; or (4) the ability
to modulate a physiological activity of the protein, such as one of
those disclosed herein.
[0216] The present invention encompasses antisense nucleic acid
molecules, i.e., molecules which are complementary to a sense
nucleic acid encoding a polypeptide of the invention, e.g.,
complementary to the coding strand of a double-stranded cDNA
molecule or complementary to an mRNA sequence. Accordingly, an
antisense nucleic acid can hydrogen bond to a sense nucleic acid.
The antisense nucleic acid can be complementary to an entire coding
strand, or to only a portion thereof, e.g., all or part of the
protein coding region (or open reading frame). An antisense nucleic
acid molecule can be antisense to all or part of a non-coding
region of the coding strand of a nucleotide sequence encoding a
polypeptide of the invention. The non-coding regions ("5' and 3'
non-translated regions") are the 5' and 3' sequences which flank
the coding region and are not translated into amino acids.
[0217] An antisense oligonucleotide can be, for example, about 5,
10, 15, 20, 25, 30, 35, 40, 45, or 50 or more nucleotides in
length. An antisense nucleic acid of the invention can be
constructed using chemical synthesis and enzymatic ligation
reactions using procedures known in the art. For example, an
antisense nucleic acid (e.g., an antisense oligonucleotide) can be
chemically synthesized using naturally occurring nucleotides or
variously modified nucleotides designed to increase the biological
stability of the molecules or to increase the physical stability of
the duplex formed between the antisense and sense nucleic acids,
e.g., phosphorothioate derivatives and acridine substituted
nucleotides can be used. Examples of modified nucleotides which can
be used to generate the antisense nucleic acid include
5-fluorouracil, 5-bromouracil, 5-chlorouracil, 5-iodouracil,
hypoxanthine, xanthine, 4-acetylcytosine, 5-(carboxyhydroxylmethyl)
uracil, 5-carboxymethylaminomethyl-2-thiouridine,
5-carboxymethylaminomethyluracil, dihydrouracil,
beta-D-galactosylqueosine, inosine, N6-isopentenyladenine,
1-methylguanine, 1-methylinosine, 2,2-dimethylguanine,
2-methyladenine, 2-methylguanine, 3-methylcytosine,
5-methylcytosine, N6-adenine, 7-methylguanine, 5-methylaminomethyl
uracil, 5-methoxyaminomethyl-2-thiouracil, beta-D-mannosylqueosine,
5'-methoxycarboxymethyluracil, 5-methoxyuracil,
2-methylthio-N6-isopentenyladenine, uracil-5-oxyacetic acid (v),
wybutoxosine, pseudouracil, queosine, 2-thiocytosine,
5-methyl-2-thiouracil, 2-thiouracil, 4-thiouracil, 5-methyluracil,
uracil-5-oxyacetic acid methylester, uracil-5-oxyacetic acid (v),
5-methyl-2-thiouracil, 3-(3-amino-3-N-2-carboxypropyl) uracil,
(acp3)w, and 2,6-diaminopurine. Alternatively, the antisense
nucleic acid can be produced biologically using an expression
vector into which a nucleic acid has been sub-cloned in an
antisense orientation (i.e., RNA transcribed from the inserted
nucleic acid will be of an antisense orientation to a target
nucleic acid of interest, described further in the following
subsection).
[0218] The antisense nucleic acid molecules of the invention are
typically administered to a subject or generated in situ such that
they hybridize with or bind with cellular mRNA and/or genomic DNA
encoding a selected polypeptide of the invention to thereby inhibit
expression, e.g., by inhibiting transcription and/or translation.
The hybridization can be by conventional nucleotide complementarity
to form a stable duplex, or, for example, in the case of an
antisense nucleic acid molecule which binds with DNA duplexes,
through specific interactions in the major groove of the double
helix. An example of a route of administration of antisense nucleic
acid molecules of the invention includes direct injection at a
tissue site. Alternatively, antisense nucleic acid molecules can be
modified to target selected cells and then administered
systemically. For example, for systemic administration, antisense
molecules can be modified such that they specifically bind with
receptors or antigens expressed on a selected cell surface, e.g.,
by linking the antisense nucleic acid molecules to peptides or
antibodies which bind with cell surface receptors or antigens. The
antisense nucleic acid molecules can also be delivered to cells
using the vectors described herein. To achieve sufficient
intracellular concentrations of the antisense molecules, vector
constructs in which the antisense nucleic acid molecule is placed
under the control of a strong pol II or pol III promoter are
preferred.
[0219] An antisense nucleic acid molecule of the invention can be
an alpha-anomeric nucleic acid molecule. An alpha-anomeric nucleic
acid molecule forms specific double-stranded hybrids with
complementary RNA in which the strands run parallel to each other
(Gaultier et al. (1987) Nucleic Acids Res. 15:6625-6641). The
antisense nucleic acid molecule can also comprise a
2'-o-methylribonucleotide (Inoue et al. (1987) Nucleic Acids Res.
15:6131-6148) or a chimeric RNA-DNA analogue (Inoue et al. (1987)
FEBS Lett. 215:327-330).
[0220] The invention also encompasses ribozymes. Ribozymes are
catalytic RNA molecules which are capable of cleaving a
single-stranded nucleic acid, such as an mRNA, to which they have a
complementary region. Thus, ribozymes (e.g., hammerhead ribozymes
as described in Haselhoff and Gerlach (1988) Nature 334:585-591)
can be used to catalytically cleave mRNA transcripts to thereby
inhibit translation of the protein encoded by the mRNA. A ribozyme
having specificity for a nucleic acid molecule encoding a
polypeptide of the invention can be designed based upon the
nucleotide sequence of a cDNA disclosed herein. For example, a
derivative of a Tetrahymena L-19 IVS RNA can be constructed in
which the nucleotide sequence of the ribozyme active site is
complementary to the nucleotide sequence to be cleaved, as
described in Cech et al. U.S. Pat. No. 4,987,071; and Cech et al.
U.S. Pat. No. 5,116,742. Alternatively, an mRNA encoding a
polypeptide of the invention can be used to select a catalytic RNA
having a specific ribonuclease activity from a pool of RNA
molecules. See, e.g., Bartel and Szostak (1993) Science
261:1411-1418.
[0221] The invention includes nucleic acid molecules which form
triple helical structures. For example, expression of a polypeptide
of the invention can be inhibited by targeting nucleotide sequences
complementary to the regulatory region of the gene encoding the
polypeptide (e.g., the promoter and/or enhancer) to form triple
helical structures that prevent transcription of the gene in target
cells. See generally Helene (1991) Anticancer Drug Des.
6(6):569-84; Helene (1992) Ann. N.Y. Acad. Sci. 660:27-36; and
Maher (1992) Bioassays 14(12):807-15. "Expression" of a
polypeptide, as used herein, refers individually and collectively
to the processes of transcription of DNA to generate an RNA
transcript and translation of an RNA to generate the
polypeptide.
[0222] In various embodiments, the nucleic acid molecules of the
invention can be modified at the base moiety, sugar moiety, or
phosphate backbone to improve, e.g., the stability, hybridization,
or solubility of the molecule. For example, the deoxyribose
phosphate backbone of the nucleic acids can be modified to generate
peptide nucleic acids (see Hyrup et al. (1996) Bioorganic &
Medicinal Chemistry 4(1): 5-23). As used herein, the terms "peptide
nucleic acids" or "PNAs" refer to nucleic acid mimics, e.g., DNA
mimics, in which the deoxyribose phosphate backbone is replaced by
a pseudopeptide backbone and only the four natural nucleobases are
retained. The neutral backbone of PNAs has been shown to allow
specific hybridization with DNA and RNA under conditions of low
ionic strength. Synthesis of PNA oligomers can be performed using
standard solid phase peptide synthesis protocols such as those
described in Hyrup et al. (1996), supra; Perry-O'Keefe et al.
(1996) Proc. Natl. Acad. Sci. USA 93: 14670-675.
[0223] PNAs can be used in therapeutic and diagnostic applications.
For example, PNAs can be used as antisense or anti-gene agents for
sequence-specific modulation of gene expression by, e.g., inducing
arrest of transcription or translation or by inhibiting
replication. PNAs can also be used, e.g., in the analysis of single
base pair mutations in a gene by, e.g., PNA directed PCR clamping;
as artificial restriction enzymes when used in combination with
other enzymes, e.g., S1 nucleases (Hyrup (1996), supra; or as
probes or primers for DNA sequence and hybridization (Hyrup (1996),
supra; Perry-O'Keefe et al. (1996) Proc. Natl. Acad. Sci. USA 93:
14670-675).
[0224] In another embodiment, PNAs can be modified, e.g., to
enhance their stability or cellular uptake, by attaching lipophilic
or other helper groups to PNA, by formation of PNA-DNA chimeras, or
by use of liposomes or other techniques of drug delivery known in
the art. For example, PNA-DNA chimeras can be generated which can
combine the advantageous properties of PNA and DNA. Such chimeras
allow DNA recognition enzymes, e.g., RNase H and DNA polymerases,
to interact with the DNA portion while the PNA portion provides
high binding affinity and specificity. PNA-DNA chimeras can be
linked using linkers of appropriate lengths selected in terms of
base stacking, number of bonds between the nucleobases, and
orientation (Hyrup (1996), supra). The synthesis of PNA-DNA
chimeras can be performed as described in Hyrup (1996), supra, and
Finn et al. (1996) Nucleic Acids Res. 24(17):3357-63. For example,
a DNA chain can be synthesized on a solid support using standard
phosphoramidite coupling chemistry and modified nucleoside analogs.
Compounds such as 5'-(4-methoxytrityl)amino-5'-deoxy-thymidine
phosphoramidite can be used as a link between the PNA and the 5'
end of DNA (Mag et al. (1989) Nucleic Acids Res. 17:5973-88). PNA
monomers are then coupled in a step-wise manner to produce a
chimeric molecule with a 5' PNA segment and a 3' DNA segment (Finn
et al. (1996) Nucleic Acids Res. 24(17):3357-63). Alternatively,
chimeric molecules can be synthesized with a 5' DNA segment and a
3' PNA segment (Peterser et al. (1975) Bioorganic Med. Chem. Lett.
5:1119-11124).
[0225] In other embodiments, the oligonucleotide can include other
appended groups such as peptides (e.g., for targeting host cell
receptors in vivo), or agents facilitating transport across the
cell membrane (see, e.g., Letsinger et al. (1989) Proc. Natl. Acad.
Sci. USA 86:6553-6556; Lemaitre et al. (1987) Proc. Natl. Acad.
Sci. USA 84:648-652; PCT Publication No. WO 88/09810) or the
blood-brain barrier (see, e.g., PCT Publication No. WO 89/10134).
In addition, oligonucleotides can be modified with
hybridization-triggered cleavage agents (see, e.g., Krol et al.
(1988) Bio/Techniques 6:958-976) or intercalating agents (see,
e.g., Zon (1988) Pharm. Res. 5:539-549). To this end, the
oligonucleotide can be conjugated with another molecule, e.g., a
peptide, hybridization triggered cross-linking agent, transport
agent, hybridization-triggered cleavage agent, etc.
II. Isolated Proteins and Antibodies
[0226] One aspect of the invention pertains to isolated proteins,
and biologically active portions thereof, as well as polypeptide
fragments suitable for use as immunogens to generate antibodies
directed against a polypeptide of the invention. In one embodiment,
the native polypeptide is isolated from cells or tissue sources by
an appropriate purification scheme using standard protein
purification techniques. In another embodiment, polypeptides of the
invention are produced by recombinant DNA techniques. As an
alternative to recombinant expression, a polypeptide of the
invention can be synthesized chemically using standard peptide
synthesis techniques.
[0227] An "isolated" or "purified" protein or biologically active
portion thereof is substantially free of cellular material or other
contaminating proteins from the cell or tissue source from which
the protein is derived, or substantially free of chemical
precursors or other chemicals, when chemically synthesized. The
language "substantially free of cellular material" includes
preparations of protein in which the protein is separated from
cellular components of the cells from which it is isolated or
recombinantly produced. Thus, protein that is substantially free of
cellular material includes preparations of protein having less than
about 30%, 20%, 10%, or 5% (by dry weight) of heterologous protein
(also referred to herein as a "contaminating protein"). When the
protein or biologically active portion thereof is recombinantly
produced, it is also preferably substantially free of culture
medium, i.e., culture medium represents less than about 20%, 10%,
or 5% of the volume of the protein preparation. When the protein is
produced by chemical synthesis, it is preferably substantially free
of chemical precursors or other chemicals, i.e., it is separated
from chemical precursors or other chemicals which are involved in
the synthesis of the protein. Accordingly such preparations of the
protein have less than about 30%, 20%, 10%, 5% (by dry weight) of
chemical precursors or compounds other than the polypeptide of
interest.
[0228] Biologically active portions of a polypeptide of the
invention include polypeptide regions having an amino acid sequence
sufficiently identical to or derived from the amino acid sequence
of the protein (e.g., the amino acid sequence shown in any of SEQ
ID NOs: 3-8, 13-18, 23-28, 33-38, 43, 53-55, 63-65, 73, and 83-85,
or the amino acid sequence encoded by the nucleotide sequence of
any of the clones deposited as ATCC.RTM. Accession numbers 207185,
207221, PTA-147, PTA-425, and PTA-424), which include fewer amino
acids than the full length protein, and exhibit at least one
activity of the corresponding full-length protein. Typically,
biologically active portions comprise a domain or motif with at
least one activity of the corresponding protein. A biologically
active portion of a protein of the invention can be a polypeptide
which is, for example, 10, 25, 50, 100 or more amino acids in
length. Moreover, other biologically active portions, in which
other regions of the protein are deleted, can be prepared by
recombinant techniques and evaluated for one or more of the
functional activities of the native form of a polypeptide of the
invention.
[0229] Examples of polypeptides are those which have the amino acid
sequence of any of SEQ ID NOs: 3-8, 13-18, 23-28, 33-38, 43, 53-55,
63-65, 73, and 83-85 or the amino acid sequence encoded by the
nucleotide sequence of any of the clones deposited as ATCC.RTM.
Accession numbers 207185, 207221, PTA-147, PTA-425, and PTA-424.
Other useful proteins are substantially identical (e.g., at least
about 40%, preferably 50%, 60%, 70%, 80%, 90%, 95%, or 99%) to any
of SEQ ID NOs: 3-8, 13-18, 23-28, 33-38, 43, 53-55, 63-65, 73, and
83-85 or the amino acid sequence encoded by the nucleotide sequence
of any of the clones deposited as ATCC.RTM. Accession numbers
207185, 207221, PTA-147, PTA-425, and PTA-424 and retain the
functional activity of the protein of the corresponding
naturally-occurring protein. Such proteins can differ in amino acid
sequence owing, for example, to natural allelic variation or
mutagenesis.
[0230] The invention also provides chimeric or fusion proteins. As
used herein, a "chimeric protein" or "fusion protein" comprises all
or part (preferably biologically active) of a polypeptide of the
invention operably linked with a heterologous polypeptide (i.e., a
polypeptide other than the same polypeptide of the invention).
Within the fusion protein, the term "operably linked" is intended
to indicate that the polypeptide of the invention and the
heterologous polypeptide are fused in-frame with each other. The
heterologous polypeptide can be fused with the amino-terminus or
the carboxyl-terminus of the polypeptide of the invention.
[0231] One useful fusion protein is a GST fusion protein in which
the polypeptide of the invention is fused with the carboxyl
terminus of GST sequences. Such fusion proteins can facilitate
purification of a recombinant polypeptide of the invention.
[0232] In another embodiment, the fusion protein contains a
heterologous signal sequence at its amino terminus. For example,
the native signal sequence of a polypeptide of the invention can be
removed and replaced with a signal sequence from another protein.
For example, the gp67 secretory sequence of the baculovirus
envelope protein can be used as a heterologous signal sequence
(Current Protocols in Molecular Biology, Ausubel et al., eds., John
Wiley & Sons, 1992). Other examples of eukaryotic heterologous
signal sequences include the secretory sequences of melittin and
human placental alkaline phosphatase (Stratagene; La Jolla,
Calif.). In yet another example, useful prokaryotic heterologous
signal sequences include the phoA secretory signal (Sambrook et
al., supra) and the protein A secretory signal (Pharmacia Biotech;
Piscataway, N.J.).
[0233] In yet another embodiment, the fusion protein is an
immunoglobulin fusion protein in which all or part of a polypeptide
of the invention is fused with sequences derived from a member of
the immunoglobulin protein family. The immunoglobulin fusion
proteins of the invention can be incorporated into pharmaceutical
compositions and administered to a subject to inhibit an
interaction between a ligand (soluble or membrane-bound) and a
protein on the surface of a cell (receptor), to thereby suppress
signal transduction in vivo. The immunoglobulin fusion protein can
be used to affect the bioavailability of a cognate ligand of a
polypeptide of the invention. Inhibition of ligand/receptor
interaction can be useful therapeutically, both for treating
proliferative and differentiative disorders and for modulating
(e.g., promoting or inhibiting) cell survival. Moreover, the
immunoglobulin fusion proteins of the invention can be used as
immunogens to produce antibodies directed against a polypeptide of
the invention in a subject, to purify ligands and in screening
assays to identify molecules which inhibit the interaction of
receptors with ligands. The immunoglobulin fusion protein can, for
example, comprise a portion of a polypeptide of the invention fused
with the amino-terminus or the carboxyl-terminus of an
immunoglobulin constant region, as disclosed in U.S. Pat. No.
5,714,147, U.S. Pat. No. 5,116,964, U.S. Pat. No. 5,514,582, and
U.S. Pat. No. 5,455,165.
[0234] Chimeric and fusion proteins of the invention can be
produced by standard recombinant DNA techniques. In another
embodiment, the fusion gene can be synthesized by conventional
techniques including automated DNA synthesizers. Alternatively, PCR
amplification of gene fragments can be performed using anchor
primers which give rise to complementary overhangs between two
consecutive gene fragments and which can subsequently be annealed
and re-amplified to generate a chimeric gene sequence (see, e.g.,
Ausubel et al., supra). Moreover, many expression vectors are
commercially available that already encode a fusion moiety (e.g., a
GST polypeptide). A nucleic acid encoding a polypeptide of the
invention can be cloned into such an expression vector such that
the fusion moiety is linked in-frame to the polypeptide of the
invention.
[0235] A signal sequence of a polypeptide of the invention (e.g.,
the signal sequence in any of SEQ ID NOs: 3, 13, 23, 33, 43, 53,
63, 73, and 83) can be used to facilitate secretion and isolation
of the secreted protein or another protein of interest. Signal
sequences are typically characterized by a core of hydrophobic
amino acids which are generally cleaved from the mature protein
during secretion in one or more cleavage events. Such signal
peptides contain processing sites that allow cleavage of the signal
sequence from the mature proteins as they pass through the
secretory pathway. Thus, the invention pertains to the described
polypeptides having a signal sequence, as well as to the signal
sequence itself and to the polypeptide in the absence of the signal
sequence (i.e., the cleavage products). In one embodiment, a
nucleic acid sequence encoding a signal sequence of the invention
can be operably linked in an expression vector with a protein of
interest, such as a protein which is ordinarily not secreted or is
otherwise difficult to isolate. The signal sequence directs
secretion of the protein, such as from a eukaryotic host into which
the expression vector is transformed, and the signal sequence is
subsequently or concurrently cleaved. The protein can then be
readily purified from the extracellular medium by art recognized
methods. Alternatively, the signal sequence can be linked with the
protein of interest using a sequence which facilitates
purification, such as with a GST domain.
[0236] In another embodiment, the signal sequences of the present
invention can be used to identify regulatory sequences, e.g.,
promoters, enhancers, repressors. Since signal sequences are the
most amino-terminal sequences of a peptide, the nucleic acids which
flank the signal sequence on its amino-terminal side are likely
regulatory sequences which affect transcription. Thus, a nucleotide
sequence which encodes all or a portion of a signal sequence can be
used as a probe to identify and isolate signal sequences and their
flanking regions, and these flanking regions can be studied to
identify regulatory elements therein.
[0237] The present invention also pertains to variants of the
polypeptides of the invention. Such variants have an altered amino
acid sequence which can function as either agonists (mimetics) or
as antagonists. Variants can be generated by mutagenesis, e.g.,
discrete point mutation or truncation. An agonist can retain
substantially the same, or a subset, of the biological activities
of the naturally occurring form of the protein. An antagonist of a
protein can inhibit one or more of the activities of the naturally
occurring form of the protein by, for example, competitively
binding with a downstream or upstream member of a cellular
signaling cascade which includes the protein of interest. Thus,
specific biological effects can be elicited by treatment with a
variant of limited function. Treatment of a subject with a variant
having a subset of the biological activities of the naturally
occurring form of the protein can have fewer side effects in a
subject, relative to treatment with the naturally occurring form of
the protein.
[0238] Variants of a protein of the invention which function as
either agonists (e.g., mimetics) or as antagonists can be
identified by screening combinatorial libraries of mutants, e.g.,
truncation mutants, of the protein of the invention for agonist or
antagonist activity. In one embodiment, a variegated library of
variants is generated by combinatorial mutagenesis at the nucleic
acid level and is encoded by a variegated gene library. A
variegated library of variants can be produced by, for example,
enzymatically ligating a mixture of synthetic oligonucleotides into
gene sequences such that a degenerate set of potential protein
sequences can be expressed as individual polypeptides, or
alternatively, as a set of larger fusion proteins (e.g., for phage
display). There are a variety of methods which can be used to
produce libraries of potential variants of the polypeptides of the
invention from a degenerate oligonucleotide sequence. Methods for
synthesizing degenerate oligonucleotides are known in the art (see,
e.g., Narang (1983) Tetrahedron 39:3; Itakura et al. (1984) Annu.
Rev. Biochem. 53:323; Itakura et al. (1984) Science 198:1056; Ike
et al. (1983) Nucleic Acid Res. 11:477).
[0239] In addition, libraries of fragments of the coding sequence
of a polypeptide of the invention can be used to generate a
variegated population of polypeptides for screening and subsequent
selection of variants. For example, a library of coding sequence
fragments can be generated by treating a double stranded PCR
fragment of the coding sequence of interest with a nuclease under
conditions wherein nicking occurs only about once per molecule,
denaturing the double stranded DNA, re-naturing the DNA to form
double stranded DNA which can include sense/antisense pairs from
different nicked products, removing single stranded portions from
reformed duplexes by treatment with S1 nuclease, and ligating the
resulting fragment library into an expression vector. By this
method, an expression library can be derived which encodes amino
terminal and internal fragments of various sizes of the protein of
interest.
[0240] Several techniques are known in the art for screening gene
products of combinatorial libraries made by point mutations or
truncation, and for screening cDNA libraries for gene products
having a selected property. The most widely used techniques, which
are amenable to high through-put analysis, for screening large gene
libraries typically include cloning the gene library into
replicable expression vectors, transforming appropriate cells with
the resulting library of vectors, and expressing the combinatorial
genes under conditions in which detection of a desired activity
facilitates isolation of the vector encoding the gene whose product
was detected. Recursive ensemble mutagenesis (REM), a technique
which enhances the frequency of functional mutants in the
libraries, can be used in combination with the screening assays to
identify variants of a protein of the invention (Arkin and Yourvan
(1992) Proc. Natl. Acad. Sci. USA 89:7811-7815; Delgrave et al.
(1993) Protein Engineering 6(3):327-331).
[0241] An isolated polypeptide of the invention, or a fragment
thereof, can be used as an immunogen to generate antibodies using
standard techniques for polyclonal and monoclonal antibody
preparation. The full-length polypeptide or protein can be used or,
alternatively, the invention provides antigenic peptide fragments
for use as immunogens. The antigenic peptide of a protein of the
invention comprises at least 10 (preferably 12, 15, 20, or 30 or
more) amino acid residues of the amino acid sequence of any of SEQ
ID NOs: 3-8, 13-18, 23-28, 33-38, 43, 53-55, 63-65, 73, and 83-85
or the amino acid sequence encoded by the nucleotide sequence of
any of the clones deposited as ATCC.RTM. Accession numbers 207185,
207221, PTA-147, PTA-425, and PTA-424, and encompasses an epitope
of the protein such that an antibody raised against the peptide
forms a specific immune complex with the protein.
[0242] Examples of epitopes encompassed by the antigenic peptide
are regions that are located on the surface of the protein, e.g.,
hydrophilic regions. FIGS. 1I, 1J, 2F, 3F, 4D, 4G, and 5C are
hydrophobicity plots of proteins of the invention. These plots or
similar analyses can be used to identify hydrophilic regions.
[0243] An immunogen typically is used to prepare antibodies by
immunizing a suitable (i.e., immunocompetent) subject such as a
rabbit, goat, mouse, or other mammal or vertebrate. An appropriate
immunogenic preparation can contain, for example,
recombinantly-expressed or chemically-synthesized polypeptide. The
preparation can further include an adjuvant, such as Freund's
complete or incomplete adjuvant, or a similar immunostimulatory
agent.
[0244] Accordingly, another aspect of the invention pertains to
antibodies directed against a polypeptide of the invention. The
terms "antibody" and "antibody substance" as used interchangeably
herein refer to immunoglobulin molecules and immunologically active
portions of immunoglobulin molecules, i.e., molecules that contain
an antigen binding site which specifically binds an antigen, such
as a polypeptide of the invention. A molecule which specifically
binds with a given polypeptide of the invention is a molecule which
binds the polypeptide, but does not substantially bind other
molecules in a sample, e.g., a biological sample, which naturally
contains the polypeptide. Examples of immunologically active
portions of immunoglobulin molecules include F(ab) and F(ab')2
fragments which can be generated by treating the antibody with an
enzyme such as papain or pepsin, respectively. The invention
provides polyclonal and monoclonal antibodies. The term "monoclonal
antibody" or "monoclonal antibody composition", as used herein,
refers to a population of antibody molecules that contain only one
species of an antigen binding site capable of immunoreacting with a
particular epitope.
[0245] Polyclonal antibodies can be prepared as described above by
immunizing a suitable subject with a polypeptide of the invention
as an immunogen. The antibody titer in the immunized subject can be
monitored over time by standard techniques, such as with an enzyme
linked immunosorbent assay (ELISA) using immobilized polypeptide.
If desired, the antibody molecules can be harvested or isolated
from the subject (e.g., from the blood or serum of the subject) and
further purified by well-known techniques, such as protein A
chromatography to obtain the IgG fraction. At an appropriate time
after immunization, e.g., when the specific antibody titers are
highest, antibody-producing cells can be obtained from the subject
and used to prepare monoclonal antibodies by standard techniques,
such as the hybridoma technique originally described by Kohler and
Milstein (1975) Nature 256:495-497, the human B cell hybridoma
technique (Kozbor et al. (1983) Immunol. Today 4:72), the
EBV-hybridoma technique (Cole et al. (1985), Monoclonal Antibodies
and Cancer Therapy, Alan R. Liss, Inc., pp. 77-96) or trioma
techniques. The technology for producing hybridomas is well known
(see generally Current Protocols in Immunology (1994) Coligan et
al. (Eds.) John Wiley & Sons, Inc., New York, N.Y.). Hybridoma
cells producing a monoclonal antibody of the invention are detected
by screening the hybridoma culture supernatants for antibodies that
bind the polypeptide of interest, e.g., using a standard ELISA
assay.
[0246] Alternative to preparing monoclonal antibody-secreting
hybridomas, a monoclonal antibody directed against a polypeptide of
the invention can be identified and isolated by screening a
recombinant combinatorial immunoglobulin library (e.g., an antibody
phage display library) with the polypeptide of interest. Kits for
generating and screening phage display libraries are commercially
available (e.g., the Pharmacia Recombinant Phage Antibody System,
Catalog No. 27-9400-01; and the Stratagene SURFZAP.TM. Phage
Display Kit, Catalog No. 240612). Additionally, examples of methods
and reagents particularly amenable for use in generating and
screening antibody display library can be found in, for example,
U.S. Pat. No. 5,223,409; PCT Publication No. WO 92/18619; PCT
Publication No. WO 91/17271; PCT Publication No. WO 92/20791; PCT
Publication No. WO 92/15679; PCT Publication No. WO 93/01288; PCT
Publication No. WO 92/01047; PCT Publication No. WO 92/09690; PCT
Publication No. WO 90/02809; Fuchs et al. (1991) Bio/Technology
9:1370-1372; Hay et al. (1992) Hum. Antibod. Hybridomas 3:81-85;
Huse et al. (1989) Science 246:1275-1281; Griffiths et al. (1993)
EMBO J. 12:725-734.
[0247] Recombinant antibodies, such as chimeric and humanized
monoclonal antibodies, comprising both human and non-human
portions, which can be made using standard recombinant DNA
techniques, are within the scope of the invention. Such chimeric
and humanized monoclonal antibodies can be produced by recombinant
DNA techniques known in the art, for example using methods
described in PCT Publication No. WO 87/02671; European Patent
Application 184,187; European Patent Application 171,496; European
Patent Application 173,494; PCT Publication No. WO 86/01533; U.S.
Pat. No. 4,816,567; European Patent Application 125,023; Better et
al. (1988) Science 240:1041-1043; Liu et al. (1987) Proc. Natl.
Acad. Sci. USA 84:3439-3443; Liu et al. (1987) J. Immunol.
139:3521-3526; Sun et al. (1987) Proc. Natl. Acad. Sci. USA
84:214-218; Nishimura et al. (1987) Cancer Res. 47:999-1005; Wood
et al. (1985) Nature 314:446-449; and Shaw et al. (1988) J. Natl.
Cancer Inst. 80:1553-1559); Morrison (1985) Science 229:1202-1207;
Oi et al. (1986) Bio/Techniques 4:214; U.S. Pat. No. 5,225,539;
Jones et al. (1986) Nature 321:552-525; Verhoeyan et al. (1988)
Science 239:1534; and Beidler et al. (1988) J. Immunol.
141:4053-4060.
[0248] Completely human antibodies are particularly desirable for
therapeutic treatment of human patients. Such antibodies can be
produced using transgenic mice which are incapable of expressing
endogenous immunoglobulin heavy and light chains genes, but which
can express human heavy and light chain genes. The transgenic mice
are immunized in the normal fashion with a selected antigen, e.g.,
all or a portion of a polypeptide of the invention. Monoclonal
antibodies directed against the antigen can be obtained using
conventional hybridoma technology. The human immunoglobulin
transgenes harbored by the transgenic mice rearrange during B cell
differentiation, and subsequently undergo class switching and
somatic mutation. Thus, using such a technique, it is possible to
produce therapeutically useful IgG, IgA, and IgE antibodies. For an
overview of this technology for producing human antibodies, see
Lonberg and Huszar (1995, Int. Rev. Immunol. 13:65-93). For a
detailed discussion of this technology for producing human
antibodies and human monoclonal antibodies and protocols for
producing such antibodies, see, e.g., U.S. Pat. No. 5,625,126; U.S.
Pat. No. 5,633,425; U.S. Pat. No. 5,569,825; U.S. Pat. No.
5,661,016; and U.S. Pat. No. 5,545,806. In addition, companies such
as Abgenix, Inc. (Freemont, Calif.), can be engaged to provide
human antibodies directed against a selected antigen using
technology similar to that described above.
[0249] Completely human antibodies which recognize a selected
epitope can be generated using a technique referred to as "guided
selection." In this approach a selected non-human monoclonal
antibody, e.g., a murine antibody, is used to guide the selection
of a completely human antibody recognizing the same epitope
(Jespers et al. (1994) Bio/technology 12:899-903).
[0250] An antibody directed against a polypeptide of the invention
(e.g., monoclonal antibody) can be used to isolate the polypeptide
by standard techniques, such as affinity chromatography or
immunoprecipitation. Moreover, such an antibody can be used to
detect the protein (e.g., in a cellular lysate or cell supernatant)
in order to evaluate the abundance and pattern of expression of the
polypeptide. The antibodies can also be used diagnostically to
monitor protein levels in tissue as part of a clinical testing
procedure, e.g., to, for example, determine the efficacy of a given
treatment regimen. Detection can be facilitated by coupling the
antibody to a detectable substance. Examples of detectable
substances include various enzymes, prosthetic groups, fluorescent
materials, luminescent materials, bioluminescent materials, and
radioactive materials. Examples of suitable enzymes include
horseradish peroxidase, alkaline phosphatase, beta-galactosidase,
or acetylcholinesterase; examples of suitable prosthetic group
complexes include streptavidin/biotin and avidin/biotin; examples
of suitable fluorescent materials include umbelliferone,
fluorescein, fluorescein isothiocyanate, rhodamine,
dichlorotriazinylamine fluorescein, dansyl chloride or
phycoerythrin; an example of a luminescent material includes
luminol; examples of bioluminescent materials include luciferase,
luciferin, and aequorin, and examples of suitable radioactive
material include .sup.125I, .sup.131I, .sup.35S or .sup.3H.
[0251] An antibody (or fragment thereof) can be conjugated to a
therapeutic moiety such as a cytotoxin, a therapeutic agent, or a
radioactive agent (e.g., a radioactive metal ion). Cytotoxins and
cytotoxic agents include any agent that is detrimental to cells.
Examples of such agents include taxol, cytochalasin B, gramicidin
D, ethidium bromide, emetine, mitomycin, etoposide, tenoposide,
vincristine, vinblastine, colchicin, doxorubicin, daunorubicin,
dihydroxy anthracin dione, mitoxantrone, mithramycin, actinomycin
D, 1-dehydrotestosterone, glucocorticoids, procaine, tetracaine,
lidocaine, propranolol, and puromycin and analogs or homologs
thereof. Therapeutic agents include, but are not limited to,
antimetabolites (e.g., methotrexate, 6-mercaptopurine,
6-thioguanine, cytarabine, and 5-fluorouracil decarbazine),
alkylating agents (e.g., mechlorethamine, thioepa chlorambucil,
melphalan, carmustine (BSNU) and lomustine (CCNU),
cyclophosphamide, busulfan, dibromomannitol, streptozotocin,
mitomycin C, and cis-dichlorodiamine platinum (II) (DDP)
cisplatin), anthracyclines (e.g., daunorubicin {formerly designated
daunomycin} and doxorubicin), antibiotics (e.g., dactinomycin
{formerly designated actinomycin}, bleomycin, mithramycin, and
anthramycin), and anti-mitotic agents (e.g., vincristine and
vinblastine).
[0252] Conjugated antibodies of the invention can be used for
modifying a given biological response, the drug moiety not being
limited to classical chemical therapeutic agents. For example, the
drug moiety can be a protein or polypeptide possessing a desired
biological activity. Such proteins include, for example, toxins
such as abrin, ricin A, Pseudomonas exotoxin, or diphtheria toxin;
proteins such as tumor necrosis factor, alpha-interferon,
beta-interferon, nerve growth factor, platelet derived growth
factor, tissue plasminogen activator; and biological response
modifiers such as lymphokines, interleukin-1, interleukin-2,
interleukin-6, granulocyte macrophage colony stimulating factor,
granulocyte colony stimulating factor, or other growth factors.
[0253] Techniques for conjugating a therapeutic moiety to an
antibody are well known (see, e.g., Amon et al., 1985, "Monoclonal
Antibodies For Immunotargeting Of Drugs In Cancer Therapy", in
Monoclonal Antibodies And Cancer Therapy, Reisfeld et al., Eds.,
Alan R. Liss, Inc. pp. 243-256; Hellstrom et al., 1987, "Antibodies
For Drug Delivery", in Controlled Drug Delivery, 2nd ed., Robinson
et al., Eds., Marcel Dekker, Inc., pp. 623-653; Thorpe, 1985,
"Antibody Carriers Of Cytotoxic Agents In Cancer Therapy: A
Review", in Monoclonal Antibodies '84: Biological And Clinical
Applications, Pinchera et al., Eds., pp. 475-506; "Analysis,
Results, And Future Prospective Of The Therapeutic Use Of
Radiolabeled Antibody In Cancer Therapy", in Monoclonal Antibodies
For Cancer Detection And Therapy, Baldwin et al., Eds., Academic
Press, pp. 303-316, 1985; and Thorpe et al., 1982, Immunol. Rev.,
62:119-158). Alternatively, an antibody can be conjugated to a
second antibody to form an antibody heteroconjugate as described by
Segal in U.S. Pat. No. 4,676,980.
III. Recombinant Expression Vectors and Host Cells
[0254] Another aspect of the invention pertains to vectors,
including expression vectors, containing a nucleic acid encoding a
polypeptide of the invention (or a portion thereof). As used
herein, the term "vector" refers to a nucleic acid molecule capable
of transporting another nucleic acid to which it has been linked.
One type of vector is a "plasmid", which refers to a circular
double stranded DNA loop into which additional DNA segments can be
ligated. Another type of vector is a viral vector, wherein
additional DNA segments can be ligated into the viral genome.
Certain vectors are capable of autonomous replication in a host
cell into which they are introduced (e.g., bacterial vectors having
a bacterial origin of replication and episomal mammalian vectors).
Other vectors (e.g., non-episomal mammalian vectors) are integrated
into the genome of a host cell upon introduction into the host
cell, and thereby are replicated along with the host genome.
Moreover, certain vectors, designated expression vectors, are
capable of directing expression of genes with which they are
operably linked. In general, expression vectors of utility in
recombinant DNA techniques are often in the form of plasmids
(vectors). However, the invention is intended to include such other
forms of expression vectors, such as viral vectors (e.g.,
replication defective retroviruses, adenoviruses and
adeno-associated viruses), which serve equivalent functions.
[0255] The recombinant expression vectors of the invention comprise
a nucleic acid of the invention in a form suitable for expression
of the nucleic acid in a host cell. This means that the recombinant
expression vectors include one or more regulatory sequences,
selected on the basis of the host cells to be used for expression,
which is operably linked with the nucleic acid sequence to be
expressed. Within a recombinant expression vector, "operably
linked" is intended to mean that the nucleotide sequence of
interest is linked with the regulatory sequence(s) in a manner
which allows expression of the nucleotide sequence (e.g., in an in
vitro transcription/translation system or in a host cell when the
vector is introduced into the host cell). The term "regulatory
sequence" is intended to include promoters, enhancers and other
expression control elements (e.g., polyadenylation signals). Such
regulatory sequences are described, for example, in Goeddel, Gene
Expression Technology Methods in Enzymology 185, Academic Press,
San Diego, Calif. (1990). Regulatory sequences include those which
direct constitutive expression of a nucleotide sequence in many
types of host cell and those which direct expression of the
nucleotide sequence only in certain host cells (e.g.,
tissue-specific regulatory sequences). It will be appreciated by
those skilled in the art that the design of the expression vector
can depend on such factors as the choice of the host cell to be
transformed, and the level of expression of protein desired. The
expression vectors of the invention can be introduced into host
cells to thereby produce proteins or peptides, including fusion
proteins or peptides, encoded by nucleic acids as described
herein.
[0256] The recombinant expression vectors of the invention can be
designed for expression of a polypeptide of the invention in
prokaryotic (e.g., E. coli) or eukaryotic cells (e.g., insect cells
(using baculovirus expression vectors), yeast cells or mammalian
cells). Suitable host cells are discussed further in Goeddel,
supra. Alternatively, the recombinant expression vector can be
transcribed and translated in vitro, for example using T7 promoter
regulatory sequences and T7 polymerase.
[0257] Expression of proteins in prokaryotes is most often carried
out in E. coli with vectors containing constitutive or inducible
promoters directing the expression of either fusion or non-fusion
proteins. Fusion vectors add a number of amino acids to a protein
encoded therein, usually to the amino terminus of the recombinant
protein. Such fusion vectors typically serve three purposes: 1) to
increase expression of recombinant protein; 2) to increase the
solubility of the recombinant protein; and 3) to aid in the
purification of the recombinant protein by acting as a ligand in
affinity purification. Often, in fusion expression vectors, a
proteolytic cleavage site is introduced at the junction of the
fusion moiety and the recombinant protein to enable separation of
the recombinant protein from the fusion moiety subsequent to
purification of the fusion protein. Such enzymes, and their cognate
recognition sequences, include Factor Xa, thrombin and
enterokinase. Typical fusion expression vectors include pGEX
(Pharmacia Biotech Inc; Smith and Johnson (1988) Gene 67:31-40),
pMAL (New England Biolabs, Beverly, Mass.) and pRIT5 (Pharmacia,
Piscataway, N.J.) which fuse glutathione S-transferase (GST),
maltose E binding protein, or protein A, respectively, to the
target recombinant protein.
[0258] Examples of suitable inducible non-fusion E. coli expression
vectors include pTrc (Amann et al., (1988) Gene 69:301-315) and pET
11d (Studier et al., Gene Expression Technology: Methods in
Enzymology 185, Academic Press, San Diego, Calif. (1990) 60-89).
Target gene expression from the pTrc vector relies on host RNA
polymerase transcription from a hybrid trp-lac fusion promoter.
Target gene expression from the pET 11d vector relies on
transcription from a T7 gn10-lac fusion promoter mediated by a
co-expressed viral RNA polymerase (T7 gn1). This viral polymerase
is supplied by host strains BL21 (DE3) or HMS174 (DE3) from a
resident lambda prophage harboring a T7 gn1 gene under the
transcriptional control of the lacUV 5 promoter.
[0259] One strategy to maximize recombinant protein expression in
E. coli is to express the protein in a host bacteria having an
impaired capacity to proteolytically cleave the recombinant protein
(Gottesman, Gene Expression Technology: Methods in Enzymology 185,
Academic Press, San Diego, Calif. (1990) 119-128). Another strategy
is to alter the nucleic acid sequence of the nucleic acid to be
inserted into an expression vector such that the individual codons
for each amino acid are those preferentially used in E. coli (Wada
et al. (1992) Nucleic Acids Res. 20:2111-2118). Such alteration of
nucleic acid sequences of the invention can be performed by
standard DNA synthesis techniques.
[0260] In another embodiment, the expression vector is a yeast
expression vector. Examples of vectors for expression in yeast S.
cerevisiae include pYepSec1 (Baldari et al. (1987) EMBO J.
6:229-234), pMFa (Kurjan and Herskowitz, (1982) Cell 30:933-943),
pJRY88 (Schultz et al. (1987) Gene 54:113-123), pYES2 (Invitrogen
Corporation, San Diego, Calif.), and pPicZ (Invitrogen Corp, San
Diego, Calif.).
[0261] Alternatively, the expression vector is a baculovirus
expression vector. Baculovirus vectors available for expression of
proteins in cultured insect cells (e.g., Sf 9 cells) include the
pAc series (Smith et al. (1983) Mol. Cell. Biol. 3:2156-2165) and
the pVL series (Lucklow and Summers (1989) Virology 170:31-39).
[0262] In yet another embodiment, a nucleic acid of the invention
is expressed in mammalian cells using a mammalian expression
vector. Examples of mammalian expression vectors include pCDM8
(Seed (1987) Nature 329:840) and pMT2PC (Kaufman et al. (1987) EMBO
J. 6:187-195). When used in mammalian cells, the expression
vector's control functions are often provided by viral regulatory
elements. For example, commonly used promoters are derived from
polyoma, adenovirus 2, cytomegalovirus and simian virus 40. For
other suitable expression systems for both prokaryotic and
eukaryotic cells see chapters 16 and 17 of Sambrook et al.,
supra.
[0263] In another embodiment, the recombinant mammalian expression
vector is capable of directing expression of the nucleic acid
preferentially in a particular cell type (e.g., tissue-specific
regulatory elements are used to express the nucleic acid).
Tissue-specific regulatory elements are known in the art.
Non-limiting examples of suitable tissue-specific promoters include
the albumin promoter (liver-specific; Pinkert et al. (1987) Genes
Dev. 1:268-277), lymphoid-specific promoters (Calame and Eaton
(1988) Adv. Immunol. 43:235-275), in particular promoters of T cell
receptors (Winoto and Baltimore (1989) EMBO J. 8:729-733) and
immunoglobulins (Banerji et al. (1983) Cell 33:729-740; Queen and
Baltimore (1983) Cell 33:741-748), neuron-specific promoters (e.g.,
the neurofilament promoter; Byrne and Ruddle (1989) Proc. Natl.
Acad. Sci. USA 86:5473-5477), pancreas-specific promoters (Edlund
et al. (1985) Science 230:912-916), and mammary gland-specific
promoters (e.g., milk whey promoter; U.S. Pat. No. 4,873,316 and
European Application Publication No. 264,166).
Developmentally-regulated promoters are also encompassed, for
example the murine hox promoters (Kessel and Gruss (1990) Science
249:374-379) and the alpha-fetoprotein promoter (Campes and
Tilghman (1989) Genes Dev. 3:537-546).
[0264] The invention further provides a recombinant expression
vector comprising a DNA molecule of the invention cloned into the
expression vector in an antisense orientation. That is, the DNA
molecule is operably linked with a regulatory sequence in a manner
which allows for expression (by transcription of the DNA molecule)
of an RNA molecule which is antisense, relative to the mRNA
encoding a polypeptide of the invention. Regulatory sequences
operably linked with a nucleic acid cloned in the antisense
orientation can be selected which direct continuous expression of
the antisense RNA molecule in a variety of cell types, for instance
viral promoters and/or enhancers, or regulatory sequences can be
selected which direct constitutive, tissue specific, or cell type
specific expression of antisense RNA. The antisense expression
vector can be in the form of a recombinant plasmid, phagemid, or
attenuated virus in which antisense nucleic acids are produced
under the control of a high efficiency regulatory region, the
activity of which can be determined by the cell type into which the
vector is introduced. For a discussion of the regulation of gene
expression using antisense genes see Weintraub et al.
(Reviews--Trends in Genetics, Vol. 1(1) 1986).
[0265] Another aspect of the invention pertains to host cells into
which a recombinant expression vector of the invention has been
introduced. The terms "host cell" and "recombinant host cell" are
used interchangeably herein. It is understood that such terms refer
not only to the particular subject cell but to the progeny or
potential progeny of such a cell. Because certain modifications can
occur in succeeding generations due to either mutation or
environmental influences, such progeny may not, in fact, be
identical to the parent cell, but are still included within the
scope of the term as used herein.
[0266] A host cell can be any prokaryotic (e.g., E. coli) or
eukaryotic cell (e.g., insect cells, yeast or mammalian cells).
[0267] Vector DNA can be introduced into prokaryotic or eukaryotic
cells via conventional transformation or transfection techniques.
As used herein, the terms "transformation" and "transfection" are
intended to refer to a variety of art-recognized techniques for
introducing foreign nucleic acid into a host cell, including
calcium phosphate or calcium chloride co-precipitation,
DEAE-dextran-mediated transfection, lipofection, and
electroporation. Suitable methods for transforming or transfecting
host cells can be found in Sambrook, et al. (supra), and other
laboratory manuals.
[0268] For stable transfection of mammalian cells, it is known
that, depending upon the expression vector and transfection
technique used, only a small fraction of cells integrate the
foreign DNA into their genome. In order to identify and select
these integrants, a gene that encodes a selectable marker (e.g.,
for resistance to antibiotics) can be introduced into the host
cells along with the gene of interest. Examples of selectable
markers include those which confer resistance to drugs, such as
G418, hygromycin and methotrexate. Cells stably transfected with
the introduced nucleic acid can be identified by drug selection
(e.g., cells that have incorporated the selectable marker gene
survive, while other cells die).
[0269] A host cell of the invention, such as a prokaryotic or
eukaryotic host cell in culture, can be used to produce a
polypeptide of the invention. Accordingly, the invention further
provides methods for producing a polypeptide of the invention using
the host cells of the invention. In one embodiment, the method
comprises culturing the host cell of invention (into which a
recombinant expression vector encoding a polypeptide of the
invention has been introduced) in a suitable medium such that the
polypeptide is produced. In another embodiment, the method further
comprises isolating the polypeptide from the medium or the host
cell.
[0270] The host cells of the invention can be used to produce
non-human transgenic animals. For example, in one embodiment, a
host cell of the invention is a fertilized oocyte or an embryonic
stem cell into which a sequences encoding a polypeptide of the
invention have been introduced. Such host cells can then be used to
create non-human transgenic animals in which exogenous sequences
encoding a polypeptide of the invention have been introduced into
their genome or homologous recombinant animals in which endogenous
encoding a polypeptide of the invention sequences have been
altered. Such animals are useful for studying the function and/or
activity of the polypeptide and for identifying and/or evaluating
modulators of polypeptide activity. As used herein, a "transgenic
animal" is a non-human animal, preferably a mammal, more preferably
a rodent such as a rat or mouse, in which one or more of the cells
of the animal includes a transgene. Other examples of transgenic
animals include non-human primates, sheep, dogs, cows, goats,
chickens, amphibians, etc. A transgene is exogenous DNA which is
integrated into the genome of a cell from which a transgenic animal
develops and which remains in the genome of the mature animal,
thereby directing expression of an encoded gene product in one or
more cell types or tissues of the transgenic animal. As used
herein, an "homologous recombinant animal" is a non-human animal,
preferably a mammal, more preferably a mouse, in which an
endogenous gene has been altered by homologous recombination
between the endogenous gene and an exogenous DNA molecule
introduced into a cell of the animal, e.g., an embryonic cell of
the animal, prior to development of the animal.
[0271] A transgenic animal of the invention can be created by
introducing a nucleic acid encoding a polypeptide of the invention
(or a homologue thereof) into the male pronuclei of a fertilized
oocyte (e.g., by microinjection or retroviral infection) and
allowing the oocyte to develop in a pseudopregnant female foster
animal. Intronic sequences and polyadenylation signals can also be
included in the transgene to increase the efficiency of expression
of the transgene. A tissue-specific regulatory sequence(s) can be
operably linked with the transgene to direct expression of a
polypeptide of the invention to particular cells. Methods for
generating transgenic animals via embryo manipulation and
microinjection, particularly animals such as mice, have become
conventional in the art and are described, for example, in U.S.
Pat. Nos. 4,736,866 and 4,870,009, U.S. Pat. No. 4,873,191 and in
Hogan, Manipulating the Mouse Embryo, (Cold Spring Harbor
Laboratory Press, Cold Spring Harbor, N.Y., 1986). Similar methods
are used for production of other transgenic animals. A transgenic
founder animal can be identified based upon the presence of the
transgene in its genome and/or expression of mRNA encoding the
transgene in tissues or cells of the animals. A transgenic founder
animal can be used to breed additional animals carrying the
transgene. Moreover, transgenic animals harboring the transgene can
further be bred to other transgenic animals harboring other
transgenes.
[0272] To create an homologous recombinant animal, a vector is
prepared which contains at least a portion of a gene encoding a
polypeptide of the invention into which a deletion, addition or
substitution has been introduced to thereby alter, e.g.,
functionally disrupt, the gene. In one embodiment, the vector is
designed such that, upon homologous recombination, the endogenous
gene is functionally disrupted (i.e., no longer encodes a
functional protein; also referred to as a "knock out" vector).
Alternatively, the vector can be designed such that, upon
homologous recombination, the endogenous gene is mutated or
otherwise altered, but still encodes functional protein (e.g., the
upstream regulatory region can be altered to thereby alter the
expression of the endogenous protein). In the homologous
recombination vector, the altered portion of the gene is flanked at
its 5' and 3' ends by additional nucleic acid of the gene to allow
for homologous recombination to occur between the exogenous gene
carried by the vector and an endogenous gene in an embryonic stem
cell. The additional flanking nucleic acid sequences are of
sufficient length for successful homologous recombination with the
endogenous gene. Typically, several kilobases of flanking DNA (both
at the 5' and 3' ends) are included in the vector (see, e.g.,
Thomas and Capecchi (1987) Cell 51:503 for a description of
homologous recombination vectors). The vector is introduced into an
embryonic stem cell line (e.g., by electroporation) and cells in
which the introduced gene has homologously recombined with the
endogenous gene are selected (see, e.g., Li et al. (1992) Cell
69:915). The selected cells are then injected into a blastocyst of
an animal (e.g., a mouse) to form aggregation chimeras (see, e.g.,
Bradley in Teratocarcinomas and Embryonic Stem Cells: A Practical
Approach, Robertson, ed. (IRL, Oxford, 1987) pp. 113-152). A
chimeric embryo can then be implanted into a suitable
pseudopregnant female foster animal and the embryo brought to term.
Progeny harboring the homologously recombined DNA in their germ
cells can be used to breed animals in which all cells of the animal
contain the homologously recombined DNA by germline transmission of
the transgene. Methods for constructing homologous recombination
vectors and homologous recombinant animals are described further in
Bradley (1991) Current Opinion in Bio/Technology 2:823-829 and in
PCT Publication Numbers WO 90/11354, WO 91/01140, WO 92/0968, and
WO 93/04169.
[0273] In another embodiment, transgenic non-human animals can be
produced which contain selected systems which allow for regulated
expression of the transgene. One example of such a system is the
cre/loxP recombinase system of bacteriophage P1. For a description
of the cre/loxP recombinase system, see, e.g., Lakso et al. (1992)
Proc. Natl. Acad. Sci. USA 89:6232-6236. Another example of a
recombinase system is the FLP recombinase system of Saccharomyces
cerevisiae (O'Gorman et al. (1991) Science 251:1351-1355. If a
cre/loxP recombinase system is used to regulate expression of the
transgene, animals containing transgenes encoding both the Cre
recombinase and a selected protein are required. Such animals can
be provided through the construction of "double" transgenic
animals, e.g., by mating two transgenic animals, one containing a
transgene encoding a selected protein and the other containing a
transgene encoding a recombinase.
[0274] Clones of the non-human transgenic animals described herein
can be produced according to the methods described in Wilmut et al.
(1997) Nature 385:810-813 and PCT Publication Numbers WO 97/07668
and WO 97/07669.
IV. Pharmaceutical Compositions
[0275] The nucleic acid molecules, polypeptides, and antibodies
(also referred to herein as "active compounds") of the invention
can be incorporated into pharmaceutical compositions suitable for
administration. Such compositions typically comprise the nucleic
acid molecule, protein, or antibody and a pharmaceutically
acceptable carrier. As used herein the language "pharmaceutically
acceptable carrier" is intended to include any and all solvents,
dispersion media, coatings, anti-bacterial and anti-fungal agents,
isotonic and absorption delaying agents, and the like, compatible
with pharmaceutical administration. The use of such media and
agents for pharmaceutically active substances is well known in the
art. Except insofar as any conventional media or agent is
incompatible with the active compound, use thereof in the
compositions is contemplated. Supplementary active compounds can
also be incorporated into the compositions.
[0276] The invention includes methods for preparing pharmaceutical
compositions for modulating the expression or activity of a
polypeptide or nucleic acid of the invention. Such methods comprise
formulating a pharmaceutically acceptable carrier with an agent
which modulates expression or activity of a polypeptide or nucleic
acid of the invention. Such compositions can further include
additional active agents. Thus, the invention further includes
methods for preparing a pharmaceutical composition by formulating a
pharmaceutically acceptable carrier with an agent which modulates
expression or activity of a polypeptide or nucleic acid of the
invention and one or more additional active compounds.
[0277] The agent which modulates expression or activity can, for
example, be a small molecule. For example, such small molecules
include peptides, peptidomimetics, amino acids, amino acid analogs,
polynucleotides, polynucleotide analogs, nucleotides, nucleotide
analogs, organic or inorganic compounds (i.e., including
heteroorganic and organometallic compounds) having a molecular
weight less than about 10,000 grams per mole, organic or inorganic
compounds having a molecular weight less than about 5,000 grams per
mole, organic or inorganic compounds having a molecular weight less
than about 1,000 grams per mole, organic or inorganic compounds
having a molecular weight less than about 500 grams per mole, and
salts, esters, and other pharmaceutically acceptable forms of such
compounds.
[0278] It is understood that appropriate doses of small molecule
agents and protein or polypeptide agents depends upon a number of
factors within the ken of the ordinarily skilled physician,
veterinarian, or researcher. The dose(s) of these agents will vary,
for example, depending upon the identity, size, and condition of
the subject or sample being treated, further depending upon the
route by which the composition is to be administered, if
applicable, and the effect which the practitioner desires the agent
to have upon the nucleic acid or polypeptide of the invention.
Examples of doses of a small molecule include milligram or
microgram amounts per kilogram of subject or sample weight (e.g.,
about 1 microgram per kilogram to about 500 milligrams per
kilogram, about 100 micrograms per kilogram to about 5 milligrams
per kilogram, or about 1 microgram per kilogram to about 50
micrograms per kilogram). Examples of doses of a protein or
polypeptide include gram, milligram or microgram amounts per
kilogram of subject or sample weight (e.g., about 1 microgram per
kilogram to about 5 grams per kilogram, about 100 micrograms per
kilogram to about 500 milligrams per kilogram, or about 1 milligram
per kilogram to about 50 milligrams per kilogram). For antibodies,
examples of dosages are from about 0.1 milligram per kilogram to
100 milligrams per kilogram of body weight (generally 10 milligrams
per kilogram to 20 milligrams per kilogram). If the antibody is to
act in the brain, a dosage of 50 milligrams per kilogram to 100
milligrams per kilogram is usually appropriate. It is furthermore
understood that appropriate doses of one of these agents depend
upon the potency of the agent with respect to the expression or
activity to be modulated. Such appropriate doses can be determined
using the assays described herein. When one or more of these agents
is to be administered to an animal (e.g., a human) in order to
modulate expression or activity of a polypeptide or nucleic acid of
the invention, a physician, veterinarian, or researcher can, for
example, prescribe a relatively low dose at first, subsequently
increasing the dose until an appropriate response is obtained. In
addition, it is understood that the specific dose level for any
particular animal subject will depend upon a variety of factors
including the activity of the specific agent employed, the age,
body weight, general health, gender, and diet of the subject, the
time of administration, the route of administration, the rate of
excretion, any drug combination, and the degree of expression or
activity to be modulated.
[0279] A pharmaceutical composition of the invention is formulated
to be compatible with its intended route of administration.
Examples of routes of administration include parenteral, e.g.,
intravenous, intradermal, subcutaneous, oral (e.g., inhalation),
transdermal (topical), transmucosal, and rectal administration.
Solutions or suspensions used for parenteral, intradermal, or
subcutaneous application can include the following components: a
sterile diluent such as water for injection, saline solution, fixed
oils, polyethylene glycols, glycerine, propylene glycol or other
synthetic solvents; antibacterial agents such as benzyl alcohol or
methyl parabens; antioxidants such as ascorbic acid or sodium
bisulfite; chelating agents such as ethylenediamine-tetraacetic
acid; buffers such as acetates, citrates or phosphates; and agents
for the adjustment of tonicity such as sodium chloride or dextrose.
pH can be adjusted using acids or bases, such as hydrochloric acid
or sodium hydroxide. The parenteral preparation can be enclosed in
ampules, disposable syringes or multiple dose vials made of glass
or plastic.
[0280] Pharmaceutical compositions suitable for injectable use
include sterile aqueous solutions (where water soluble) or
dispersions and sterile powders for the extemporaneous preparation
of sterile injectable solutions or dispersions. For intravenous
administration, suitable carriers include physiological saline,
bacteriostatic water, Cremophor EL (BASF; Parsippany, N.J.) or
phosphate buffered saline (PBS). The composition should be sterile
and should be fluid to the extent that easy syringability exists.
It should be stable under the conditions of manufacture and storage
and should be preserved against the contaminating action of
microorganisms such as bacteria and fungi. The carrier can be a
solvent or dispersion medium containing, for example, water,
ethanol, polyol (for example, glycerol, propylene glycol, and
liquid polyethylene glycol, and the like), and suitable mixtures
thereof. The proper fluidity can be maintained, for example, by the
use of a coating such as lecithin, by the maintenance of the
required particle size in the case of dispersion and by the use of
surfactants. Prevention of the action of microorganisms can be
achieved by various anti-bacterial and anti-fungal agents, for
example, parabens, chlorobutanol, phenol, ascorbic acid,
thimerosal, and the like. In many cases, it will be preferable to
include isotonic agents, for example, sugars, polyalcohols such as
mannitol, sorbitol, or sodium chloride in the composition.
Prolonged absorption of the injectable compositions can be brought
about by including in the composition an agent which delays
absorption, for example, aluminum monostearate and gelatin.
[0281] Sterile injectable solutions can be prepared by
incorporating the active compound (e.g., a polypeptide or antibody)
in the required amount in an appropriate solvent with one or a
combination of ingredients enumerated above, as required, followed
by filtered sterilization. Generally, dispersions are prepared by
incorporating the active compound into a sterile vehicle which
contains a basic dispersion medium, and then incorporating the
required other ingredients from those enumerated above. In the case
of sterile powders for the preparation of sterile injectable
solutions, examples of methods of preparation are vacuum drying and
freeze-drying which yields a powder of the active ingredient plus
any additional desired ingredient from a previously
sterile-filtered solution thereof.
[0282] Oral compositions generally include an inert diluent or an
edible carrier. They can be enclosed in gelatin capsules or
compressed into tablets. For the purpose of oral therapeutic
administration, the active compound can be incorporated with
excipients and used in the form of tablets, troches, or capsules.
Oral compositions can also be prepared using a fluid carrier for
use as a mouthwash, wherein the compound in the fluid carrier is
applied orally and swished and expectorated or swallowed.
[0283] Pharmaceutically compatible binding agents, adjuvant
materials, or both, can be included as part of the composition. The
tablets, pills, capsules, troches, and the like can contain any of
the following ingredients, or compounds of a similar nature: a
binder such as microcrystalline cellulose, gum tragacanth or
gelatin; an excipient such as starch or lactose, a disintegrating
agent such as alginic acid, Primogel, or corn starch; a lubricant
such as magnesium stearate or Sterotes; a glidant such as colloidal
silicon dioxide; a sweetening agent such as sucrose or saccharin;
or a flavoring agent such as peppermint, methyl salicylate, or
orange flavoring.
[0284] For administration by inhalation, the compounds are
delivered in the form of an aerosol spray from a pressurized
container or dispenser which contains a suitable propellant, e.g.,
a gas such as carbon dioxide, or a nebulizer.
[0285] Systemic administration can also be by transmucosal or
transdermal means. For transmucosal or transdermal administration,
penetrants appropriate to the barrier to be permeated are used in
the formulation. Such penetrants are generally known in the art,
and include, for example, for transmucosal administration,
detergents, bile salts, and fusidic acid derivatives. Transmucosal
administration can be accomplished through the use of nasal sprays
or suppositories. For transdermal administration, the active
compounds are formulated into ointments, salves, gels, or creams as
generally known in the art.
[0286] The compounds can also be prepared in the form of
suppositories (e.g., with conventional suppository bases such as
cocoa butter and other glycerides) or retention enemas for rectal
delivery.
[0287] In one embodiment, the active compounds are prepared with
carriers that will protect the compound against rapid elimination
from the body, such as a controlled release formulation, including
implants and microencapsulated delivery systems. Biodegradable,
biocompatible polymers can be used, such as ethylene vinyl acetate,
polyanhydrides, polyglycolic acid, collagen, polyorthoesters, and
polylactic acid. Methods for preparation of such formulations will
be apparent to those skilled in the art. The materials can also be
obtained commercially from Alza Corporation and Nova
Pharmaceuticals, Inc. Liposomal suspensions (including liposomes
having monoclonal antibodies incorporated therein or thereon) can
also be used as pharmaceutically acceptable carriers. These can be
prepared according to methods known to those skilled in the art,
for example, as described in U.S. Pat. No. 4,522,811.
[0288] It is especially advantageous to formulate oral or
parenteral compositions in dosage unit form for ease of
administration and uniformity of dosage. Dosage unit form as used
herein refers to physically discrete units suited as unitary
dosages for the subject to be treated; each unit containing a
predetermined quantity of active compound calculated to produce the
desired therapeutic effect in association with the required
pharmaceutical carrier. The specification for the dosage unit forms
of the invention are dictated by and directly dependent on the
unique characteristics of the active compound and the particular
therapeutic effect to be achieved, and the limitations inherent in
the art of compounding such an active compound for the treatment of
individuals.
[0289] Generally, partially human antibodies and fully human
antibodies have a longer half-life within the human body than other
antibodies. Accordingly, lower dosages and less frequent
administration is often possible. Modifications such as lipidation
can be used to stabilize antibodies and to enhance uptake and
tissue penetration (e.g., into the brain). A method for lipidation
of antibodies is described by Cruikshank et al. ((1997) J. Acquired
Immune Deficiency Syndromes and Human Retrovirology 14:193).
[0290] The nucleic acid molecules of the invention can be inserted
into vectors and used as gene therapy vectors. Gene therapy vectors
can be delivered to a subject by, for example, intravenous
injection, local administration (U.S. Pat. No. 5,328,470), or by
stereotactic injection (see, e.g., Chen et al. (1994) Proc. Natl.
Acad. Sci. USA 91:3054-3057). The pharmaceutical preparation of the
gene therapy vector can include the gene therapy vector in an
acceptable diluent, or can comprise a slow release matrix in which
the gene delivery vehicle is imbedded. Alternatively, where the
complete gene delivery vector can be produced intact from
recombinant cells, e.g., retroviral vectors, the pharmaceutical
preparation can include one or more cells which produce the gene
delivery system.
[0291] It is recognized that the pharmaceutical compositions and
methods described herein can be used independently or in
combination with one another. That is, subjects can be administered
one or more of the pharmaceutical compositions, e.g.,
pharmaceutical compositions comprising a nucleic acid molecule or
protein of the invention or a modulator thereof, subjected to one
or more of the therapeutic methods described herein, or both, in
temporally overlapping or non-overlapping regimens. When therapies
overlap temporally, the therapies may generally occur in any order
and can be simultaneous (e.g., administered simultaneously together
in a composite composition or simultaneously but as separate
compositions) or interspersed. By way of example, a subject
afflicted with a disorder described herein can be simultaneously or
sequentially administered both a cytotoxic agent which selectively
kills aberrant cells and an antibody (e.g., an antibody of the
invention) which can, in one embodiment, be conjugated or linked
with a therapeutic agent, a cytotoxic agent, an imaging agent, or
the like.
[0292] The pharmaceutical compositions can be included in a
container, pack, or dispenser together with instructions for
administration.
V. Uses and Methods of the Invention
[0293] The nucleic acid molecules, proteins, protein homologs, and
antibodies described herein can be used in one or more of the
following methods: a) screening assays; b) detection assays (e.g.,
chromosomal mapping, tissue typing, forensic biology); c)
predictive medicine (e.g., diagnostic assays, prognostic assays,
monitoring clinical trials, and pharmacogenomics); and d) methods
of treatment (e.g., therapeutic and prophylactic). For example,
polypeptides of the invention can to used for all of the purposes
identified herein in portions of the disclosure relating to
individual types of protein of the invention. The isolated nucleic
acid molecules of the invention can be used to express proteins
(e.g., via a recombinant expression vector in a host cell in gene
therapy applications), to detect mRNA (e.g., in a biological
sample) or a genetic lesion, and to modulate activity of a
polypeptide of the invention. In addition, the polypeptides of the
invention can be used to screen drugs or compounds which modulate
activity or expression of a polypeptide of the invention as well as
to treat disorders characterized by insufficient or excessive
production of a protein of the invention or production of a form of
a protein of the invention which has decreased or aberrant activity
compared to the wild type protein. In addition, the antibodies of
the invention can be used to detect and isolate a protein of the
and modulate activity of a protein of the invention.
[0294] This invention further pertains to novel agents identified
by the above-described screening assays and uses thereof for
treatments as described herein.
A. Screening Assays
[0295] The invention provides a method (also referred to herein as
a "screening assay") for identifying modulators, i.e., candidate or
test compounds or agents (e.g., peptides, peptidomimetics, small
molecules or other drugs) which bind with a polypeptide of the
invention or have a stimulatory or inhibitory effect on, for
example, expression or activity of a polypeptide of the
invention.
[0296] In one embodiment, the invention provides assays for
screening candidate or test compounds which bind with or modulate
the activity of the membrane-bound form of a polypeptide of the
invention or biologically active portion thereof. The test
compounds of the present invention can be obtained using any of the
numerous approaches in combinatorial library methods known in the
art, including: biological libraries; spatially addressable
parallel solid phase or solution phase libraries; synthetic library
methods requiring deconvolution; the "one-bead one-compound"
library method; and synthetic library methods using affinity
chromatography selection. The biological library approach is
limited to peptide libraries, while the other four approaches are
applicable to peptide, non-peptide oligomer, or small molecule
libraries of compounds (Lam (1997) Anticancer Drug Des.
12:145).
[0297] Examples of methods useful for the synthesis of molecular
libraries can be found in the art, for example in: DeWitt et al.
(1993) Proc. Natl. Acad. Sci. USA 90:6909; Erb et al. (1994) Proc.
Natl. Acad. Sci. USA 91:11422; Zuckermann et al. (1994). J. Med.
Chem. 37:2678; Cho et al. (1993) Science 261:1303; Carrell et al.
(1994) Angew. Chem. Int. Ed. Engl. 33:2059; Carell et al. (1994)
Angew. Chem. Int. Ed. Engl. 33:2061; and Gallop et al. (1994) J.
Med. Chem. 37:1233.
[0298] Libraries of compounds can be presented in solution (e.g.,
Houghten (1992) Bio/Techniques 13:412-421), or on beads (Lam (1991)
Nature 354:82-84), chips (Fodor (1993) Nature 364:555-556),
bacteria (U.S. Pat. No. 5,223,409), spores (U.S. Pat. Nos.
5,571,698; 5,403,484; and 5,223,409), plasmids (Cull et al. (1992)
Proc. Natl. Acad. Sci. USA 89:1865-1869) or phage (Scott and Smith
(1990) Science 249:386-390; Devlin (1990) Science 249:404-406;
Cwirla et al. (1990) Proc. Natl. Acad. Sci. USA 87:6378-6382; and
Felici (1991) J. Mol. Biol. 222:301-310).
[0299] In one embodiment, an assay is a cell-based assay in which a
cell which expresses a membrane-bound form of a polypeptide of the
invention, or a biologically active portion thereof, on the cell
surface is contacted with a test compound and the ability of the
test compound to bind with the polypeptide is determined. The cell,
for example, can be a yeast cell or a cell of mammalian origin.
Determining the ability of the test compound to bind with the
polypeptide can be accomplished, for example, by coupling the test
compound with a radioisotope or enzymatic label such that binding
of the test compound to the polypeptide or biologically active
portion thereof can be determined by detecting the labeled compound
in a complex. For example, test compounds can be labeled with
.sup.125I, .sup.35S, .sup.14C, or .sup.3H, either directly or
indirectly, and the radioisotope detected by direct counting of
radio-emission or by scintillation counting. Alternatively, test
compounds can be enzymatically labeled with, for example,
horseradish peroxidase, alkaline phosphatase, or luciferase, and
the enzymatic label detected by determination of conversion of an
appropriate substrate to product. In one embodiment, the assay
comprises contacting a cell which expresses a membrane-bound form
of a polypeptide of the invention, or a biologically active portion
thereof, on the cell surface with a known compound which binds the
polypeptide to form an assay mixture, contacting the assay mixture
with a test compound, and determining the ability of the test
compound to interact with the polypeptide, wherein determining the
ability of the test compound to interact with the polypeptide
comprises determining the ability of the test compound to
preferentially bind with the polypeptide or a biologically active
portion thereof as compared to the known compound.
[0300] In another embodiment, the assay involves assessment of an
activity characteristic of the polypeptide, wherein binding of the
test compound with the polypeptide or a biologically active portion
thereof alters (i.e., increases or decreases) the activity of the
polypeptide.
[0301] In another embodiment, an assay is a cell-based assay
comprising contacting a cell expressing a membrane-bound form of a
polypeptide of the invention, or a biologically active portion
thereof, on the cell surface with a test compound and determining
the ability of the test compound to modulate (e.g., stimulate or
inhibit) the activity of the polypeptide or biologically active
portion thereof. Determining the ability of the test compound to
modulate the activity of the polypeptide or a biologically active
portion thereof can be accomplished, for example, by determining
the ability of the polypeptide to bind with or interact with a
target molecule or to transport molecules across the cytoplasmic
membrane.
[0302] Determining the ability of a polypeptide of the invention to
bind with or interact with a target molecule can be accomplished by
one of the methods described above for determining direct binding.
As used herein, a "target molecule" is a molecule with which a
selected polypeptide (e.g., a polypeptide of the invention binds or
interacts with in nature, for example, a molecule on the surface of
a cell which expresses the selected protein, a molecule on the
surface of a second cell, a molecule in the extracellular milieu, a
molecule associated with the internal surface of a cell membrane or
a cytoplasmic molecule. A target molecule can be a polypeptide of
the invention or some other polypeptide or protein. For example, a
target molecule can be a component of a signal transduction pathway
which facilitates transduction of an extracellular signal (e.g., a
signal generated by binding of a compound to a polypeptide of the
invention) through the cell membrane and into the cell or a second
intercellular protein which has catalytic activity or a protein
which facilitates association of downstream signaling molecules
with a polypeptide of the invention. Determining the ability of a
polypeptide of the invention to bind with or interact with a target
molecule can be accomplished by determining the activity of the
target molecule. For example, the activity of the target molecule
can be determined by detecting induction of a cellular second
messenger of the target (e.g., an mRNA, intracellular Ca.sup.2+,
diacylglycerol, IP3, and the like), detecting catalytic/enzymatic
activity of the target on an appropriate substrate, detecting
induction of a reporter gene (e.g., a regulatory element that is
responsive to a polypeptide of the invention operably linked with a
nucleic acid encoding a detectable marker, e.g., luciferase), or
detecting a cellular response, for example, cellular
differentiation, or cell proliferation.
[0303] In yet another embodiment, an assay of the present invention
is a cell-free assay comprising contacting a polypeptide of the
invention or biologically active portion thereof with a test
compound and determining the ability of the test compound to bind
with the polypeptide or biologically active portion thereof.
Binding of the test compound with the polypeptide can be determined
either directly or indirectly as described above. In one
embodiment, the assay includes contacting the polypeptide of the
invention or biologically active portion thereof with a known
compound which binds the polypeptide to form an assay mixture,
contacting the assay mixture with a test compound, and determining
the ability of the test compound to interact with the polypeptide,
wherein determining the ability of the test compound to interact
with the polypeptide comprises determining the ability of the test
compound to preferentially bind with the polypeptide or
biologically active portion thereof as compared to the known
compound.
[0304] In another embodiment, an assay is a cell-free assay
comprising contacting a polypeptide of the invention or
biologically active portion thereof with a test compound and
determining the ability of the test compound to modulate (e.g.,
stimulate or inhibit) the activity of the polypeptide or
biologically active portion thereof. Determining the ability of the
test compound to modulate activity of the polypeptide can be
accomplished, for example, by determining the ability of the
polypeptide to bind with a target molecule by one of the methods
described above for determining direct binding. In an alternative
embodiment, determining the ability of the test compound to
modulate the activity of the polypeptide can be accomplished by
determining the ability of the polypeptide of the invention to
further modulate the target molecule. For example, the catalytic
activity, the enzymatic activity, or both, of the target molecule
on an appropriate substrate can be determined as previously
described.
[0305] In yet another embodiment, the cell-free assay comprises
contacting a polypeptide of the invention or biologically active
portion thereof with a known compound which binds the polypeptide
to form an assay mixture, contacting the assay mixture with a test
compound, and determining the ability of the test compound to
interact with the polypeptide. Ability of the test compound to
interact with the polypeptide can be determined by assessing the
ability of the polypeptide to preferentially bind with or modulate
the activity of a target molecule, or by any other method.
[0306] The cell-free assays of the present invention are amenable
to use of either soluble or membrane-bound forms (where applicable)
of a polypeptide of the invention. In the case of cell-free assays
comprising a membrane-bound form of the polypeptide, it can be
desirable to use a solubilizing agent in order to maintain the
membrane-bound form of the polypeptide in solution. Examples of
such solubilizing agents include non-ionic detergents such as
n-octylglucoside, n-dodecylglucoside, n-octylmaltoside,
octanoyl-N-methylglucamide, decanoyl-N-methylglucamide, Triton
X-100, Triton X-114, Thesit, isotridecypoly(ethylene glycol
ether)n, 3-{(3-cholamidopropyl) dimethylamminio}-1-propane
sulfonate (CHAPS), 3-{(3-cholamidopropyl)
dimethylamminio}-2-hydroxy-1-propane sulfonate (CHAPSO), or
N-dodecyl-N,N-dimethyl-3-ammonio-1-propane sulfonate.
[0307] In one or more embodiments of the above assay methods of the
present invention, it can be desirable to immobilize either the
polypeptide of the invention or its target molecule in order to
facilitate separation of complexed and non-complexed forms of one
or both of the molecules, as well as to accommodate automation of
the assay. Binding of a test compound with the polypeptide, or
interaction of the polypeptide with a target molecule in the
presence and absence of a candidate compound, can be accomplished
in any vessel suitable for containing the reactants. Examples of
such vessels include microtiter plates, test tubes, and
micro-centrifuge tubes. In one embodiment, a fusion protein can be
provided which adds a domain that allows one or both of the
proteins to be bound to a matrix. For example,
glutathione-S-transferase fusion proteins or
glutathione-S-transferase fusion proteins can be adsorbed onto
glutathione Sepharose.TM. beads (Sigma Chemical; St. Louis, Mo.) or
glutathione-derivatized microtiter plates, which are combined with
the test compound and either the non-adsorbed target protein or a
polypeptide of the invention. The combination is incubated under
conditions conducive to complex formation (e.g., at physiological
conditions for salt and pH). Following incubation, the beads or
microtiter plate wells are washed to remove unbound components, and
complex formation is measured directly or indirectly, for example,
as described above. Alternatively, the complexes can be dissociated
from the matrix, and the level of binding or activity of the
polypeptide of the invention can be determined using standard
techniques, such as those described herein.
[0308] Other techniques for immobilizing proteins on matrices can
also be used in the screening assays of the invention. For example,
either the polypeptide of the invention or a target molecule
thereof (e.g., a protein which binds therewith or a substrate or an
analog of a substrate of the protein of the invention) can be
immobilized using conjugation of biotin and streptavidin.
Biotinylated polypeptide of the invention or target molecules can
be prepared using biotin-NHS (biotin-N-hydroxy-succinimide) using
techniques well known in the art (e.g., using a commercially
available kit such as the biotinylation kit manufactured by Pierce
Chemical Co.; Rockford, Ill.), and immobilized in the wells of
streptavidin-coated 96-well plates (Pierce Chemical).
Alternatively, antibodies which are reactive with the polypeptide
of the invention or target molecules but which do not interfere
with binding of the polypeptide of the invention with its target
molecule can be derivatized to the wells of the plate, and unbound
target or polypeptide of the invention can be trapped in the wells
by antibody conjugation. Methods for detecting such complexes, in
addition to those described above for the GST-immobilized
complexes, include immunodetection of complexes using antibodies
reactive with the polypeptide of the invention or target molecule,
as well as enzyme-linked assays which rely on detecting an
enzymatic activity associated with the polypeptide of the invention
or target molecule.
[0309] In another embodiment, modulators of expression of a
polypeptide of the invention are identified in a method in which a
cell is contacted with a candidate compound and expression of the
selected mRNA or protein (i.e., mRNA or protein corresponding to a
polypeptide or nucleic acid of the invention) in the cell is
determined. The level of expression of the selected mRNA or protein
in the presence of the candidate compound is compared with the
level of expression of the selected mRNA or protein in the absence
of the candidate compound. The candidate compound can then be
identified as a modulator of expression of the polypeptide of the
invention based on this comparison. For example, if expression of
the selected mRNA or protein is greater (i.e., statistically
significantly greater) in the presence of the candidate compound
than in its absence, then the candidate compound is identified as a
stimulator of expression of the selected mRNA or protein.
Alternatively, if expression of the selected mRNA or protein is
less (i.e., statistically significantly less) in the presence of
the candidate compound than in its absence, then the candidate
compound is identified as an inhibitor of expression of the
selected mRNA or protein. The level of the selected mRNA or protein
expression in the cells can be determined by methods described
herein.
[0310] In yet another aspect of the invention, a polypeptide of the
invention can be used as a "bait protein" in a two-hybrid assay or
three hybrid assay (see, e.g., U.S. Pat. No. 5,283,317; Zervos et
al. (1993) Cell 72:223-232; Madura et al. (1993) J. Biol. Chem.
268:12046-12054; Bartel et al. (1993) Bio/Techniques 14:920-924;
Iwabuchi et al. (1993) Oncogene 8:1693-1696; and PCT Publication
No. WO 94/10300), to identify other proteins which bind with or
interact with the polypeptide of the invention and modulate
activity of the polypeptide of the invention. Such binding proteins
are also likely to be involved in the propagation of signals by the
polypeptide of the inventions as, for example, upstream or
downstream elements of a signaling pathway involving the
polypeptide of the invention.
[0311] This invention further pertains to novel agents identified
by the above-described screening assays and uses thereof for
treatments as described herein.
B. Detection Assays
[0312] Portions or fragments of the cDNA sequences identified
herein (and the corresponding complete gene sequences) can be used
in numerous ways as polynucleotide reagents. For example, these
sequences can be used to: (i) map their respective genes on a
chromosome and, thus, locate gene regions associated with genetic
disease; (ii) identify an individual from a minute biological
sample (tissue typing); and (iii) aid in forensic identification of
a biological sample. These applications are described in the
subsections below.
1. Chromosome Mapping
[0313] Once the sequence (or a portion of the sequence) of a gene
has been isolated, this sequence can be used to map the location of
the gene on a chromosome. Accordingly, nucleic acid molecules
described herein or fragments thereof, can be used to map the
location of the corresponding genes on a chromosome. Mapping of
sequences to chromosomes is an important first step in correlating
these sequences with genes associated with occurrence of disease.
For example, the TANGO 273 gene exhibits significant homology with
a portion of chromosome 7 between chromosomal markers D7S2467 and
D7S2552.
[0314] Briefly, genes can be mapped to chromosomes by preparing PCR
primers (preferably 15-25 nucleotide residues in length) from the
sequence of a gene of the invention. Computer analysis of the
sequence of a gene of the invention can be used to rapidly select
primers that do not span more than one exon in the genomic DNA,
which would complicate the amplification process. These primers can
be used for PCR screening of somatic cell hybrids containing
individual human chromosomes. Only those hybrids containing the
human gene corresponding to the gene sequences will yield an
amplified fragment. For a review of this technique, see D'Eustachio
et al. ((1983) Science 220:919-924).
[0315] PCR mapping of somatic cell hybrids is a rapid procedure for
assigning a particular sequence to a particular chromosome. Three
or more sequences can be assigned per day using a single thermal
cycler. Using one or more nucleic acid sequences of the invention
to design oligonucleotide primers, sub-localization can be achieved
using panels of fragments prepared from specific chromosomes. Other
mapping strategies which can similarly be used to map a gene to its
chromosomal location include in situ hybridization (described in
Fan et al. (1990) Proc. Natl. Acad. Sci. USA 87:6223-27),
pre-screening with labeled flow-sorted chromosomes, and
pre-selection by hybridization with chromosome specific cDNA
libraries. Fluorescence in situ hybridization (FISH) of a DNA
sequence using a metaphase chromosomal spread can be used to
provide a precise chromosomal location in one step. For a review of
this technique, see Verma et al. (Human Chromosomes: A Manual of
Basic Techniques (Pergamon Press, New York, 1988)).
[0316] Reagents for chromosome mapping can be used individually to
mark a single chromosome or a single site on a chromosome.
Alternatively, panels of reagents can be used for marking multiple
sites, multiple chromosomes, or both. Reagents corresponding to
non-coding regions of the genes actually are preferred for mapping
purposes. Coding sequences are more likely to be conserved within
gene families, thus increasing the chance of cross-hybridization
during chromosomal mapping.
[0317] Once a sequence has been mapped to a precise chromosomal
location, the physical position of the sequence on the chromosome
can be correlated with genetic map data. (Such data are found, for
example, in V. McKusick, Mendelian Inheritance in Man, available
on-line through Johns Hopkins University Welch Medical Library).
The relationship between genes and disease, mapped to the same
chromosomal region, can then be identified by linkage analysis
(co-inheritance of physically adjacent genes), described in, e.g.,
Egeland et al. (1987) Nature 325:783-787.
[0318] Moreover, differences in the DNA sequences between
individuals affected and non-affected with a disease associated
with a gene of the invention can be determined. If a mutation is
observed in some or all of the affected individuals, but not in any
(or in very few) non-affected individuals, then the mutation is
likely to be the causative agent of the particular disease.
Comparison of affected and non-affected individuals generally
involves first looking for structural alterations in the
chromosomes such as deletions or translocations that are visible
from chromosome spreads or detectable using PCR based on that DNA
sequence. Ultimately, complete sequencing of genes from several
individuals can be performed to confirm the presence of a mutation
and to distinguish mutations from polymorphisms.
2. Tissue Typing
[0319] The nucleic acid sequences of the present invention can also
be used to identify individuals from minute biological samples. The
United States military, for example, is considering the use of
restriction fragment length polymorphism (RFLP) for identification
of its personnel. In this technique, an individual's genomic DNA is
digested with one or more restriction enzymes, and probed on a
Southern blot to yield unique bands for identification. This method
does not suffer from the current limitations of physical
identification devices such as general issue "dog tags," which can
be lost, switched, or stolen, making positive identification
difficult. The sequences of the present invention are useful as
additional DNA markers for RFLP (described in U.S. Pat. No.
5,272,057).
[0320] Furthermore, the sequences of the present invention can be
used to provide an alternative technique which determines the
actual base-by-base DNA sequence of selected portions of an
individual's genome. The nucleic acid sequences described herein
can be used to prepare two PCR primers from the 5' and 3' ends of
the sequences. These primers can then be used to amplify an
individual's DNA and to subsequently sequence it.
[0321] Panels of corresponding DNA sequences from individuals,
prepared in this manner, can provide unique individual
identifications, because (with the exception of identical twins)
every individual has a unique set of such DNA sequences owing, at
least in part, to allelic differences. Sequences of the present
invention can be used to obtain such identification sequences from
individuals and from tissue. The nucleic acid sequences of the
invention uniquely represent portions of the human genome. Allelic
variation occurs to some degree in the coding regions of these
sequences, and to a greater degree in the non-coding regions. It is
estimated that allelic variation between individual humans occurs
with a frequency of about once per 500 nucleotide residues. Each of
the sequences described herein can, to some degree, be used as a
standard against which DNA from an individual can be compared for
identification purposes. Because greater numbers of polymorphisms
occur in the non-coding regions, fewer non-coding sequences are
necessary to differentiate individuals. The non-coding sequences of
any of SEQ ID NOs: 1, 11, 21, 31, 41, 51, 61, 71, and 81 can
comfortably provide positive individual identification with a panel
of perhaps 10 to 1,000 primers which each yield a non-coding
amplified sequence of 100 bases. If predicted coding sequences,
such as those in any of SEQ ID NOs: 2, 12, 22, 32, 42, 52, 62, 72,
and 82 are used, a more appropriate number of primers for positive
individual identification would be 500-2,000.
[0322] If a panel of reagents from the nucleic acid sequences
described herein is used to generate a unique identification
database for an individual, those same reagents can later be used
to identify nucleic acids, cells, or tissue from that individual.
Using the unique identification database, positive identification
of the individual, living or dead, can be made from extremely small
samples.
3. Use of Partial Gene Sequences in Forensic Biology
[0323] DNA-based identification techniques can be used in forensic
biology. Forensic biology is a scientific field employing genetic
typing of biological evidence found at a crime scene as a means for
positively identifying, for example, a perpetrator of a crime. To
make such an identification, PCR technology can be used to amplify
DNA sequences taken from very small biological samples such as
tissues (e.g., hair or skin) or body fluids (e.g., blood, saliva,
or semen) found at a crime scene. The amplified sequence can be
compared with a standard, thereby allowing identification of the
origin of the biological sample.
[0324] The sequences of the present invention can be used to
provide polynucleotide reagents (e.g., PCR primers) targeted to
specific loci in the human genome, which can enhance the
reliability of DNA-based forensic identifications by, for example,
providing another "identification marker" (i.e., another DNA
sequence that is unique to a particular individual). As mentioned
above, actual nucleotide sequence information can be used for
identification as an accurate alternative to patterns formed by
restriction enzyme-generated fragments. Sequences of non-coding
regions are particularly appropriate for this use, because greater
numbers of polymorphisms occur in non-coding regions, making it
easier to differentiate individuals using this technique. Examples
of polynucleotide reagents include the nucleic acid sequences of
the invention or portions thereof, e.g., fragments derived from
non-coding regions having a length of at least 20 or 30 nucleotide
residues.
[0325] The nucleic acid sequences described herein can further be
used to provide polynucleotide reagents, e.g., labeled or labelable
probes which can be used in, for example, an in situ hybridization
technique, to identify a specific tissue, e.g., brain tissue. This
can be very useful in cases where a forensic pathologist is
presented with a tissue of unknown origin. Panels of such probes
can be used to identify tissue by species and/or by organ type.
C. Predictive Medicine
[0326] The present invention also pertains to the field of
predictive medicine in which diagnostic assays, prognostic assays,
and monitoring clinical trials are used for prognostic (predictive)
purposes to thereby treat an individual prophylactically.
Accordingly, one aspect of the present invention relates to
diagnostic assays for determining expression of a gene encoding a
polypeptide of the invention as well as activity of a polypeptide
of the invention, in the context of a biological sample (e.g.,
blood, serum, cells, tissue) to thereby determine whether an
individual is afflicted with a disease or disorder, or is at risk
of developing a disorder, associated with aberrant or unwanted
expression of a gene encoding a polypeptide of the invention or
aberrant or unwanted activity of a polypeptide of the invention.
The invention also provides for prognostic (or predictive) assays
for determining whether an individual is at risk of developing a
disorder associated with a protein of the invention, with
expression of a nucleic acid encoding a polypeptide of the
invention, or with activity of a polypeptide of the invention. For
example, mutations in a gene encoding a polypeptide of the
invention can be assayed in a biological sample. Such assays can be
used for prognostic or predictive purpose to thereby
prophylactically treat an individual prior to the onset of a
disorder characterized by or associated with a polypeptide of the
invention, expression of a nucleic acid encoding it, or its
activity.
[0327] As an alternative to making determinations based on the
absolute expression level of selected genes, determinations may be
based on the normalized expression levels of these genes.
Expression levels are normalized by correcting the absolute
expression level of a gene encoding a polypeptide of the invention
by comparing its expression to the expression of a different gene,
e.g., a housekeeping gene that is constitutively expressed.
Suitable genes for normalization include housekeeping genes such as
the actin gene. This normalization allows the comparison of the
expression level in one sample (e.g., a patient sample), to another
sample, or between samples from different sources.
[0328] Alternatively, the expression level can be provided as a
relative expression level. To determine a relative expression level
of a gene, the level of expression of the gene is determined for 10
or more samples of different endothelial (e.g., intestinal
endothelium, airway endothelium, or other mucosal epithelium) cell
isolates, preferably 50 or more samples, prior to the determination
of the expression level for the sample in question. The mean
expression level of each of the genes assayed in the larger number
of samples is determined and this is used as a baseline expression
level for the gene(s) in question. The expression level of the gene
determined for the test sample (absolute level of expression) is
then divided by the mean expression value obtained for that gene.
This provides a relative expression level and aids in identifying
extreme cases of disorders associated with aberrant expression of a
gene encoding a polypeptide of the invention protein or with
aberrant expression of a ligand thereof.
[0329] Preferably, the samples used in the baseline determination
will be from either or both of cells which aberrantly express a
gene encoding a polypeptide of the invention or a ligand thereof
(i.e. `diseased cells`) and cells which express a gene encoding a
polypeptide of the invention at a normal level or a ligand thereof
(i.e. `normal` cells). The choice of the cell source is dependent
on the use of the relative expression level. Using expression found
in normal tissues as a mean expression score aids in validating
whether aberrance in expression of a gene encoding a polypeptide of
the invention occurs specifically in diseased cells. Such a use is
particularly important in identifying whether a gene encoding a
polypeptide of the invention can serve as a target gene. In
addition, as more data is accumulated, the mean expression value
can be revised, providing improved relative expression values based
on accumulated data. Expression data from endothelial cells (e.g.,
mucosal endothelial cells) provides a means for grading the
severity of the disorder.
[0330] Another aspect of the invention pertains to monitoring the
influence of agents (e.g., drugs, antibodies, antisense
oligonucleotides, or other compounds) on the expression or activity
of a polypeptide of the invention in clinical trials.
[0331] These and other agents are described in further detail in
the following sections.
1. Diagnostic Assays
[0332] An example of a method for detecting the presence or absence
of a polypeptide or nucleic acid of the invention in a biological
sample involves obtaining a biological sample from a test subject
and contacting the biological sample with a compound or an agent
capable of detecting a polypeptide or nucleic acid (e.g., mRNA,
genomic DNA) of the invention. An example of an agent for detecting
mRNA or genomic DNA encoding a polypeptide of the invention is a
labeled nucleic acid probe capable of hybridizing with mRNA or
genomic DNA encoding a polypeptide of the invention. The nucleic
acid probe can be, for example, a full-length cDNA, such as the
nucleic acid of one of SEQ ID NOs: 1, 11, 21, 31, 41, 51, 61, 71,
and 81, or a portion thereof, such as an oligonucleotide of at
least 15, 30, 50, 100, 250 or 500 nucleotides in length and
sufficient to specifically hybridize under stringent conditions
with a mRNA or genomic DNA encoding a polypeptide of the invention.
Other suitable probes for use in the diagnostic assays of the
invention are described herein.
[0333] An example of an agent for detecting a polypeptide of the
invention is an antibody capable of binding with a polypeptide of
the invention, such as an antibody having a detectable label.
Antibodies can be polyclonal or, preferably, monoclonal. An intact
antibody, or a fragment thereof (e.g., Fab or F(ab')2) can be used.
The term "labeled," with regard to the probe or antibody, includes
direct labeling of the probe or antibody by coupling (i.e.,
physically linking) a detectable substance to the probe or
antibody, as well as indirect labeling of the probe or antibody by
coupling it with another reagent that is directly labeled. Examples
of indirect labeling include detection of a primary antibody using
a fluorescently labeled secondary antibody and end-labeling of a
DNA probe with biotin such that it can be detected with
fluorescently labeled streptavidin. The term "biological sample" is
intended to include tissues, cells, and biological fluids isolated
from a subject, as well as tissues, cells, and fluids present
within a subject. That is, the detection method of the invention
can be used to detect mRNA, protein, or genomic DNA in a biological
sample in vitro as well as in vivo. For example, in vitro
techniques for detection of mRNA include Northern hybridization
methods and in situ hybridization methods. In vitro techniques for
detection of a polypeptide of the invention include enzyme linked
immunosorbent assays (ELISAs), Western blots, immunoprecipitation,
and immunofluorescence. In vitro techniques for detection of
genomic DNA include Southern hybridizations. Furthermore, in vivo
techniques for detection of a polypeptide of the invention include
introducing into a subject a labeled antibody directed against the
polypeptide. For example, the antibody can be labeled with a
radioactive marker, the presence and location of which in a subject
can be detected using standard imaging techniques.
[0334] In one embodiment, the biological sample contains protein
molecules obtained from the test subject. Alternatively, the
biological sample can contain mRNA molecules obtained from the test
subject or genomic DNA molecules obtained from the test subject. An
example of a biological sample is a peripheral blood
leukocyte-containing sample obtained by conventional means from a
subject (e.g., isolated peripheral blood leukocytes).
[0335] In another embodiment, the methods further involve obtaining
a control biological sample from a control (i.e., non-afflicted)
subject, contacting the control sample with a compound or agent
capable of detecting a polypeptide of the invention or mRNA or
genomic DNA encoding a polypeptide of the invention. The presence
or amount of the polypeptide, mRNA, or genomic DNA encoding the
polypeptide in the control and test samples can be compared to
assess the degree, if any, to which the presence or amount in the
test sample differs from that in the control sample.
[0336] The invention also encompasses kits for detecting the
presence of a polypeptide or nucleic acid of the invention in a
biological sample obtained from a subject. Such kits can be used to
determine if a subject is suffering from or is at increased risk of
developing a disorder associated with aberrant expression of a
polypeptide of the invention (e.g., one of the disorders described
in the section of this disclosure wherein the individual
polypeptide of the invention is discussed). For example, the kit
can comprise a labeled compound or agent capable of detecting the
polypeptide or mRNA encoding the polypeptide in a biological
sample. The kit can also, or alternatively, contain means for
determining the amount of the polypeptide or mRNA in the sample
(e.g., an antibody which specifically binds with the polypeptide or
an oligonucleotide probe which binds with a nucleic acid encoding
the polypeptide). Kits can include instructions for assessing
whether the tested subject is suffering from or is at risk of
developing a disorder associated with aberrant expression of the
polypeptide if the amount of the polypeptide or mRNA encoding the
polypeptide is above or below a normal level.
[0337] For antibody-based kits, the kit can comprise, for example:
(1) a first antibody (e.g., attached to a solid support) which
specifically binds with a polypeptide of the invention; and,
optionally, (2) a second, different antibody which specifically
binds with either the polypeptide or the first antibody and is
conjugated with a detectable agent.
[0338] For oligonucleotide-based kits, the kit can comprise, for
example: (1) an oligonucleotide (e.g., a detectably labeled
oligonucleotide) which hybridizes with a nucleic acid encoding a
polypeptide of the invention or (2) a pair of primers useful for
amplifying a nucleic acid encoding a polypeptide of the invention.
The kit can comprise, for example, a buffering agent, a
preservative, or a protein stabilizing agent. The kit can also
comprise components necessary for detecting the detectable agent
(e.g., an enzyme or a substrate). The kit can contain a control
sample or a series of control samples which can be assayed and
compared with the test sample assay results. Each component of the
kit can be enclosed within an individual container and all of the
various containers can furthermore be within a single package,
optionally with instructions for assessing whether the tested
subject is suffering from or is at risk of developing a disorder
associated with aberrant expression of the polypeptide.
2. Prognostic Assays
[0339] The methods described herein can furthermore be used as
diagnostic or prognostic assays to identify subjects having or at
risk of developing a disease or disorder associated with aberrant
expression or activity of a polypeptide of the invention (e.g., one
of the disorders described in the section of this disclosure
wherein the individual polypeptide of the invention is discussed).
Thus, the present invention provides a method in which a test
sample is obtained from a subject and a polypeptide or nucleic acid
(e.g., mRNA, genomic DNA) of the invention is detected, wherein the
presence, level, or activity of the polypeptide or nucleic acid in
the sample is associated with an enhanced or diminished risk of
developing a disease or disorder associated with aberrant
expression or activity of the polypeptide.
[0340] Furthermore, the prognostic assays described herein can be
used to determine whether an agent (e.g., an agonist, antagonist,
peptidomimetic, protein, peptide, nucleic acid, small molecule, or
other drug candidate) can be administered to a subject in order to
treat a disease or disorder associated with aberrant expression or
activity of a polypeptide of the invention. For example, such
methods can be used to determine whether a subject can be
effectively treated using a specific agent or class of agents
(e.g., agents of a type which decrease activity of the
polypeptide). Thus, the present invention provides methods for
determining whether an agent can be administered to a subject in
order to effectively treat a disorder associated with aberrant
expression or activity of a polypeptide of the invention. When
efficacious agents are known or found, such assays can also be used
to estimate tan efficacious dose of the agent.
[0341] The methods of the invention can be used to detect genetic
lesions or mutations in a gene of the invention in order to assess
if a subject having the lesioned or mutated gene is at risk for a
disorder characterized aberrant expression or activity of a
polypeptide of the invention. In certain embodiments, the methods
include detecting, in a sample of cells obtained from the subject,
the presence or absence of a genetic lesion or mutation
characterized by at least one of an alteration affecting the
integrity of a gene encoding the polypeptide of the invention, or
the mis-expression of the gene encoding the polypeptide of the
invention. For example, such genetic lesions or mutations can be
detected by ascertaining the existence of at least one of: 1) a
deletion of one or more nucleotides from the gene; 2) an addition
of one or more nucleotides to the gene; 3) a substitution of one or
more nucleotides of the gene; 4) a chromosomal rearrangement of the
gene; 5) an alteration in the level of a messenger RNA transcript
of the gene; 6) an aberrant modification of the gene, such as of
the methylation pattern of the genomic DNA; 7) a non-wild type
splicing pattern of a messenger RNA transcript of the gene; 8) a
non-wild type level of the protein encoded by the gene; 9) an
allelic loss of the gene; and 10) an inappropriate
post-translational modification of the protein encoded by the gene.
As described herein, there are a large number of assay techniques
known in the art which can be used for detecting such lesions and
mutations in a gene.
[0342] In certain embodiments, detection of the lesion involves the
use of an oligonucleotide primer in a polymerase chain reaction
(PCR; see, e.g., U.S. Pat. Nos. 4,683,195 and 4,683,202), such as
anchor PCR or RACE PCR, or, alternatively, in a ligation chain
reaction (LCR; see, e.g., Landegran et al. (1988) Science
241:1077-1080; and Nakazawa et al. (1994) Proc. Natl. Acad. Sci.
USA 91:360-364), the latter of which can be particularly useful for
detecting point mutations in a gene (see, e.g., Abravaya et al.
(1995) Nucleic Acids Res. 23:675-682). This method can include the
steps of collecting a sample of cells from a patient, isolating
nucleic acid (e.g., genomic, mRNA, or both) from the cells of the
sample, contacting the nucleic acid sample with one or more primers
which specifically hybridize with the selected gene under
conditions such that hybridization and amplification of the gene
(if present) occurs, and detecting the presence or absence of an
amplification product. The method can also include detecting the
size of the amplification product and comparing the length to the
length of a corresponding product obtained in the same manner from
a control sample. PCR, LCR, or both can be used as a preliminary
amplification step in conjunction with any of the techniques used
for detecting mutations described herein.
[0343] Alternative amplification methods include: self-sustained
sequence replication (Guatelli et al. (1990) Proc. Natl. Acad. Sci.
USA 87:1874-1878), transcriptional amplification system (Kwoh, et
al. (1989) Proc. Natl. Acad. Sci. USA 86:1173-1177), Q-Beta
Replicase (Lizardi et al. (1988) Bio/Technology 6:1197), or any
other nucleic acid amplification method, followed by the detection
of the amplified molecules using any of a variety of techniques
well known to those of skill in the art. These detection schemes
are especially useful for detection of nucleic acid molecules if
such molecules are present in very low numbers.
[0344] In an alternative embodiment, mutations in a selected gene
can be identified in a sample by detecting alterations in
restriction enzyme cleavage patterns. For example, sample and
control DNA is isolated, (optionally) amplified, digested with one
or more restriction endonucleases, and fragment length sizes are
determined by gel electrophoresis and compared. Differences in
fragment length sizes between sample and control DNA indicates
occurrence of mutations or other sequence differences in the sample
DNA. Moreover, sequence specific ribozymes (see, e.g., U.S. Pat.
No. 5,498,531) can be used to score for the presence of specific
mutations by development or loss of a ribozyme cleavage site.
[0345] In other embodiments, genetic mutations are identified by
hybridizing a sample and control nucleic acids, e.g., DNA or RNA,
with high density arrays containing hundreds or thousands of
oligonucleotides probes (Cronin et al. (1996) Human Mutation
7:244-255; Kozal et al. (1996) Nature Medicine 2:753-759). For
example, genetic mutations can be identified using two-dimensional
arrays of light-generated DNA probes fixed to a surface, as
described in Cronin et al., supra. Briefly, a first hybridization
array of probes can be used to scan through long stretches of DNA
in a sample and control to identify base changes between the
sequences by making linear arrays of sequential overlapping probes.
This step allows the identification of point mutations. This step
is followed by hybridization of the nucleic acid sample with a
second hybridization array in order to characterize specific
mutations using smaller, specialized probe arrays complementary to
many or all potential variants or mutations. Each mutation array is
composed of parallel probe sets, one complementary to the wild-type
gene and the other complementary to the mutant gene.
[0346] In yet another embodiment, any of a variety of sequencing
methods known in the art can be used to directly sequence the
selected gene and detect mutations by comparing the sequence of the
sample nucleic acids with the corresponding wild-type (control)
sequence. Examples of sequencing reactions include those based on
techniques developed by Maxim and Gilbert ((1977) Proc. Natl. Acad.
Sci. USA 74:560) or Sanger ((1977) Proc. Natl. Acad. Sci. USA
74:5463). It is also contemplated that any of a variety of
automated sequencing procedures can be used when performing the
diagnostic assays ((1995) Bio/Techniques 19:448), including
sequencing by mass spectrometry (see, e.g., PCT Publication No. WO
94/16101; Cohen et al. (1996) Adv. Chromatogr. 36:127-162; and
Griffin et al. (1993) Appl. Biochem. Biotechnol. 38:147-159).
[0347] Other methods for detecting mutations in a selected gene
include methods in which protection from cleavage agents is used to
detect mismatched bases in RNA/RNA or RNA/DNA heteroduplexes (Myers
et al. (1985) Science 230:1242). In general, the technique of
mismatch cleavage entails providing heteroduplexes formed by
hybridizing (labeled) RNA or DNA containing the wild-type sequence
with potentially mutant RNA or DNA obtained from a tissue sample.
The double-stranded duplexes are treated with an agent that cleaves
single-stranded regions of the duplex such as those which exist due
to base pair mismatches between the control and sample strands.
RNA/DNA duplexes can be treated with RNase to digest mismatched
regions, and DNA/DNA hybrids can be treated with S1 nuclease to
digest mismatched regions.
[0348] In other embodiments, DNA/DNA or RNA/DNA duplexes can be
treated with hydroxylamine or osmium tetroxide and with piperidine
in order to digest mismatched regions. After digestion of the
mismatched regions, the resulting material is separated by size on
denaturing polyacrylamide gels to determine the site of the mutated
or mismatched region. See, e.g., Cotton et al. (1988) Proc. Natl.
Acad. Sci. USA 85:4397; Saleeba et al. (1992) Methods Enzymol.
217:286-295. In one embodiment, the control DNA or RNA is labeled
for detection.
[0349] In still another embodiment, the mismatch cleavage reaction
employs one or more proteins that recognize mismatched base pairs
in double-stranded DNA (so called DNA mismatch repair enzymes) in
defined systems for detecting and mapping point mutations in cDNAs
obtained from samples of cells. For example, the mutY enzyme of E.
coli cleaves following A residues at G/A mismatches and the
thymidine DNA glycosylase from HeLa cells cleaves following T
residues at G/T mismatches (Hsu et al. (1994) Carcinogenesis
15:1657-1662). According to one embodiment, a probe based on a
selected sequence, e.g., a wild-type sequence, is hybridized with a
cDNA or other DNA product obtained from a test cell(s). The duplex
is treated with a DNA mismatch repair enzyme, and the cleavage
products, if any, are detected using an electrophoresis protocol or
another polynucleotide-separating method. See, e.g., U.S. Pat. No.
5,459,039.
[0350] In other embodiments, alterations in electrophoretic
mobility are used to identify mutations in genes. For example,
single strand conformation polymorphism (SSCP) analysis can be used
to detect differences in electrophoretic mobility between mutant
and wild type nucleic acids (Orita et al. (1989) Proc. Natl. Acad.
Sci. USA 86:2766; see also Cotton (1993) Mutat. Res. 285:125-144;
Hayashi (1992) Genet. Anal. Tech. Appl. 9:73-79). Single-stranded
DNA fragments of sample and control nucleic acids are denatured and
allowed to re-nature. The secondary structure of single-stranded
nucleic acids varies according to their nucleotide sequence, and
the resulting alteration in electrophoretic mobility enables
detection of even a single base change. The DNA fragments can be
labeled or detected using labeled probes. The sensitivity of the
assay can be enhanced by using RNA (rather than DNA), because the
secondary structure of RNA is more sensitive to sequence changes.
In one embodiment, the method uses heteroduplex analysis to
separate double stranded heteroduplex molecules on the basis of
changes in electrophoretic mobility (Keen et al. (1991) Trends
Genet. 7:5).
[0351] In yet another embodiment, the movement of mutant or
wild-type fragments in polyacrylamide gels containing a gradient of
denaturant is assayed using denaturing gradient gel electrophoresis
(DGGE), as described (Myers et al. (1985) Nature 313:495). When
DGGE is used as the method of analysis, DNA is modified to ensure
that it does not completely denature, for example by adding a `GC
clamp` of approximately 40 nucleotide residues of high-melting
GC-rich DNA to one or both ends of the DNA strands, for example
using a PCR method. In a further embodiment, a temperature gradient
is used in place of a denaturing gradient to identify differences
in the mobility of control and sample DNA (Rosenbaum and Reissner
(1987) Biophys. Chem. 265:12753).
[0352] Examples of other techniques for detecting point mutations
include, but are not limited to, selective oligonucleotide
hybridization, selective amplification, and selective primer
extension. For example, oligonucleotide primers can be prepared in
which the known mutation is located centrally. The primers are
hybridized with target DNA under conditions which permit
hybridization only if a perfect complementary nucleotide sequence
match occurs (Saiki et al. (1986) Nature 324:163); Saiki et al.
(1989) Proc. Natl. Acad. Sci. USA 86:6230). Such allele specific
oligonucleotides are hybridized with PCR-amplified target DNA or
attached to a surface for hybridization.
[0353] Alternatively, allele specific amplification technology can
be used in conjunction with the methods of the invention.
Oligonucleotides used as primers for specific amplification have a
sequence complementary to the nucleotide sequence of a mutation of
interest in the center of the molecule, so that occurrence of
amplification depends on occurrence of the mutation in the sample
nucleic acid (Gibbs et al. (1989) Nucleic Acids Res. 17:2437-2448)
or at the extreme 3' end of one primer where, under appropriate
conditions, mismatching can prevent or inhibit polymerase extension
(Prossner (1993) Tibtech 11:238). In addition, it can be desirable
to introduce a novel restriction site in the region of the mutation
in order to facilitate cleavage-based detection (Gasparini et al.
(1992) Mol. Cell. Probes 6:1). Amplification can be performed using
Taq ligase (Barany (1991) Proc. Natl. Acad. Sci. USA 88:189). In
such cases, ligation will occur only if there is a perfect match at
the 3' end of the 5' sequence, thereby making it possible to assess
the presence of a known mutation at a specific site by looking for
the presence or absence of amplification.
[0354] The methods described herein can be performed, for example,
using pre-packaged diagnostic kits comprising at least one probe
nucleic acid or antibody reagent described herein. Such kits can be
used, for example, in clinical settings to diagnose patients
exhibiting symptoms or a family history of a disorder involving a
gene encoding a polypeptide of the invention. Furthermore, any cell
type or tissue in which the polypeptide of the invention is
expressed (e.g., a blood sample containing peripheral blood
leukocytes for proteins which are secreted or which occur on or in
peripheral blood leukocytes) can be used in the prognostic assays
described herein.
3. Pharmacogenomics
[0355] Agents which have a stimulatory or inhibitory effect on
activity or expression of a polypeptide of the invention, as
identified by a screening assay described herein for example, can
be administered to individuals to treat (prophylactically or
therapeutically) disorders associated with aberrant activity of the
polypeptide. In conjunction with such treatment, the
pharmacogenomics (i.e., the study of the relationship between an
individual's genotype and that individual's response to a foreign
compound or drug) of the individual can be considered. Differences
in metabolism of therapeutics can lead to severe toxicity or
therapeutic failure by altering the relation between dose and blood
concentration of the pharmacologically active drug. Thus, the
pharmacogenomics of the individual permits selection of effective
agents (e.g., drugs) for prophylactic or therapeutic treatments
based on a consideration of the individual's genotype. Such
pharmacogenomics can further be used to determine appropriate
dosages and therapeutic regimens. Accordingly, the activity of a
polypeptide of the invention, expression of a nucleic acid of the
invention, or mutation content of a gene of the invention in an
individual can be determined to facilitate selection of one or more
appropriate agents for therapeutic or prophylactic treatment of the
individual.
[0356] Pharmacogenomics deals with clinically significant
hereditary variations in the response to drugs due to altered drug
disposition and abnormal action in affected persons. See, e.g.,
Linder (1997) Clin. Chem. 43(2):254-266. In general, two types of
pharmacogenetic conditions can be differentiated. Genetic
conditions transmitted as a single factor altering the way drugs
act on the body are referred to as "altered drug action." Genetic
conditions transmitted as single factors altering the way the body
acts on drugs are referred to as "altered drug metabolism." These
pharmacogenetic conditions can occur either as rare defects or as
polymorphisms. For example, glucose-6-phosphate dehydrogenase
(G6PD) deficiency is a common inherited enzymopathy in which the
main clinical complication is hemolysis after ingestion of oxidant
drugs (anti-malarials, sulfonamides, analgesics, nitrofurans) and
consumption of fava beans.
[0357] As an illustrative embodiment, the activity of drug
metabolizing enzymes is a major determinant of both the intensity
and duration of drug action. The discovery of genetic polymorphisms
of drug metabolizing enzymes (e.g., N-acetyltransferase 2 {NAT 2}
and cytochrome P450 enzymes CYP2D6 and CYP2C19) explains why some
patients do not obtain the expected drug effects or exhibit
exaggerated drug response and serious toxicity following
administration of standard and safe doses of a drug. These
polymorphisms are expressed in two phenotypes in the population,
the extensive metabolizer (EM) and poor metabolizer (PM). The
prevalence of PM is different among different populations. For
example, the gene encoding CYP2D6 is highly polymorphic, and
several mutations have been identified in PM. Each of these
mutations results in absence of functional CYP2D6. Poor
metabolizers of CYP2D6 and CYP2C19 frequently experience
exaggerated drug response and side effects when they receive
standard doses. If a metabolite is the active therapeutic moiety, a
PM will show no therapeutic response, as demonstrated for the
analgesic effect of codeine mediated by its CYP2D6-formed
metabolite morphine. At the other extreme are the so called
ultra-rapid metabolizers who do not respond to standard doses.
Recently, the molecular basis of ultra-rapid metabolism has been
identified to be due to CYP2D6 gene amplification.
[0358] Thus, activity of a polypeptide of the invention, expression
of a nucleic acid encoding the polypeptide, or mutation content of
a gene encoding the polypeptide in an individual can be determined
to facilitate selection of appropriate agents for therapeutic or
prophylactic treatment of the individual. In addition,
pharmacogenetic studies can be used to apply genotyping of
polymorphic alleles encoding drug-metabolizing enzymes to
identification of an individual's drug responsiveness phenotype.
This knowledge, when applied to dosing or drug selection, can avoid
adverse reactions or therapeutic failure and thus enhance
therapeutic or prophylactic efficiency when treating a subject with
a modulator of activity or expression of the polypeptide, such as a
modulator identified by one of the screening assays described
herein.
4. Monitoring of Effects During Clinical Trials
[0359] Monitoring the influence of agents (e.g., drug compounds) on
expression or activity of a polypeptide of the invention (e.g.,
ability to modulate aberrant cell proliferation chemotaxis,
differentiation, or both) can be applied not only in basic drug
screening, but also in clinical trials. For example, the
effectiveness of an agent, as determined by a screening assay as
described herein, to increase gene expression, protein levels, or
protein activity can be monitored in clinical trials of subjects
exhibiting decreased gene expression, protein levels, or protein
activity. Alternatively, the effectiveness of an agent, as
determined by a screening assay, to decrease gene expression,
protein levels, or protein activity can be monitored in clinical
trials of subjects exhibiting increased gene expression, protein
levels, or protein activity. In such clinical trials, expression or
activity of a polypeptide of the invention and, optionally, that of
other polypeptide that have been implicated in similar disorders,
can be used as a marker of the immune responsiveness of a
particular cell.
[0360] For example, genes (including those of the invention) that
are modulated in cells by treatment with an agent (e.g., a peptide,
a drug, or another small molecule) which modulates activity or
expression of a polypeptide of the invention (e.g., as identified
in a screening assay described herein) can be identified. Thus, to
study the effect of agents on cellular proliferation disorders, for
example, in a clinical trial, cells can be isolated and their RNA
can be prepared and analyzed to determine the level of expression
of one or more genes of the invention and, optionally, other genes
implicated in the disorder. The levels of gene expression (i.e., a
gene expression pattern) can be quantified by Northern blot
analysis or by RT-PCR, as described herein, or by assessing the
amount of protein produced, by one of the methods as described
herein, or by measuring the level of activity of a gene of the
invention or other gene(s). In this way, the gene expression
pattern can serve as an indicator of the physiological response of
the cells to the agent. Accordingly, this response state can be
determined before, and at various points during, or after treatment
of the individual with the agent (or, of course, at more than one
of these stages).
[0361] In one embodiment, the present invention provides a method
for monitoring the effectiveness of treatment of a subject with an
agent (e.g., an agonist, antagonist, peptidomimetic, protein,
peptide, nucleic acid, small molecule, or other drug candidate
identified by the screening assays described herein) comprising (i)
obtaining a pre-administration sample from a subject prior to
administration of the agent; (ii) detecting the level of the
polypeptide or nucleic acid of the invention in the
pre-administration sample; (iii) obtaining one or more
post-administration samples from the subject; (iv) detecting the
level the of the polypeptide or nucleic acid of the invention in
the post-administration sample(s); (v) comparing the level of the
polypeptide or nucleic acid of the invention in the
pre-administration sample with the level of the polypeptide or
nucleic acid of the invention in the post-administration sample(s);
and (vi) altering the administration of the agent to the subject
accordingly. For example, increased administration of the agent can
be desirable to increase the expression or activity of the
polypeptide to levels higher than those detected, i.e., to increase
the effectiveness of the agent. Alternatively, decreased
administration of the agent can be desirable to decrease expression
or activity of the polypeptide to levels lower than those detected,
i.e., to decrease the effectiveness of the agent.
C. Methods of Treatment
[0362] The present invention provides both prophylactic and
therapeutic methods of treating a subject afflicted with, at risk
for developing, or susceptible to a disorder associated with
aberrant expression or activity of a polypeptide of the invention.
Such disorders are described elsewhere in this disclosure.
1. Prophylactic Methods
[0363] In one aspect, the invention provides a method for
preventing in a subject, a disorder associated with aberrant
expression or activity of a polypeptide of the invention, by
administering to the subject an agent which modulates expression of
the polypeptide or at least one activity of the polypeptide.
Subjects at risk for a disease which is caused or contributed to by
aberrant expression or activity of a polypeptide of the invention
can be identified by, for example, any one or combination of the
diagnostic and prognostic assays described herein. Administration
of a prophylactic agent can occur prior to the manifestation of
symptoms characteristic of the aberrance, so that the disease or
disorder is prevented or, alternatively, delayed in its onset or
progression. Depending on the type of aberrance, for example, an
agonist or antagonist agent can be used for treating the subject.
The appropriate agent can be determined based on screening assays
described herein.
2. Therapeutic Methods
[0364] Another aspect of the invention pertains to methods of
modulating expression or activity of a polypeptide of the invention
for therapeutic purposes. The modulatory method of the invention
involves contacting a cell with an agent that modulates one or more
of the activities of the polypeptide. An agent that modulates
activity can be an agent as described herein, such as a nucleic
acid, or a protein, a naturally-occurring cognate ligand of the
polypeptide, a peptide, a peptidomimetic, or a small molecule. In
one embodiment, the agent stimulates one or more of the biological
activities of the polypeptide. Examples of such stimulatory agents
include a polypeptide of the invention, a biologically active
portion of such a polypeptide, a portion of such a polypeptide
which comprises an epitope of the native polypeptide, and a nucleic
acid molecule encoding the polypeptide of the invention that has
been introduced into the cell. In another embodiment, the agent
inhibits a biological activity of the polypeptide of the invention
or expression of a protein or nucleic acid of the invention.
Examples of such inhibitory agents include antisense nucleic acid
molecules and antibodies. These modulatory methods can be performed
in vitro (e.g., by culturing the cell with the agent) or,
alternatively, in vivo (e.g., by administering the agent to a
subject). As such, the present invention provides methods of
treating an individual afflicted with a disease or disorder
characterized by aberrant expression or activity of a polypeptide
of the invention. In one embodiment, the method involves
administering an agent (e.g., an agent identified by a screening
assay described herein), or combination of agents that modulates
(e.g., up-regulates or down-regulates) expression or activity. In
another embodiment, the method involves administering a polypeptide
of the invention or a nucleic acid molecule of the invention as
therapy to compensate or substitute for reduced or aberrant
expression or activity of the polypeptide.
[0365] Stimulation of activity is desirable in situations in which
activity or expression is abnormally low or in which increased
activity is likely to have a beneficial effect. Conversely,
inhibition of activity is desirable in situations in which activity
or expression is abnormally high or in which decreased activity is
likely to have a beneficial effect.
[0366] The contents of all references, patents, and published
patent applications cited in this disclosure are hereby
incorporated by reference.
Deposits of Clones
[0367] Clone containing one or more cDNA molecules encoding
polypeptides of the invention have been deposited with the American
Type Culture Collection (ATCC.RTM.; 10801 University Boulevard,
Manassas, Va. 20110-2209) on dates disclosed herein, and these
deposits were assigned the Accession Numbers disclosed herein.
These deposits will be maintained under the terms of the Budapest
Treaty on the International Recognition of the Deposit of
Microorganisms for the Purposes of Patent Procedure. These deposits
were made merely as a convenience for those of skill in the art and
are not an admission that any deposit is required in order to
comply with 35 U.S.C. .sctn.112.
[0368] Where a clone containing multiple cDNA molecules was
deposited, the following standard digest procedure can be used to
liberate fragments corresponding to individual cDNA molecules,
except as otherwise described. To isolate the cDNA clone, an
aliquot of the deposited clone can be streaked out to yield single
colonies on nutrient medium (e.g., Luria broth plates) supplemented
with 100 micrograms per milliliter ampicillin. Single colonies are
grown, and plasmid DNA is extracted from single colonies using a
standard mini-preparation procedure. Next, a sample of the DNA
mini-preparation is digested using a combination of the restriction
enzymes Sal I and Not I, and the resulting products are resolved on
a 0.8% (w/v) agarose gel using standard DNA electrophoresis
conditions.
[0369] Clone EpT273, encoding human TANGO 273 was deposited with
ATCC.RTM. on Apr. 2, 1999 and was assigned Accession Number
207185.
[0370] Clones containing cDNA molecules encoding murine TANGO 273
were deposited with ATCC.RTM. on Apr. 21, 1999 and were assigned
Accession Number 207221, as part of a composite deposit
representing a mixture of five strains, each carrying one
recombinant plasmid harboring a particular cDNA clone. The standard
digest procedure (except that restriction enzymes SalI, NotI, and
ApaI are used) liberates a fragment as follows: [0371] mouse TANGO
273 (clone EpTm273): 0.3 kilobase and 2.6 kilobase (mouse TANGO 273
has a Apa I cut site at about base pair 298). The identity of the
strain can be inferred from the fragment liberated.
[0372] Clones comprising cDNA molecules encoding human TANGO 325
were deposited with ATCC.RTM. on May 28, 1999, as part of a
composite deposit representing a mixture of five strains, each
carrying one recombinant plasmid harboring a particular cDNA clone.
This deposit was assigned Accession Number PTA-147. The standard
digest procedure (except that restriction enzymes SalI, NotI, and
SmaI are used) liberates a fragment as follows: [0373] human TANGO
325 (clone EpT325): 2.2 kilobases The identity of the strain can be
inferred from the fragment liberated.
[0374] Clones containing cDNA molecules encoding TANGO 364 (clones
Aped), were deposited with ATCC.RTM. on Jul. 23, 1999 as Accession
No. PTA-425, as part of a composite deposit representing a mixture
of three strains, each carrying one recombinant plasmid harboring a
particular cDNA clone. The standard digest procedure liberates a
fragment as follows: [0375] TANGO 364 (Aped): 3.5 kilobase pairs
The identity of the strain can be inferred from the fragment
liberated.
[0376] Clones containing cDNA molecules encoding TANGO 405
(including clone 405), were deposited with ATCC.RTM. on Jul. 23,
1999 as Accession No. PTA-424, as part of a composite deposit
representing a mixture of five strains, each carrying one
recombinant plasmid harboring a particular cDNA clone. The standard
digest procedure liberates a fragment as follows: [0377] TANGO 405
(405): 3.1 kilobase pairs The identity of the strain can be
inferred from the fragment liberated.
EQUIVALENTS
[0378] Those skilled in the art will recognize, or be able to
ascertain using no more than routine experimentation, many
equivalents to the embodiments of the invention described herein.
Such equivalents are encompassed by the following claims.
Sequence CWU 1 SEQUENCE LISTING <160> NUMBER OF SEQ ID
NOS: 85 <210> SEQ ID NO 1 <211> LENGTH: 2964
<212> TYPE: DNA <213> ORGANISM: Homo sapiens
<400> SEQUENCE: 1 gtcgacccac gcgtccgcgg acgcgtgggg acggctcccg
gctgcagtct gcccgcccgc 60 cccgcgcggg ggccgagtcg cgaagcgcgc
ctgcgacccg gcgtccgggc gcgctggaga 120 ggacgcgagg agccatgagg
cgccagcctg cgaaggtggc ggcgctgctg ctcgggctgc 180 tcttggagtg
cacagaagcc aaaaagcatt gctggtattt cgaaggactc tatccaacct 240
attatatatg ccgctcctac gaggactgct gtggctccag gtgctgtgtg cgggccctct
300 ccatacagag gctgtggtac ttctggttcc ttctgatgat gggcgtgctt
ttctgctgcg 360 gagccggctt cttcatccgg aggcgcatgt accccccgcc
gctgatcgag gagccagcct 420 tcaatgtgtc ctacaccagg cagcccccaa
atcccggccc aggagcccag cagccggggc 480 cgccctatta cactgaccca
ggaggaccgg ggatgaaccc tgtcgggaat tccatggcaa 540 tggctttcca
ggtcccaccc aactcacccc aggggagtgt ggcctgcccg ccccctccag 600
cctactgcaa cacgcctccg cccccgtacg aacaggtagt gaaggccaag tagtggggtg
660 cccacgtgca agaggagaga caggagaggg cctttccctg gcctttctgt
cttcgttgat 720 gttcacttcc aggaacggtc tcgtgggctg ctaagggcag
ttcctctgat atcctcacag 780 caagcacagc tctctttcag gctttccatg
gagtacaata tatgaactca cactttgtct 840 cctctgttgc ttctgtttct
gacgcagtct gtgctctcac atggtagtgt ggtgacagtc 900 cccgagggct
gacgtcctta cggtggcgtg accagatcta caggagagag actgagagga 960
agaaggcagt gctggaggtg caggtggcat gtagaggggc caggccgagc atcccaggca
1020 agcatccttc tgcccgggta ttaataggaa gccccatgcc gggcggctca
gccgatgaag 1080 cagcagccga ctgagctgag cccagcaggt catctgctcc
agcctgtcct ctcgtcagcc 1140 ttcctcttcc agaagctgtt ggagagacat
tcaggagaga gcaagcccct tgtcatgttt 1200 ctgtctctgt tcatatccta
aagatagact tctcctgcac cgccagggaa gggtagcacg 1260 tgcagctctc
accgcaggat ggggcctaga atcaggcttg ccttggaggc ctgacagtga 1320
tctgacatcc actaagcaaa tttatttaaa ttcatgggaa atcacttcct gccccaaact
1380 gagacattgc attttgtgag ctcttggtct gatttggaga aaggactgtt
acccattttt 1440 ttggtgtgtt tatggaagtg catgtagagc gtcctgccct
ttgaaatcag actgggtgtg 1500 tgtcttccct ggacatcact gcctctccag
ggcattctca ggcccggggg tctccttccc 1560 tcaggcagct ccagtggtgg
gttctgaagg gtgctttcaa aacggggcac atctggctgg 1620 gaagtcacat
ggactcttcc agggagagag accagctgag gcgtctctct ctgaggttgt 1680
gttgggtcta agcgggtgtg tgctgggctc caaggaggag gagcttgctg ggaaaagaca
1740 ggagaagtac tgactcaact gcactgacca tgttgtcata attagaataa
agaagaagtg 1800 gtcggaaatg cacattcctg gataggaatc acagctcacc
ccaggatctc acaggtagtc 1860 tcctgagtag ttgacggcta gcggggagct
agttccgccg catagttata gtgttgatgt 1920 gtgaacgctg acctgtcctg
tgtgctaaga gctatgcagc ttagctgagg cgcctagatt 1980 actagatgtg
ctgtatcacg gggaatgagg tgggggtgct tattttttaa tgaactaatc 2040
agagcctctt gagaaattgt tactcattga actggagcat caagacatct catggaagtg
2100 gatacggagt gatttggtgt ccatgctttt cactctgagg acatttaatc
ggagaacctc 2160 ctggggaatt ttgtgggaga cacttgggaa caaaacagac
accctgggaa tgcagttgca 2220 agcacagatg ctgccaccag tgtctctgac
caccctggtg tgactgctga ctgccagcgt 2280 ggtacctccc atgctgcagg
cctccatcta aatgagacaa caaagcacaa tgttcactgt 2340 ttacaaccaa
gacaactgcg tgggtccaaa cactcctctt cctccaggtc atttgttttg 2400
catttttaat gtctttattt tttgtaatga aaaagcacac taagctgccc ctggaatcgg
2460 gtgcagctga ataggcaccc aaaagtccgt gactaaattt cgtttgtctt
tttgatagca 2520 aattatgtta agagacagtg atggctaggg ctcaacaatt
ttgtattccc atgtttgtgt 2580 gagacagagt ttgttttccc ttgaacttgg
ttagaattgt gctactgtga acgctgatcc 2640 tgcatatgga agtcccactt
tggtgacatt tcctggccat tcttgtttcc attgtgtgga 2700 tggtgggttg
tgcccacttc ctggagtgag acagctcctg gtgtgtagaa ttcccggagc 2760
gtccgtggtt cagagtaaac ttgaagcaga tctgtgcatg cttttcctct gcaacaattg
2820 gctcgtttct cttttttgtt ctcttttgat aggatcctgt ttcctatgtg
tgcaaaataa 2880 aaataaattt gggcaaaaaa aaaaaaaaaa aaaaaaaaaa
aaaaaaaaaa aaaaaaaaaa 2940 aaaaaaaaaa aaaagggcgg ccgc 2964
<210> SEQ ID NO 2 <211> LENGTH: 516 <212> TYPE:
DNA <213> ORGANISM: Homo sapiens <400> SEQUENCE: 2
atgaggcgcc agcctgcgaa ggtggcggcg ctgctgctcg ggctgctctt ggagtgcaca
60 gaagccaaaa agcattgctg gtatttcgaa ggactctatc caacctatta
tatatgccgc 120 tcctacgagg actgctgtgg ctccaggtgc tgtgtgcggg
ccctctccat acagaggctg 180 tggtacttct ggttccttct gatgatgggc
gtgcttttct gctgcggagc cggcttcttc 240 atccggaggc gcatgtaccc
cccgccgctg atcgaggagc cagccttcaa tgtgtcctac 300 accaggcagc
ccccaaatcc cggcccagga gcccagcagc cggggccgcc ctattacact 360
gacccaggag gaccggggat gaaccctgtc gggaattcca tggcaatggc tttccaggtc
420 ccacccaact caccccaggg gagtgtggcc tgcccgcccc ctccagccta
ctgcaacacg 480 cctccgcccc cgtacgaaca ggtagtgaag gccaag 516
<210> SEQ ID NO 3 <211> LENGTH: 172 <212> TYPE:
PRT <213> ORGANISM: Homo sapiens <400> SEQUENCE: 3 Met
Arg Arg Gln Pro Ala Lys Val Ala Ala Leu Leu Leu Gly Leu Leu 1 5 10
15 Leu Glu Cys Thr Glu Ala Lys Lys His Cys Trp Tyr Phe Glu Gly Leu
20 25 30 Tyr Pro Thr Tyr Tyr Ile Cys Arg Ser Tyr Glu Asp Cys Cys
Gly Ser 35 40 45 Arg Cys Cys Val Arg Ala Leu Ser Ile Gln Arg Leu
Trp Tyr Phe Trp 50 55 60 Phe Leu Leu Met Met Gly Val Leu Phe Cys
Cys Gly Ala Gly Phe Phe 65 70 75 80 Ile Arg Arg Arg Met Tyr Pro Pro
Pro Leu Ile Glu Glu Pro Ala Phe 85 90 95 Asn Val Ser Tyr Thr Arg
Gln Pro Pro Asn Pro Gly Pro Gly Ala Gln 100 105 110 Gln Pro Gly Pro
Pro Tyr Tyr Thr Asp Pro Gly Gly Pro Gly Met Asn 115 120 125 Pro Val
Gly Asn Ser Met Ala Met Ala Phe Gln Val Pro Pro Asn Ser 130 135 140
Pro Gln Gly Ser Val Ala Cys Pro Pro Pro Pro Ala Tyr Cys Asn Thr 145
150 155 160 Pro Pro Pro Pro Tyr Glu Gln Val Val Lys Ala Lys 165 170
<210> SEQ ID NO 4 <211> LENGTH: 22 <212> TYPE:
PRT <213> ORGANISM: Homo sapiens <400> SEQUENCE: 4 Met
Arg Arg Gln Pro Ala Lys Val Ala Ala Leu Leu Leu Gly Leu Leu 1 5 10
15 Leu Glu Cys Thr Glu Ala 20 <210> SEQ ID NO 5 <211>
LENGTH: 150 <212> TYPE: PRT <213> ORGANISM: Homo
sapiens <400> SEQUENCE: 5 Lys Lys His Cys Trp Tyr Phe Glu Gly
Leu Tyr Pro Thr Tyr Tyr Ile 1 5 10 15 Cys Arg Ser Tyr Glu Asp Cys
Cys Gly Ser Arg Cys Cys Val Arg Ala 20 25 30 Leu Ser Ile Gln Arg
Leu Trp Tyr Phe Trp Phe Leu Leu Met Met Gly 35 40 45 Val Leu Phe
Cys Cys Gly Ala Gly Phe Phe Ile Arg Arg Arg Met Tyr 50 55 60 Pro
Pro Pro Leu Ile Glu Glu Pro Ala Phe Asn Val Ser Tyr Thr Arg 65 70
75 80 Gln Pro Pro Asn Pro Gly Pro Gly Ala Gln Gln Pro Gly Pro Pro
Tyr 85 90 95 Tyr Thr Asp Pro Gly Gly Pro Gly Met Asn Pro Val Gly
Asn Ser Met 100 105 110 Ala Met Ala Phe Gln Val Pro Pro Asn Ser Pro
Gln Gly Ser Val Ala 115 120 125 Cys Pro Pro Pro Pro Ala Tyr Cys Asn
Thr Pro Pro Pro Pro Tyr Glu 130 135 140 Gln Val Val Lys Ala Lys 145
150 <210> SEQ ID NO 6 <211> LENGTH: 38 <212>
TYPE: PRT <213> ORGANISM: Homo sapiens <400> SEQUENCE:
6 Lys Lys His Cys Trp Tyr Phe Glu Gly Leu Tyr Pro Thr Tyr Tyr Ile 1
5 10 15 Cys Arg Ser Tyr Glu Asp Cys Cys Gly Ser Arg Cys Cys Val Arg
Ala 20 25 30 Leu Ser Ile Gln Arg Leu 35 <210> SEQ ID NO 7
<211> LENGTH: 21 <212> TYPE: PRT <213> ORGANISM:
Homo sapiens <400> SEQUENCE: 7 Trp Tyr Phe Trp Phe Leu Leu
Met Met Gly Val Leu Phe Cys Cys Gly 1 5 10 15 Ala Gly Phe Phe Ile
20 <210> SEQ ID NO 8 <211> LENGTH: 91 <212> TYPE:
PRT <213> ORGANISM: Homo sapiens <400> SEQUENCE: 8 Arg
Arg Arg Met Tyr Pro Pro Pro Leu Ile Glu Glu Pro Ala Phe Asn 1 5 10
15 Val Ser Tyr Thr Arg Gln Pro Pro Asn Pro Gly Pro Gly Ala Gln Gln
20 25 30 Pro Gly Pro Pro Tyr Tyr Thr Asp Pro Gly Gly Pro Gly Met
Asn Pro 35 40 45 Val Gly Asn Ser Met Ala Met Ala Phe Gln Val Pro
Pro Asn Ser Pro 50 55 60 Gln Gly Ser Val Ala Cys Pro Pro Pro Pro
Ala Tyr Cys Asn Thr Pro 65 70 75 80 Pro Pro Pro Tyr Glu Gln Val Val
Lys Ala Lys 85 90 <210> SEQ ID NO 9 <211> LENGTH:
<212> TYPE: <213> ORGANISM: <400> SEQUENCE: 9 000
<210> SEQ ID NO 10 <211> LENGTH: <212> TYPE:
<213> ORGANISM: <400> SEQUENCE: 10 000 <210> SEQ
ID NO 11 <211> LENGTH: 2915 <212> TYPE: DNA <213>
ORGANISM: Mus sp. <400> SEQUENCE: 11 gtcgacccac gcgtccggcc
gcgcgtcctt ctgccggctt cagctcgtat ccccggagtc 60 cacccgcccg
tcccggggtg cggactggcc ctgagctggc cgtacagccc ggcttcggac 120
ggtcctcgct ggagccatgg gccgccggct cggcagggtg gcggcgctgc tgctcgggct
180 gctagtggag tgcactgagg ccaaaaaaca ttgctggtat tttgaaggac
tctatcccac 240 atactatata tgccgttcct atgaagactg ctgtggctcc
aggtgctgtg tgagggccct 300 ttccatacag aggctgtggt atttttggtt
cctgctgatg atgggtgtgc tgttctgctg 360 tggtgccggt ttcttcattc
gccggcgcat gtatccgcca ccactcattg aggagcccac 420 attcaatgtg
tcctatacca ggcagccacc aaatcctgct ccaggagcac agcaaatggg 480
accgccatat tacaccgacc ctggaggacc cgggatgaat cctgttggca ataccatggc
540 tatggctttc caggtccagc ccaattcacc tcacggaggc acaacttacc
caccccctcc 600 ttcctactgc aacacgcctc caccccccta tgaacaggtg
gtgaaggaca agtagcaaga 660 tgctacatca aaggcaaaga ggatggacag
gcccttttgt ttaccttccc atcctcaccg 720 atacttgctg atagggtggt
ccaagggaaa acttggatat tctcaaagca agcccagctc 780 tctttcaagt
cttttgtgga ggacatttga atccacactg tctcctctgt tgcttctgtt 840
tctgatgtag tctgtgctct ctgagagagt gtggcaacag tccctgaggg ttgatattcc
900 tagggtgtcc agggtagatc ctcgggagag aggctaaggg gaaaggaagg
catagcctgt 960 gtgttagggg gcagataaag tggtcaggct gagataagac
tcacatgatg cagtagttgg 1020 cagtgaactt cgaagagaca ctatccacca
tcccagccca ttctcctaat agaagctgtg 1080 gggctgtgtt gttgatgctc
tttggtctcc actcacattt tgaaaatagg ctttcctctg 1140 caggaatagg
aaagacccaa gtacatattt gcttccactt aaaaatgagg gtcagaacca 1200
ggcctcagtt ggacatctat agttaaataa aggccattag agaggggaaa tctttaagtt
1260 aggggaaatt ctctaaatgg agacattgcg ttttatgaat catcgtctgg
cttttctttt 1320 agtgcatgta ttgaagtgag ggtgtccttt gagatcagat
ggggagagtg aactctgcgg 1380 ggggtggggt gtctctactc agagggctcc
aacacccttt tcttaggtag ttctggtgat 1440 gggttttatg ggcactatag
agctgagggg cacattaggc cgggtagtta cattgaccct 1500 tggagaggaa
gaggacagcc aaagaaactc agcaaagcaa gaccagcatt gctgagttag 1560
agctagggtt gtatgtgatc ccaacagaga tgtgctggcc tcagaagagg ggacgtttgt
1620 ggatagagcc gtgaaaacct acttagttgc acagatgaca taatcaaaag
tagagaaaga 1680 agtgtagtta gagatgccat ttcccaggtg agaatcagag
ctcatccata gatttacaag 1740 tagtggctgg agttaacagt atggagttct
tttcccttgc gtagttagtc acgttgatgt 1800 gtatttaaac ccaggttgag
accttgtgta ctaagagcaa ggaagtatag ctaagatgtc 1860 tagattattt
atatgtagta tggtggggag tggggctgca aggaaggggg ctgacattgt 1920
aaatgagaaa atcagagcca tttgataaac tgttacttgt tggatcaggc atccaaaagt
1980 gtctcttgag tggacattga gtattcttta ccacctacaa gaccaggagg
catggtgtca 2040 ttctccattg gggtatttat atgaggtaga ggttcaggaa
tcgacagtag ctgtgtgggc 2100 ttagtttaag gactgaaagc atagggactg
gtagacagtt tcataggaaa ctgcggggaa 2160 ggaatggata cctttaaaga
cagtttgtgg atgcagatgc tgccacccat cattgagcac 2220 ccttgtgtct
ctggcttcct gtcactggat ccagtacccc tccatgcttg ggtccttgtt 2280
ttacataaga caacaaagca caatgtctgc tgtttacaat caagacgact acatggtcca
2340 aacatttctt ctctcttcta tcacttgtgg ctttaacttc catttcctcc
gttccttttt 2400 aaaatcaaga agcacagtca gagctgcccc tgggattgca
tcagggaacg gctgatcaag 2460 gcattcagtg tccatgacta aatcttatct
ttttgatagc aaatcctttt aagaaactga 2520 acaattgcta aggctcagca
attttatact ccaatgtctg tgtaaggtaa attttgtttg 2580 ccattgagcc
cacattggaa ttccttctga cgtcaacact gacaatgcct atggaaattg 2640
cacttctggg tatatgtccc agcatccttg ttttcttatg tttggtgagt aaggctcacc
2700 ccttccagca gctctacttc tgtgtgctga ggtcctgtag agccggggct
tgggcacaga 2760 catgaggcag acttgtgcat gctctttctt ggcaacactt
ggctcatatt tcttgttctc 2820 ttttgataga gtcctgtttc ctatgtattt
aaaaaataat aaaagtgaat ttagtcaaaa 2880 aaaaaaaaaa aaaaaaaaaa
aaaaagggcg gccgc 2915 <210> SEQ ID NO 12 <211> LENGTH:
516 <212> TYPE: DNA <213> ORGANISM: Mus sp. <400>
SEQUENCE: 12 atgggccgcc ggctcggcag ggtggcggcg ctgctgctcg ggctgctagt
ggagtgcact 60 gaggccaaaa aacattgctg gtattttgaa ggactctatc
ccacatacta tatatgccgt 120 tcctatgaag actgctgtgg ctccaggtgc
tgtgtgaggg ccctttccat acagaggctg 180 tggtattttt ggttcctgct
gatgatgggt gtgctgttct gctgtggtgc cggtttcttc 240 attcgccggc
gcatgtatcc gccaccactc attgaggagc ccacattcaa tgtgtcctat 300
accaggcagc caccaaatcc tgctccagga gcacagcaaa tgggaccgcc atattacacc
360 gaccctggag gacccgggat gaatcctgtt ggcaatacca tggctatggc
tttccaggtc 420 cagcccaatt cacctcacgg aggcacaact tacccacccc
ctccttccta ctgcaacacg 480 cctccacccc cctatgaaca ggtggtgaag gacaag
516 <210> SEQ ID NO 13 <211> LENGTH: 172 <212>
TYPE: PRT <213> ORGANISM: Mus sp. <400> SEQUENCE: 13
Met Gly Arg Arg Leu Gly Arg Val Ala Ala Leu Leu Leu Gly Leu Leu 1 5
10 15 Val Glu Cys Thr Glu Ala Lys Lys His Cys Trp Tyr Phe Glu Gly
Leu 20 25 30 Tyr Pro Thr Tyr Tyr Ile Cys Arg Ser Tyr Glu Asp Cys
Cys Gly Ser 35 40 45 Arg Cys Cys Val Arg Ala Leu Ser Ile Gln Arg
Leu Trp Tyr Phe Trp 50 55 60 Phe Leu Leu Met Met Gly Val Leu Phe
Cys Cys Gly Ala Gly Phe Phe 65 70 75 80 Ile Arg Arg Arg Met Tyr Pro
Pro Pro Leu Ile Glu Glu Pro Thr Phe 85 90 95 Asn Val Ser Tyr Thr
Arg Gln Pro Pro Asn Pro Ala Pro Gly Ala Gln 100 105 110 Gln Met Gly
Pro Pro Tyr Tyr Thr Asp Pro Gly Gly Pro Gly Met Asn 115 120 125 Pro
Val Gly Asn Thr Met Ala Met Ala Phe Gln Val Gln Pro Asn Ser 130 135
140 Pro His Gly Gly Thr Thr Tyr Pro Pro Pro Pro Ser Tyr Cys Asn Thr
145 150 155 160 Pro Pro Pro Pro Tyr Glu Gln Val Val Lys Asp Lys 165
170 <210> SEQ ID NO 14 <211> LENGTH: <212> TYPE:
<213> ORGANISM: <400> SEQUENCE: 14 000 <210> SEQ
ID NO 15 <211> LENGTH: 150 <212> TYPE: PRT <213>
ORGANISM: Mus sp. <400> SEQUENCE: 15 Lys Lys His Cys Trp Tyr
Phe Glu Gly Leu Tyr Pro Thr Tyr Tyr Ile 1 5 10 15 Cys Arg Ser Tyr
Glu Asp Cys Cys Gly Ser Arg Cys Cys Val Arg Ala 20 25 30 Leu Ser
Ile Gln Arg Leu Trp Tyr Phe Trp Phe Leu Leu Met Met Gly 35 40 45
Val Leu Phe Cys Cys Gly Ala Gly Phe Phe Ile Arg Arg Arg Met Tyr 50
55 60 Pro Pro Pro Leu Ile Glu Glu Pro Thr Phe Asn Val Ser Tyr Thr
Arg 65 70 75 80 Gln Pro Pro Asn Pro Ala Pro Gly Ala Gln Gln Met Gly
Pro Pro Tyr 85 90 95 Tyr Thr Asp Pro Gly Gly Pro Gly Met Asn Pro
Val Gly Asn Thr Met 100 105 110 Ala Met Ala Phe Gln Val Gln Pro Asn
Ser Pro His Gly Gly Thr Thr 115 120 125 Tyr Pro Pro Pro Pro Ser Tyr
Cys Asn Thr Pro Pro Pro Pro Tyr Glu 130 135 140 Gln Val Val Lys Asp
Lys 145 150 <210> SEQ ID NO 16 <400> SEQUENCE: 16 000 3
<210> SEQ ID NO 17 <400> SEQUENCE: 17 000 3 <210>
SEQ ID NO 18 <400> SEQUENCE: 18 000 3 <210> SEQ ID NO
19 <211> LENGTH: <212> TYPE: <213> ORGANISM:
<400> SEQUENCE: 19 000 <210> SEQ ID NO 20 <400>
SEQUENCE: 20 000 3 <210> SEQ ID NO 21 <211> LENGTH:
2169 <212> TYPE: DNA <213> ORGANISM: Homo sapiens
<400> SEQUENCE: 21 gtcgacccac gcgtccggaa atgtcgttct
tcagatttaa aaagaaaacc tttactgaat 60 cagctgagtg ttaataatac
gaatttcctt ttcttgccaa ttctgatctg aacagaaaat 120 ccaagaacag
ggatatgtgt ggattacagt tttctctgcc ttgcctacga ctgtttctgg 180
ttgttacctg ttatctttta ttattactcc acaaagaaat acttggatgt tcgtctgttt
240 gtcagctctg cactgggaga caaattaact gccgtaactt aggcctttcg
agtattccta 300 agaattttcc tgaaagtaca gtttttctgt atctgactgg
gaataatata tcttatataa 360 atgaaagtga attaacagga cttcattctc
ttgtagcatt gtatttggat aattctaaca 420 ttctgtatgt atatccaaaa
gcctttgttc aattgaggca tctatatttt ctatttctaa 480 ataataattt
catcaaacgc ttagatcctg gaatatttaa gggactttta aatcttcgta 540
atttatattt acagtataat caggtatctt ttgttccgag aggagtattt aatgatctag
600 tttcagttca gtacttaaat ctacaaagga atcgcctcac tgtccttggg
agtggtacct 660 ttgttggtat ggttgctctt cggatacttg atttatcaaa
caataacatt ttgaggatat 720 cagaatcagg ctttcaacat cttgaaaacc
ttgcttgttt gtatttagga agtaataatt 780 taacaaaagt accatcaaat
gcctttgaag tacttaaaag tcttagaaga ctttctttgt 840 ctcataatcc
tattgaagca atacagccct ttgcatttaa aggacttgcc aatctggaat 900
acctcctcct gaaaaattca agaattagga atgttactag ggatgggttt agtggaatta
960 ataatcttaa acatttgatc ttaagtcata atgatttaga gaatttaaat
tctgacacat 1020 tcagtttgtt aaagaattta atttacctta agttagatag
aaacagaata attagcattg 1080 ataatgatac atttgaaaat atgggagcat
ctttgaagat ccttaatctg tcatttaata 1140 atcttacagc cttgcatcca
agggtcctta agccgttgtc ttcattgatt catcttcagg 1200 caaattctaa
tccttgggaa tgtaactgca aacttttggg ccttcgagac tggctagcat 1260
cttcagccat tactctaaac atctattgtc agaatccccc atccatgcgt ggcagagcat
1320 tacgttatat taacattaca aattgtgtta catcttcaat aaatgtatcc
agagcttggg 1380 ctgttgtaaa atctcctcat attcatcaca agactactgc
gctaatgatg gcctggcata 1440 aagtaaccac aaatggcagt cctctggaaa
atactgagac tgagaacatt actttctggg 1500 aacgaattcc tacttcacct
gctggtagat tttttcaaga gaatgccttt ggtaatccat 1560 tagagactac
agcagtgtta cctgtgcaaa tacaacttac tacttctgtt accttgaact 1620
tggaaaaaaa cagtgctcta ccgaatgatg ctgcttcaat gtcagggaaa acatctctaa
1680 tttgtacaca agaagttgag aagttgaatg aggcttttga cattttgcta
gcttttttca 1740 tcttagcttg tgttttaatc atttttttga tctacaaagt
tgttcagttt aaacaaaaac 1800 taaaggcatc agaaaactca agggaaaata
gacttgaata ctacagcttt tatcagtcag 1860 caaggtataa tgtaactgcc
tcaatttgta acacttcccc aaattctcta gaaagtcctg 1920 gcttggagca
gattcgactt cataaacaaa ttgttcctga aaatgaggca caggtcattc 1980
tttttgaaca ttctgcttta taactcaact aaatattgtc tataagaaac ttcagtgcca
2040 tggacatgat ttaaactgaa acctccttat ataattatat actttagttg
gaaatataat 2100 gaattatatg aggttagcat tattaaaata tgtttttaat
aaaaaaaaaa aaaaaaaaag 2160 ggcggccgc 2169 <210> SEQ ID NO 22
<211> LENGTH: 1866 <212> TYPE: DNA <213>
ORGANISM: Homo sapiens <400> SEQUENCE: 22 atgtgtggat
tacagttttc tctgccttgc ctacgactgt ttctggttgt tacctgttat 60
cttttattat tactccacaa agaaatactt ggatgttcgt ctgtttgtca gctctgcact
120 gggagacaaa ttaactgccg taacttaggc ctttcgagta ttcctaagaa
ttttcctgaa 180 agtacagttt ttctgtatct gactgggaat aatatatctt
atataaatga aagtgaatta 240 acaggacttc attctcttgt agcattgtat
ttggataatt ctaacattct gtatgtatat 300 ccaaaagcct ttgttcaatt
gaggcatcta tattttctat ttctaaataa taatttcatc 360 aaacgcttag
atcctggaat atttaaggga cttttaaatc ttcgtaattt atatttacag 420
tataatcagg tatcttttgt tccgagagga gtatttaatg atctagtttc agttcagtac
480 ttaaatctac aaaggaatcg cctcactgtc cttgggagtg gtacctttgt
tggtatggtt 540 gctcttcgga tacttgattt atcaaacaat aacattttga
ggatatcaga atcaggcttt 600 caacatcttg aaaaccttgc ttgtttgtat
ttaggaagta ataatttaac aaaagtacca 660 tcaaatgcct ttgaagtact
taaaagtctt agaagacttt ctttgtctca taatcctatt 720 gaagcaatac
agccctttgc atttaaagga cttgccaatc tggaatacct cctcctgaaa 780
aattcaagaa ttaggaatgt tactagggat gggtttagtg gaattaataa tcttaaacat
840 ttgatcttaa gtcataatga tttagagaat ttaaattctg acacattcag
tttgttaaag 900 aatttaattt accttaagtt agatagaaac agaataatta
gcattgataa tgatacattt 960 gaaaatatgg gagcatcttt gaagatcctt
aatctgtcat ttaataatct tacagccttg 1020 catccaaggg tccttaagcc
gttgtcttca ttgattcatc ttcaggcaaa ttctaatcct 1080 tgggaatgta
actgcaaact tttgggcctt cgagactggc tagcatcttc agccattact 1140
ctaaacatct attgtcagaa tcccccatcc atgcgtggca gagcattacg ttatattaac
1200 attacaaatt gtgttacatc ttcaataaat gtatccagag cttgggctgt
tgtaaaatct 1260 cctcatattc atcacaagac tactgcgcta atgatggcct
ggcataaagt aaccacaaat 1320 ggcagtcctc tggaaaatac tgagactgag
aacattactt tctgggaacg aattcctact 1380 tcacctgctg gtagattttt
tcaagagaat gcctttggta atccattaga gactacagca 1440 gtgttacctg
tgcaaataca acttactact tctgttacct tgaacttgga aaaaaacagt 1500
gctctaccga atgatgctgc ttcaatgtca gggaaaacat ctctaatttg tacacaagaa
1560 gttgagaagt tgaatgaggc ttttgacatt ttgctagctt ttttcatctt
agcttgtgtt 1620 ttaatcattt ttttgatcta caaagttgtt cagtttaaac
aaaaactaaa ggcatcagaa 1680 aactcaaggg aaaatagact tgaatactac
agcttttatc agtcagcaag gtataatgta 1740 actgcctcaa tttgtaacac
ttccccaaat tctctagaaa gtcctggctt ggagcagatt 1800 cgacttcata
aacaaattgt tcctgaaaat gaggcacagg tcattctttt tgaacattct 1860 gcttta
1866 <210> SEQ ID NO 23 <211> LENGTH: 622 <212>
TYPE: PRT <213> ORGANISM: Homo sapiens <400> SEQUENCE:
23 Met Cys Gly Leu Gln Phe Ser Leu Pro Cys Leu Arg Leu Phe Leu Val
1 5 10 15 Val Thr Cys Tyr Leu Leu Leu Leu Leu His Lys Glu Ile Leu
Gly Cys 20 25 30 Ser Ser Val Cys Gln Leu Cys Thr Gly Arg Gln Ile
Asn Cys Arg Asn 35 40 45 Leu Gly Leu Ser Ser Ile Pro Lys Asn Phe
Pro Glu Ser Thr Val Phe 50 55 60 Leu Tyr Leu Thr Gly Asn Asn Ile
Ser Tyr Ile Asn Glu Ser Glu Leu 65 70 75 80 Thr Gly Leu His Ser Leu
Val Ala Leu Tyr Leu Asp Asn Ser Asn Ile 85 90 95 Leu Tyr Val Tyr
Pro Lys Ala Phe Val Gln Leu Arg His Leu Tyr Phe 100 105 110 Leu Phe
Leu Asn Asn Asn Phe Ile Lys Arg Leu Asp Pro Gly Ile Phe 115 120 125
Lys Gly Leu Leu Asn Leu Arg Asn Leu Tyr Leu Gln Tyr Asn Gln Val 130
135 140 Ser Phe Val Pro Arg Gly Val Phe Asn Asp Leu Val Ser Val Gln
Tyr 145 150 155 160 Leu Asn Leu Gln Arg Asn Arg Leu Thr Val Leu Gly
Ser Gly Thr Phe 165 170 175 Val Gly Met Val Ala Leu Arg Ile Leu Asp
Leu Ser Asn Asn Asn Ile 180 185 190 Leu Arg Ile Ser Glu Ser Gly Phe
Gln His Leu Glu Asn Leu Ala Cys 195 200 205 Leu Tyr Leu Gly Ser Asn
Asn Leu Thr Lys Val Pro Ser Asn Ala Phe 210 215 220 Glu Val Leu Lys
Ser Leu Arg Arg Leu Ser Leu Ser His Asn Pro Ile 225 230 235 240 Glu
Ala Ile Gln Pro Phe Ala Phe Lys Gly Leu Ala Asn Leu Glu Tyr 245 250
255 Leu Leu Leu Lys Asn Ser Arg Ile Arg Asn Val Thr Arg Asp Gly Phe
260 265 270 Ser Gly Ile Asn Asn Leu Lys His Leu Ile Leu Ser His Asn
Asp Leu 275 280 285 Glu Asn Leu Asn Ser Asp Thr Phe Ser Leu Leu Lys
Asn Leu Ile Tyr 290 295 300 Leu Lys Leu Asp Arg Asn Arg Ile Ile Ser
Ile Asp Asn Asp Thr Phe 305 310 315 320 Glu Asn Met Gly Ala Ser Leu
Lys Ile Leu Asn Leu Ser Phe Asn Asn 325 330 335 Leu Thr Ala Leu His
Pro Arg Val Leu Lys Pro Leu Ser Ser Leu Ile 340 345 350 His Leu Gln
Ala Asn Ser Asn Pro Trp Glu Cys Asn Cys Lys Leu Leu 355 360 365 Gly
Leu Arg Asp Trp Leu Ala Ser Ser Ala Ile Thr Leu Asn Ile Tyr 370 375
380 Cys Gln Asn Pro Pro Ser Met Arg Gly Arg Ala Leu Arg Tyr Ile Asn
385 390 395 400 Ile Thr Asn Cys Val Thr Ser Ser Ile Asn Val Ser Arg
Ala Trp Ala 405 410 415 Val Val Lys Ser Pro His Ile His His Lys Thr
Thr Ala Leu Met Met 420 425 430 Ala Trp His Lys Val Thr Thr Asn Gly
Ser Pro Leu Glu Asn Thr Glu 435 440 445 Thr Glu Asn Ile Thr Phe Trp
Glu Arg Ile Pro Thr Ser Pro Ala Gly 450 455 460 Arg Phe Phe Gln Glu
Asn Ala Phe Gly Asn Pro Leu Glu Thr Thr Ala 465 470 475 480 Val Leu
Pro Val Gln Ile Gln Leu Thr Thr Ser Val Thr Leu Asn Leu 485 490 495
Glu Lys Asn Ser Ala Leu Pro Asn Asp Ala Ala Ser Met Ser Gly Lys 500
505 510 Thr Ser Leu Ile Cys Thr Gln Glu Val Glu Lys Leu Asn Glu Ala
Phe 515 520 525 Asp Ile Leu Leu Ala Phe Phe Ile Leu Ala Cys Val Leu
Ile Ile Phe 530 535 540 Leu Ile Tyr Lys Val Val Gln Phe Lys Gln Lys
Leu Lys Ala Ser Glu 545 550 555 560 Asn Ser Arg Glu Asn Arg Leu Glu
Tyr Tyr Ser Phe Tyr Gln Ser Ala 565 570 575 Arg Tyr Asn Val Thr Ala
Ser Ile Cys Asn Thr Ser Pro Asn Ser Leu 580 585 590 Glu Ser Pro Gly
Leu Glu Gln Ile Arg Leu His Lys Gln Ile Val Pro 595 600 605 Glu Asn
Glu Ala Gln Val Ile Leu Phe Glu His Ser Ala Leu 610 615 620
<210> SEQ ID NO 24 <211> LENGTH: 31 <212> TYPE:
PRT <213> ORGANISM: Homo sapiens <400> SEQUENCE: 24 Met
Cys Gly Leu Gln Phe Ser Leu Pro Cys Leu Arg Leu Phe Leu Val 1 5 10
15 Val Thr Cys Tyr Leu Leu Leu Leu Leu His Lys Glu Ile Leu Gly 20
25 30 <210> SEQ ID NO 25 <211> LENGTH: 591 <212>
TYPE: PRT <213> ORGANISM: Homo sapiens <400> SEQUENCE:
25 Cys Ser Ser Val Cys Gln Leu Cys Thr Gly Arg Gln Ile Asn Cys Arg
1 5 10 15 Asn Leu Gly Leu Ser Ser Ile Pro Lys Asn Phe Pro Glu Ser
Thr Val 20 25 30 Phe Leu Tyr Leu Thr Gly Asn Asn Ile Ser Tyr Ile
Asn Glu Ser Glu 35 40 45 Leu Thr Gly Leu His Ser Leu Val Ala Leu
Tyr Leu Asp Asn Ser Asn 50 55 60 Ile Leu Tyr Val Tyr Pro Lys Ala
Phe Val Gln Leu Arg His Leu Tyr 65 70 75 80 Phe Leu Phe Leu Asn Asn
Asn Phe Ile Lys Arg Leu Asp Pro Gly Ile 85 90 95 Phe Lys Gly Leu
Leu Asn Leu Arg Asn Leu Tyr Leu Gln Tyr Asn Gln 100 105 110 Val Ser
Phe Val Pro Arg Gly Val Phe Asn Asp Leu Val Ser Val Gln 115 120 125
Tyr Leu Asn Leu Gln Arg Asn Arg Leu Thr Val Leu Gly Ser Gly Thr 130
135 140 Phe Val Gly Met Val Ala Leu Arg Ile Leu Asp Leu Ser Asn Asn
Asn 145 150 155 160 Ile Leu Arg Ile Ser Glu Ser Gly Phe Gln His Leu
Glu Asn Leu Ala 165 170 175 Cys Leu Tyr Leu Gly Ser Asn Asn Leu Thr
Lys Val Pro Ser Asn Ala 180 185 190 Phe Glu Val Leu Lys Ser Leu Arg
Arg Leu Ser Leu Ser His Asn Pro 195 200 205 Ile Glu Ala Ile Gln Pro
Phe Ala Phe Lys Gly Leu Ala Asn Leu Glu 210 215 220 Tyr Leu Leu Leu
Lys Asn Ser Arg Ile Arg Asn Val Thr Arg Asp Gly 225 230 235 240 Phe
Ser Gly Ile Asn Asn Leu Lys His Leu Ile Leu Ser His Asn Asp 245 250
255 Leu Glu Asn Leu Asn Ser Asp Thr Phe Ser Leu Leu Lys Asn Leu Ile
260 265 270 Tyr Leu Lys Leu Asp Arg Asn Arg Ile Ile Ser Ile Asp Asn
Asp Thr 275 280 285 Phe Glu Asn Met Gly Ala Ser Leu Lys Ile Leu Asn
Leu Ser Phe Asn 290 295 300 Asn Leu Thr Ala Leu His Pro Arg Val Leu
Lys Pro Leu Ser Ser Leu 305 310 315 320 Ile His Leu Gln Ala Asn Ser
Asn Pro Trp Glu Cys Asn Cys Lys Leu 325 330 335 Leu Gly Leu Arg Asp
Trp Leu Ala Ser Ser Ala Ile Thr Leu Asn Ile 340 345 350 Tyr Cys Gln
Asn Pro Pro Ser Met Arg Gly Arg Ala Leu Arg Tyr Ile 355 360 365 Asn
Ile Thr Asn Cys Val Thr Ser Ser Ile Asn Val Ser Arg Ala Trp 370 375
380 Ala Val Val Lys Ser Pro His Ile His His Lys Thr Thr Ala Leu Met
385 390 395 400 Met Ala Trp His Lys Val Thr Thr Asn Gly Ser Pro Leu
Glu Asn Thr 405 410 415 Glu Thr Glu Asn Ile Thr Phe Trp Glu Arg Ile
Pro Thr Ser Pro Ala 420 425 430 Gly Arg Phe Phe Gln Glu Asn Ala Phe
Gly Asn Pro Leu Glu Thr Thr 435 440 445 Ala Val Leu Pro Val Gln Ile
Gln Leu Thr Thr Ser Val Thr Leu Asn 450 455 460 Leu Glu Lys Asn Ser
Ala Leu Pro Asn Asp Ala Ala Ser Met Ser Gly 465 470 475 480 Lys Thr
Ser Leu Ile Cys Thr Gln Glu Val Glu Lys Leu Asn Glu Ala 485 490 495
Phe Asp Ile Leu Leu Ala Phe Phe Ile Leu Ala Cys Val Leu Ile Ile 500
505 510 Phe Leu Ile Tyr Lys Val Val Gln Phe Lys Gln Lys Leu Lys Ala
Ser 515 520 525 Glu Asn Ser Arg Glu Asn Arg Leu Glu Tyr Tyr Ser Phe
Tyr Gln Ser 530 535 540 Ala Arg Tyr Asn Val Thr Ala Ser Ile Cys Asn
Thr Ser Pro Asn Ser 545 550 555 560 Leu Glu Ser Pro Gly Leu Glu Gln
Ile Arg Leu His Lys Gln Ile Val 565 570 575 Pro Glu Asn Glu Ala Gln
Val Ile Leu Phe Glu His Ser Ala Leu 580 585 590 <210> SEQ ID
NO 26 <211> LENGTH: 498 <212> TYPE: PRT <213>
ORGANISM: Homo sapiens <400> SEQUENCE: 26 Cys Ser Ser Val Cys
Gln Leu Cys Thr Gly Arg Gln Ile Asn Cys Arg 1 5 10 15 Asn Leu Gly
Leu Ser Ser Ile Pro Lys Asn Phe Pro Glu Ser Thr Val 20 25 30 Phe
Leu Tyr Leu Thr Gly Asn Asn Ile Ser Tyr Ile Asn Glu Ser Glu 35 40
45 Leu Thr Gly Leu His Ser Leu Val Ala Leu Tyr Leu Asp Asn Ser Asn
50 55 60 Ile Leu Tyr Val Tyr Pro Lys Ala Phe Val Gln Leu Arg His
Leu Tyr 65 70 75 80 Phe Leu Phe Leu Asn Asn Asn Phe Ile Lys Arg Leu
Asp Pro Gly Ile 85 90 95 Phe Lys Gly Leu Leu Asn Leu Arg Asn Leu
Tyr Leu Gln Tyr Asn Gln 100 105 110 Val Ser Phe Val Pro Arg Gly Val
Phe Asn Asp Leu Val Ser Val Gln 115 120 125 Tyr Leu Asn Leu Gln Arg
Asn Arg Leu Thr Val Leu Gly Ser Gly Thr 130 135 140 Phe Val Gly Met
Val Ala Leu Arg Ile Leu Asp Leu Ser Asn Asn Asn 145 150 155 160 Ile
Leu Arg Ile Ser Glu Ser Gly Phe Gln His Leu Glu Asn Leu Ala 165 170
175 Cys Leu Tyr Leu Gly Ser Asn Asn Leu Thr Lys Val Pro Ser Asn Ala
180 185 190 Phe Glu Val Leu Lys Ser Leu Arg Arg Leu Ser Leu Ser His
Asn Pro 195 200 205 Ile Glu Ala Ile Gln Pro Phe Ala Phe Lys Gly Leu
Ala Asn Leu Glu 210 215 220 Tyr Leu Leu Leu Lys Asn Ser Arg Ile Arg
Asn Val Thr Arg Asp Gly 225 230 235 240 Phe Ser Gly Ile Asn Asn Leu
Lys His Leu Ile Leu Ser His Asn Asp 245 250 255 Leu Glu Asn Leu Asn
Ser Asp Thr Phe Ser Leu Leu Lys Asn Leu Ile 260 265 270 Tyr Leu Lys
Leu Asp Arg Asn Arg Ile Ile Ser Ile Asp Asn Asp Thr 275 280 285 Phe
Glu Asn Met Gly Ala Ser Leu Lys Ile Leu Asn Leu Ser Phe Asn 290 295
300 Asn Leu Thr Ala Leu His Pro Arg Val Leu Lys Pro Leu Ser Ser Leu
305 310 315 320 Ile His Leu Gln Ala Asn Ser Asn Pro Trp Glu Cys Asn
Cys Lys Leu 325 330 335 Leu Gly Leu Arg Asp Trp Leu Ala Ser Ser Ala
Ile Thr Leu Asn Ile 340 345 350 Tyr Cys Gln Asn Pro Pro Ser Met Arg
Gly Arg Ala Leu Arg Tyr Ile 355 360 365 Asn Ile Thr Asn Cys Val Thr
Ser Ser Ile Asn Val Ser Arg Ala Trp 370 375 380 Ala Val Val Lys Ser
Pro His Ile His His Lys Thr Thr Ala Leu Met 385 390 395 400 Met Ala
Trp His Lys Val Thr Thr Asn Gly Ser Pro Leu Glu Asn Thr 405 410 415
Glu Thr Glu Asn Ile Thr Phe Trp Glu Arg Ile Pro Thr Ser Pro Ala 420
425 430 Gly Arg Phe Phe Gln Glu Asn Ala Phe Gly Asn Pro Leu Glu Thr
Thr 435 440 445 Ala Val Leu Pro Val Gln Ile Gln Leu Thr Thr Ser Val
Thr Leu Asn 450 455 460 Leu Glu Lys Asn Ser Ala Leu Pro Asn Asp Ala
Ala Ser Met Ser Gly 465 470 475 480 Lys Thr Ser Leu Ile Cys Thr Gln
Glu Val Glu Lys Leu Asn Glu Ala 485 490 495 Phe Asp <210> SEQ
ID NO 27 <211> LENGTH: 18 <212> TYPE: PRT <213>
ORGANISM: Homo sapiens <400> SEQUENCE: 27 Ile Leu Leu Ala Phe
Phe Ile Leu Ala Cys Val Leu Ile Ile Phe Leu 1 5 10 15 Ile Tyr
<210> SEQ ID NO 28 <211> LENGTH: 75 <212> TYPE:
PRT <213> ORGANISM: Homo sapiens <400> SEQUENCE: 28 Lys
Val Val Gln Phe Lys Gln Lys Leu Lys Ala Ser Glu Asn Ser Arg 1 5 10
15 Glu Asn Arg Leu Glu Tyr Tyr Ser Phe Tyr Gln Ser Ala Arg Tyr Asn
20 25 30 Val Thr Ala Ser Ile Cys Asn Thr Ser Pro Asn Ser Leu Glu
Ser Pro 35 40 45 Gly Leu Glu Gln Ile Arg Leu His Lys Gln Ile Val
Pro Glu Asn Glu 50 55 60 Ala Gln Val Ile Leu Phe Glu His Ser Ala
Leu 65 70 75 <210> SEQ ID NO 29 <211> LENGTH: 1529
<212> TYPE: PRT <213> ORGANISM: Homo sapiens
<400> SEQUENCE: 29 Met Arg Gly Val Gly Trp Gln Met Leu Ser
Leu Ser Leu Gly Leu Val 1 5 10 15 Leu Ala Ile Leu Asn Lys Val Ala
Pro Gln Ala Cys Pro Ala Gln Cys 20 25 30 Ser Cys Ser Gly Ser Thr
Val Asp Cys His Gly Leu Ala Leu Arg Ser 35 40 45 Val Pro Arg Asn
Ile Pro Arg Asn Thr Glu Arg Leu Asp Leu Asn Gly 50 55 60 Asn Asn
Ile Thr Arg Ile Thr Lys Thr Asp Phe Ala Gly Leu Arg His 65 70 75 80
Leu Arg Val Leu Gln Leu Met Glu Asn Lys Ile Ser Thr Ile Glu Arg 85
90 95 Gly Ala Phe Gln Asp Leu Lys Glu Leu Glu Arg Leu Arg Leu Asn
Arg 100 105 110 Asn His Leu Gln Leu Phe Pro Glu Leu Leu Phe Leu Gly
Thr Ala Lys 115 120 125 Leu Tyr Arg Leu Asp Leu Ser Glu Asn Gln Ile
Gln Ala Ile Pro Arg 130 135 140 Lys Ala Phe Arg Gly Ala Val Asp Ile
Lys Asn Leu Gln Leu Asp Tyr 145 150 155 160 Asn Gln Ile Ser Cys Ile
Glu Asp Gly Ala Phe Arg Ala Leu Arg Asp 165 170 175 Leu Glu Val Leu
Thr Leu Asn Asn Asn Asn Ile Thr Arg Leu Ser Val 180 185 190 Ala Ser
Phe Asn His Met Pro Lys Leu Arg Thr Phe Arg Leu His Ser 195 200 205
Asn Asn Leu Tyr Cys Asp Cys His Leu Ala Trp Leu Ser Asp Trp Leu 210
215 220 Arg Gln Arg Pro Arg Val Gly Leu Tyr Thr Gln Cys Met Gly Pro
Ser 225 230 235 240 His Leu Arg Gly His Asn Val Ala Glu Val Gln Lys
Arg Glu Phe Val 245 250 255 Cys Ser Gly His Gln Ser Phe Met Ala Pro
Ser Cys Ser Val Leu His 260 265 270 Cys Pro Ala Ala Cys Thr Cys Ser
Asn Asn Ile Val Asp Cys Arg Gly 275 280 285 Lys Gly Leu Thr Glu Ile
Pro Thr Asn Leu Pro Glu Thr Ile Thr Glu 290 295 300 Ile Arg Leu Glu
Gln Asn Thr Ile Lys Val Ile Pro Pro Gly Ala Phe 305 310 315 320 Ser
Pro Tyr Lys Lys Leu Arg Arg Ile Asp Leu Ser Asn Asn Gln Ile 325 330
335 Ser Glu Leu Ala Pro Asp Ala Phe Gln Gly Leu Arg Ser Leu Asn Ser
340 345 350 Leu Val Leu Tyr Gly Asn Lys Ile Thr Glu Leu Pro Lys Ser
Leu Phe 355 360 365 Glu Gly Leu Phe Ser Leu Gln Leu Leu Leu Leu Asn
Ala Asn Lys Ile 370 375 380 Asn Cys Leu Arg Val Asp Ala Phe Gln Asp
Leu His Asn Leu Asn Leu 385 390 395 400 Leu Ser Leu Tyr Asp Asn Lys
Leu Gln Thr Ile Ala Lys Gly Thr Phe 405 410 415 Ser Pro Leu Arg Ala
Ile Gln Thr Met His Leu Ala Gln Asn Pro Phe 420 425 430 Ile Cys Asp
Cys His Leu Lys Trp Leu Ala Asp Tyr Leu His Thr Asn 435 440 445 Pro
Ile Glu Thr Ser Gly Ala Arg Cys Thr Ser Pro Arg Arg Leu Ala 450 455
460 Asn Lys Arg Ile Gly Gln Ile Lys Ser Lys Lys Phe Arg Cys Ser Ala
465 470 475 480 Lys Glu Gln Tyr Phe Ile Pro Gly Thr Glu Asp Tyr Arg
Ser Lys Leu 485 490 495 Ser Gly Asp Cys Phe Ala Asp Leu Ala Cys Pro
Glu Lys Cys Arg Cys 500 505 510 Glu Gly Thr Thr Val Asp Cys Ser Asn
Gln Lys Leu Asn Lys Ile Pro 515 520 525 Glu His Ile Pro Gln Tyr Thr
Ala Glu Leu Arg Leu Asn Asn Asn Glu 530 535 540 Phe Thr Val Leu Glu
Ala Thr Gly Ile Phe Lys Lys Leu Pro Gln Leu 545 550 555 560 Arg Lys
Ile Asn Phe Ser Asn Asn Lys Ile Thr Asp Ile Glu Glu Gly 565 570 575
Ala Phe Glu Gly Ala Ser Gly Val Asn Glu Ile Leu Leu Thr Ser Asn 580
585 590 Arg Leu Glu Asn Val Gln His Lys Met Phe Lys Gly Leu Glu Ser
Leu 595 600 605 Lys Thr Leu Met Leu Arg Ser Asn Arg Ile Thr Cys Val
Gly Asn Asp 610 615 620 Ser Phe Ile Gly Leu Ser Ser Val Arg Leu Leu
Ser Leu Tyr Asp Asn 625 630 635 640 Gln Ile Thr Thr Val Ala Pro Gly
Ala Phe Asp Thr Leu His Ser Leu 645 650 655 Ser Thr Leu Asn Leu Leu
Ala Asn Pro Phe Asn Cys Asn Cys Tyr Leu 660 665 670 Ala Trp Leu Gly
Glu Trp Leu Arg Lys Lys Arg Ile Val Thr Gly Asn 675 680 685 Pro Arg
Cys Gln Lys Pro Tyr Phe Leu Lys Glu Ile Pro Ile Gln Asp 690 695 700
Val Ala Ile Gln Asp Phe Thr Cys Asp Asp Gly Asn Asp Asp Asn Ser 705
710 715 720 Cys Ser Pro Leu Ser Arg Cys Pro Thr Glu Cys Thr Cys Leu
Asp Thr 725 730 735 Val Val Arg Cys Ser Asn Lys Gly Leu Lys Val Leu
Pro Lys Gly Ile 740 745 750 Pro Arg Asp Val Thr Glu Leu Tyr Leu Asp
Gly Asn Gln Phe Thr Leu 755 760 765 Val Pro Lys Glu Leu Ser Asn Tyr
Lys His Leu Thr Leu Ile Asp Leu 770 775 780 Ser Asn Asn Arg Ile Ser
Thr Leu Ser Asn Gln Ser Phe Ser Asn Met 785 790 795 800 Thr Gln Leu
Leu Thr Leu Ile Leu Ser Tyr Asn Arg Leu Arg Cys Ile 805 810 815 Pro
Pro Arg Thr Phe Asp Gly Leu Lys Ser Leu Arg Leu Leu Ser Leu 820 825
830 His Gly Asn Asp Ile Ser Val Val Pro Glu Gly Ala Phe Asn Asp Leu
835 840 845 Ser Ala Leu Ser His Leu Ala Ile Gly Ala Asn Pro Leu Tyr
Cys Asp 850 855 860 Cys Asn Met Gln Trp Leu Ser Asp Trp Val Lys Ser
Glu Tyr Lys Glu 865 870 875 880 Pro Gly Ile Ala Arg Cys Ala Gly Pro
Gly Glu Met Ala Asp Lys Leu 885 890 895 Leu Leu Thr Thr Pro Ser Lys
Lys Phe Thr Cys Gln Gly Pro Val Asp 900 905 910 Val Asn Ile Leu Ala
Lys Cys Asn Pro Cys Leu Ser Asn Pro Cys Lys 915 920 925 Asn Asp Gly
Thr Cys Asn Ser Asp Pro Val Asp Phe Tyr Arg Cys Thr 930 935 940 Cys
Pro Tyr Gly Phe Lys Gly Gln Asp Cys Asp Val Pro Ile His Ala 945 950
955 960 Cys Ile Ser Asn Pro Cys Lys His Gly Gly Thr Cys His Leu Lys
Glu 965 970 975 Gly Glu Glu Asp Gly Phe Trp Cys Ile Cys Ala Asp Gly
Phe Glu Gly 980 985 990 Glu Asn Cys Glu Val Asn Val Asp Asp Cys Glu
Asp Asn Asp Cys Glu 995 1000 1005 Asn Asn Ser Thr Cys Val Asp Gly
Ile Asn Asn Tyr Thr Cys Leu 1010 1015 1020 Cys Pro Pro Glu Tyr Thr
Gly Glu Leu Cys Glu Glu Lys Leu Asp 1025 1030 1035 Phe Cys Ala Gln
Asp Leu Asn Pro Cys Gln His Asp Ser Lys Cys 1040 1045 1050 Ile Leu
Thr Pro Lys Gly Phe Lys Cys Asp Cys Thr Pro Gly Tyr 1055 1060 1065
Val Gly Glu His Cys Asp Ile Asp Phe Asp Asp Cys Gln Asp Asn 1070
1075 1080 Lys Cys Lys Asn Gly Ala His Cys Thr Asp Ala Val Asn Gly
Tyr 1085 1090 1095 Thr Cys Ile Cys Pro Glu Gly Tyr Ser Gly Leu Phe
Cys Glu Phe 1100 1105 1110 Ser Pro Pro Met Val Leu Pro Arg Thr Ser
Pro Cys Asp Asn Phe 1115 1120 1125 Asp Cys Gln Asn Gly Ala Gln Cys
Ile Val Arg Ile Asn Glu Pro 1130 1135 1140 Ile Cys Gln Cys Leu Pro
Gly Tyr Gln Gly Glu Lys Cys Glu Lys 1145 1150 1155 Leu Val Ser Val
Asn Phe Ile Asn Lys Glu Ser Tyr Leu Gln Ile 1160 1165 1170 Pro Ser
Ala Lys Val Arg Pro Gln Thr Asn Ile Thr Leu Gln Ile 1175 1180 1185
Ala Thr Asp Glu Asp Ser Gly Ile Leu Leu Tyr Lys Gly Asp Lys 1190
1195 1200 Asp His Ile Ala Val Glu Leu Tyr Arg Gly Arg Val Arg Ala
Ser 1205 1210 1215 Tyr Asp Thr Gly Ser His Pro Ala Ser Ala Ile Tyr
Ser Val Glu 1220 1225 1230 Thr Ile Asn Asp Gly Asn Phe His Ile Val
Glu Leu Leu Ala Leu 1235 1240 1245 Asp Gln Ser Leu Ser Leu Ser Val
Asp Gly Gly Asn Pro Lys Ile 1250 1255 1260 Ile Thr Asn Leu Ser Lys
Gln Ser Thr Leu Asn Phe Asp Ser Pro 1265 1270 1275 Leu Tyr Val Gly
Gly Met Pro Gly Lys Ser Asn Val Ala Ser Leu 1280 1285 1290 Arg Gln
Ala Pro Gly Gln Asn Gly Thr Ser Phe His Gly Cys Ile 1295 1300 1305
Arg Asn Leu Tyr Ile Asn Ser Glu Leu Gln Asp Phe Gln Lys Val 1310
1315 1320 Pro Met Gln Thr Gly Ile Leu Pro Gly Cys Glu Pro Cys His
Lys 1325 1330 1335 Lys Val Cys Ala His Gly Thr Cys Gln Pro Ser Ser
Gln Ala Gly 1340 1345 1350 Phe Thr Cys Glu Cys Gln Glu Gly Trp Met
Gly Pro Leu Cys Asp 1355 1360 1365 Gln Arg Thr Asn Asp Pro Cys Leu
Gly Asn Lys Cys Val His Gly 1370 1375 1380 Thr Cys Leu Pro Ile Asn
Ala Phe Ser Tyr Ser Cys Lys Cys Leu 1385 1390 1395 Glu Gly His Gly
Gly Val Leu Cys Asp Glu Glu Glu Asp Leu Phe 1400 1405 1410 Asn Pro
Cys Gln Ala Ile Lys Cys Lys His Gly Lys Cys Arg Leu 1415 1420 1425
Ser Gly Leu Gly Gln Pro Tyr Cys Glu Cys Ser Ser Gly Tyr Thr 1430
1435 1440 Gly Asp Ser Cys Asp Arg Glu Ile Ser Cys Arg Gly Glu Arg
Ile 1445 1450 1455 Arg Asp Tyr Tyr Gln Lys Gln Gln Gly Tyr Ala Ala
Cys Gln Thr 1460 1465 1470 Thr Lys Lys Val Ser Arg Leu Glu Cys Arg
Gly Gly Cys Ala Gly 1475 1480 1485 Gly Gln Cys Cys Gly Pro Leu Arg
Ser Lys Arg Arg Lys Tyr Ser 1490 1495 1500 Phe Glu Cys Thr Asp Gly
Ser Ser Phe Val Asp Glu Val Glu Lys 1505 1510 1515 Val Val Lys Cys
Gly Cys Thr Arg Cys Val Ser 1520 1525 <210> SEQ ID NO 30
<211> LENGTH: 4900 <212> TYPE: DNA <213>
ORGANISM: Homo sapiens <400> SEQUENCE: 30 cagagcaggg
tggagagggc ggtgggaggc gtgtgcctga gtgggctcta ctgccttgtt 60
ccatattatt ttgtgcacat tttccctggc actctgggtt gctagccccg ccgggcactg
120 ggcctcagac actgcgcggt tccctcggag cagcaagcta aagaaagccc
ccagtgccgg 180 cgaggaagga ggcggcgggg aaagatgcgc ggcgttggct
ggcagatgct gtccctgtcg 240 ctggggttag tgctggcgat cctgaacaag
gtggcaccgc aggcgtgccc ggcgcagtgc 300 tcttgctcgg gcagcacagt
ggactgtcac gggctggcgc tgcgcagcgt gcccaggaat 360 atcccccgca
acaccgagag actggattta aatggaaata acatcacaag aattacgaag 420
acagattttg ctggtcttag acatctaaga gttcttcagc ttatggagaa taagattagc
480 accattgaaa gaggagcatt ccaggatctt aaagaactag agagactgcg
tttaaacaga 540 aatcaccttc agctgtttcc tgagttgctg tttcttggga
ctgcgaagct atacaggctt 600 gatctcagtg aaaaccaaat tcaggcaatc
ccaaggaaag ctttccgtgg ggcagttgac 660 ataaaaaatt tgcaactgga
ttacaaccag atcagctgta ttgaagatgg ggcattcagg 720 gctctccggg
acctggaagt gctcactctc aacaataaca acattactag actttctgtg 780
gcaagtttca accatatgcc taaacttagg acttttcgac tgcattcaaa caacctgtat
840 tgtgactgcc acctggcctg gctctccgac tggcttcgcc aaaggcctcg
ggttggtctg 900 tacactcagt gtatgggccc ctcccacctg agaggccata
atgtagccga ggttcaaaaa 960 cgagaatttg tctgcagtgg tcaccagtca
tttatggctc cttcttgtag tgttttgcac 1020 tgccctgccg cctgtacctg
tagcaacaat atcgtagact gtcgtgggaa aggtctcact 1080 gagatcccca
caaatcttcc agagaccatc acagaaatac gtttggaaca gaacacaatc 1140
aaagtcatcc ctcctggagc tttctcacca tataaaaagc ttagacgaat tgacctgagc
1200 aataatcaga tctctgaact tgcaccagat gctttccaag gactacgctc
tctgaattca 1260 cttgtcctct atggaaataa aatcacagaa ctccccaaaa
gtttatttga aggactgttt 1320 tccttacagc tcctattatt gaatgccaac
aagataaact gccttcgggt agatgctttt 1380 caggatctcc acaacttgaa
ccttctctcc ctatatgaca acaagcttca gaccatcgcc 1440 aaggggacct
tttcacctct tcgggccatt caaactatgc atttggccca gaaccccttt 1500
atttgtgact gccatctcaa gtggctagcg gattatctcc ataccaaccc gattgagacc
1560 agtggtgccc gttgcaccag cccccgccgc ctggcaaaca aaagaattgg
acagatcaaa 1620 agcaagaaat tccgttgttc agctaaagaa cagtatttca
ttccaggtac agaagattat 1680 cgatcaaaat taagtggaga ctgctttgcg
gatctggctt gccctgaaaa gtgtcgctgt 1740 gaaggaacca cagtagattg
ctctaatcaa aagctcaaca aaatcccgga gcacattccc 1800 cagtacactg
cagagttgcg tctcaataat aatgaattta ccgtgttgga agccacagga 1860
atctttaaga aacttcctca attacgtaaa ataaacttta gcaacaataa gatcacagat
1920 attgaggagg gagcatttga aggagcatct ggtgtaaatg aaatacttct
tacgagtaat 1980 cgtttggaaa atgtgcagca taagatgttc aagggattgg
aaagcctcaa aactttgatg 2040 ttgagaagca atcgaataac ctgtgtgggg
aatgacagtt tcataggact cagttctgtg 2100 cgtttgcttt ctttgtatga
taatcaaatt actacagttg caccaggggc atttgatact 2160 ctccattctt
tatctactct aaacctcttg gccaatcctt ttaactgtaa ctgctacctg 2220
gcttggttgg gagagtggct gagaaagaag agaattgtca cgggaaatcc tagatgtcaa
2280 aaaccatact tcctgaaaga aatacccatc caggatgtgg ccattcagga
cttcacttgt 2340 gatgacggaa atgatgacaa tagttgctcc ccactttctc
gctgtcctac tgaatgtact 2400 tgcttggata cagtcgtccg atgtagcaac
aagggtttga aggtcttgcc gaaaggtatt 2460 ccaagagatg tcacagagtt
gtatctggat ggaaaccaat ttacactggt tcccaaggaa 2520 ctctccaact
acaaacattt aacacttata gacttaagta acaacagaat aagcacgctt 2580
tctaatcaga gcttcagcaa catgacccag ctcctcacct taattcttag ttacaaccgt
2640 ctgagatgta ttcctcctcg cacctttgat ggattaaagt ctcttcgatt
actttctcta 2700 catggaaatg acatttctgt tgtgcctgaa ggtgctttca
atgatctttc tgcattatca 2760 catctagcaa ttggagccaa ccctctttac
tgtgattgta acatgcagtg gttatccgac 2820 tgggtgaagt cggaatataa
ggagcctgga attgctcgtt gtgctggtcc tggagaaatg 2880 gcagataaac
ttttactcac aactccctcc aaaaaattta cctgtcaagg tcctgtggat 2940
gtcaatattc tagctaagtg taacccctgc ctatcaaatc cgtgtaaaaa tgatggcaca
3000 tgtaatagtg atccagttga cttttaccga tgcacctgtc catatggttt
caaggggcag 3060 gactgtgatg tcccaattca tgcctgcatc agtaacccat
gtaaacatgg aggaacttgc 3120 cacttaaagg aaggagaaga agatggattc
tggtgtattt gtgctgatgg atttgaagga 3180 gaaaattgtg aagtcaacgt
tgatgattgt gaagataatg actgtgaaaa taattctaca 3240 tgtgtcgatg
gcattaataa ctacacatgc ctttgcccac ctgagtatac aggtgagttg 3300
tgtgaggaga agctggactt ctgtgcccag gacctgaacc cctgccagca cgattcaaag
3360 tgcatcctaa ctccaaaggg attcaaatgt gactgcacac cagggtacgt
aggtgaacac 3420 tgcgacatcg attttgacga ctgccaagac aacaagtgta
aaaacggagc ccactgcaca 3480 gatgcagtga acggctatac gtgcatatgc
cccgaaggtt acagtggctt gttctgtgag 3540 ttttctccac ccatggtcct
ccctcgtacc agcccctgtg ataattttga ttgtcagaat 3600 ggagctcagt
gtatcgtcag aataaatgag ccaatatgtc agtgtttgcc tggctatcag 3660
ggagaaaagt gtgaaaaatt ggttagtgtg aattttataa acaaagagtc ttatcttcag
3720 attccttcag ccaaggttcg gcctcagacg aacataacac ttcagattgc
cacagatgaa 3780 gacagcggaa tcctcctgta taagggtgac aaagaccata
tcgcggtaga actctatcgg 3840 gggcgtgttc gtgccagcta tgacaccggc
tctcatccag cttctgccat ttacagtgtg 3900 gagacaatca atgatggaaa
cttccacatt gtggaactac ttgccttgga tcagagtctc 3960 tctttgtccg
tggatggtgg gaaccccaaa atcatcacta acttgtcaaa gcagtccact 4020
ctgaattttg actctccact ctatgtagga ggcatgccag ggaagagtaa cgtggcatct
4080 ctgcgccagg cccctgggca gaacggaacc agcttccacg gctgcatccg
gaacctttac 4140 atcaacagtg agctgcagga cttccagaag gtgccgatgc
aaacaggcat tttgcctggc 4200 tgtgagccat gccacaagaa ggtgtgtgcc
catggcacat gccagcccag cagccaggca 4260 ggcttcacct gcgagtgcca
ggaaggatgg atggggcccc tctgtgacca acggaccaat 4320 gacccttgcc
ttggaaataa atgcgtacat ggcacctgct tgcccatcaa tgcgttctcc 4380
tacagctgta agtgcttgga gggccatgga ggtgtcctct gtgatgaaga ggaggatctg
4440 tttaacccat gccaggcgat caagtgcaag cacgggaagt gcaggctttc
aggtctgggg 4500 cagccctact gtgaatgcag cagtggatac acgggggaca
gctgtgatcg agaaatctct 4560 tgtcgagggg aaaggataag agattattac
caaaagcagc agggctatgc tgcttgccaa 4620 acaaccaaga aggtgtcccg
attagagtgc agaggtgggt gtgcaggagg gcagtgctgt 4680 ggaccgctga
ggagcaagcg gcggaaatac tctttcgaat gcactgacgg ctcctccttt 4740
gtggacgagg ttgagaaagt ggtgaagtgc ggctgtacga ggtgtgtgtc ctaaacacac
4800 tcccggcagc tctgtctttg gaaaaggttg tatacttctt gaccatgtgg
gactaatgaa 4860 tgcttcatag tggaaatatt tgaaatatat tgtaaaatac 4900
<210> SEQ ID NO 31 <211> LENGTH: 3510 <212> TYPE:
DNA <213> ORGANISM: Homo sapiens <400> SEQUENCE: 31
gcagctctgg gggagctcgg agctcccgat cacggcttct tgggggtagc tacggctggg
60 tgtgtagaac ggggccgggg ctggggctgg gtcccctagt ggagacccaa
gtgcgagagg 120 caagaactct gcagcttcct gccttctggg tcagttcctt
attcaagtct gcagccggct 180 cccagggaga tctcggtgga acttcagaaa
cgctgggcag tctgcctttc aaccatgccc 240 ctgtccctgg gagccgagat
gtgggggcct gaggcctggc tgctgctgct gctactgctg 300 gcatcattta
caggccggtg ccccgcgggt gagctggaga cctcagacgt ggtaactgtg 360
gtgctgggcc aggacgcaaa actgccctgc ttctaccgag gggactccgg cgagcaagtg
420 gggcaagtgg catgggctcg ggtggacgcg ggcgaaggcg cccaggaact
agcgctactg 480 cactccaaat acgggcttca tgtgagcccg gcttacgagg
gccgcgtgga gcagccgccg 540 cccccacgca accccctgga cggctcagtg
ctcctgcgca acgcagtgca ggcggatgag 600 ggcgagtacg agtgccgggt
cagcaccttc cccgccggca gcttccaggc gcggctgcgg 660 ctccgagtgc
tggtgcctcc cctgccctca ctgaatcctg gtccagcact agaagagggc 720
cagggcctga ccctggcagc ctcctgcaca gctgagggca gcccagcccc cagcgtgacc
780 tgggacacgg aggtcaaagg cacaacgtcc agccgttcct tcaagcactc
ccgctctgct 840 gccgtcacct cagagttcca cttggtgcct agccgcagca
tgaatgggca gccactgact 900 tgtgtggtgt cccatcctgg cctgctccag
gaccaaagga tcacccacat cctccacgtg 960 tccttccttg ctgaggcctc
tgtgaggggc cttgaagacc aaaatctgtg gcacattggc 1020 agagaaggag
ctatgctcaa gtgcctgagt gaagggcagc cccctccctc atacaactgg 1080
acacggctgg atgggcctct gcccagtggg gtacgagtgg atggggacac tttgggcttt
1140 cccccactga ccactgagca cagcggcatc tacgtctgcc atgtcagcaa
tgagttctcc 1200 tcaagggatt ctcaggtcac tgtggatgtt cttgaccccc
aggaagactc tgggaagcag 1260 gtggacctag tgtcagcctc ggtggtggtg
gtgggtgtga tcgccgcact cttgttctgc 1320 cttctggtgg tggtggtggt
gctcatgtcc cgataccatc ggcgcaaggc ccagcagatg 1380 acccagaaat
atgaggagga gctgaccctg accagggaga actccatccg gaggctgcat 1440
tcccatcaca cggaccccag gagccagccg gaggagagtg tagggctgag agccgagggc
1500 caccctgata gtctcaagga caacagtagc tgctctgtga tgagtgaaga
gcccgagggc 1560 cgcagttact ccacgctgac cacggtgagg gagatagaaa
cacagactga actgctgtct 1620 ccaggctctg ggcgggccga ggaggaggaa
gatcaggatg aaggcatcaa acaggccatg 1680 aaccattttg ttcaggagaa
tgggacccta cgggccaagc ccacgggcaa tggcatctac 1740 atcaatgggc
ggggacacct ggtctgaccc aggcctgcct cccttcccta ggcctggctc 1800
cttctgttga catgggagat tttagctcat cttgggggcc tccttaaaca cccccatttc
1860 ttgcggaaga tgctccccat cccactgact gcttgacctt tacctccaac
ccttctgttc 1920 atcgggaggg ctccaccaat tgagtctctc ccaccatgca
tgcaggtcac tgtgtgtgtg 1980 catgtgtgcc tgtgtgagtg ttgactgact
gtgtgtgtgt ggaggggtga ctgtccgtgg 2040 aggggtgact gtgtccgtgg
tgtgtattat gctgtcatat cagagtcaag tgaactgtgg 2100 tgtatgtgcc
acgggatttg agtggttgcg tgggcaacac tgtcagggtt tggcgtgtgt 2160
gtcatgtggc tgtgtgtgac ctctgcctga aaaagcaggt attttctcag accccagagc
2220 agtattaatg atgcagaggt tggaggagag aggtggagac tgtggctcag
acccaggtgt 2280 gcgggcatag ctggagctgg aatctgcctc cggtgtgagg
gaacctgtct cctaccactt 2340 cggagccatg ggggcaagtg tgaagcagcc
agtccctggg tcagccagag gcttgaactg 2400 ttacagaagc cctctgccct
ctggtggcct ctgggcctgc tgcatgtaca tattttctgt 2460 aaatatacat
gcgccgggag cttcttgcag gaatactgct ccgaatcact tttaattttt 2520
ttcttttttt tttcttgccc tttccattag ttgtattttt tatttatttt tatttttatt
2580 tttttttaga gatggagtct cactatgttg ctcaggctgg ccttgaactc
ctgggctcaa 2640 gcaatcctcc tgcctcagcc tccctagtag ctgggacttt
aagtgtacac cactgtgcct 2700 gctttgaatc ctttacgaag agaaaaaaaa
aattaaagaa agcctttaga tttatccaat 2760 gtttactact gggattgctt
aaagtgaggc ccctccaaca ccagggggtt aattcctgtg 2820 attgtgaaag
gggctacttc caaggcatct tcatgcaggc agccccttgg gagggcacct 2880
gagagctggt agagtctgaa attagggatg tgagcctcgt ggttactgag taaggtaaaa
2940 ttgcatccac cattgtttgt gataccttag ggaattgctt ggacctggtg
acaagggctc 3000 ctgttcaata gtggtgttgg ggagagagag agcagtgatt
atagaccgag agagtaggag 3060 ttgaggtgag gtgaaggagg tgctgggggt
gagaatgtcg cctttccccc tgggttttgg 3120 atcactaatt caaggctctt
ctggatgttt ctctgggttg gggctggagt tcaatgaggt 3180 ttatttttag
ctggcccacc cagatacact cagccagaat acctagattt agtacccaaa 3240
ctcttcttag tctgaaatct gctggatttc tggcctaagg gagaggctcc catccttcgt
3300 tccccagcca gcctaggact tcgaatgtgg agcctgaaga tctaagatcc
taacatgtac 3360 attttatgta aatatgtgca tatttgtaca taaaatgata
ttctgttttt aaataaacag 3420 acaaaacttg aaaaaaaaaa aaaaaaaaaa
aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa 3480 aaaaaaaaaa aaaaaaaaaa
aaaaaaaaaa 3510 <210> SEQ ID NO 32 <211> LENGTH: 1530
<212> TYPE: DNA <213> ORGANISM: Homo sapiens
<400> SEQUENCE: 32 atgcccctgt ccctgggagc cgagatgtgg
gggcctgagg cctggctgct gctgctgcta 60 ctgctggcat catttacagg
ccggtgcccc gcgggtgagc tggagacctc agacgtggta 120 actgtggtgc
tgggccagga cgcaaaactg ccctgcttct accgagggga ctccggcgag 180
caagtggggc aagtggcatg ggctcgggtg gacgcgggcg aaggcgccca ggaactagcg
240 ctactgcact ccaaatacgg gcttcatgtg agcccggctt acgagggccg
cgtggagcag 300 ccgccgcccc cacgcaaccc cctggacggc tcagtgctcc
tgcgcaacgc agtgcaggcg 360 gatgagggcg agtacgagtg ccgggtcagc
accttccccg ccggcagctt ccaggcgcgg 420 ctgcggctcc gagtgctggt
gcctcccctg ccctcactga atcctggtcc agcactagaa 480 gagggccagg
gcctgaccct ggcagcctcc tgcacagctg agggcagccc agcccccagc 540
gtgacctggg acacggaggt caaaggcaca acgtccagcc gttccttcaa gcactcccgc
600 tctgctgccg tcacctcaga gttccacttg gtgcctagcc gcagcatgaa
tgggcagcca 660 ctgacttgtg tggtgtccca tcctggcctg ctccaggacc
aaaggatcac ccacatcctc 720 cacgtgtcct tccttgctga ggcctctgtg
aggggccttg aagaccaaaa tctgtggcac 780 attggcagag aaggagctat
gctcaagtgc ctgagtgaag ggcagccccc tccctcatac 840 aactggacac
ggctggatgg gcctctgccc agtggggtac gagtggatgg ggacactttg 900
ggctttcccc cactgaccac tgagcacagc ggcatctacg tctgccatgt cagcaatgag
960 ttctcctcaa gggattctca ggtcactgtg gatgttcttg acccccagga
agactctggg 1020 aagcaggtgg acctagtgtc agcctcggtg gtggtggtgg
gtgtgatcgc cgcactcttg 1080 ttctgccttc tggtggtggt ggtggtgctc
atgtcccgat accatcggcg caaggcccag 1140 cagatgaccc agaaatatga
ggaggagctg accctgacca gggagaactc catccggagg 1200 ctgcattccc
atcacacgga ccccaggagc cagccggagg agagtgtagg gctgagagcc 1260
gagggccacc ctgatagtct caaggacaac agtagctgct ctgtgatgag tgaagagccc
1320 gagggccgca gttactccac gctgaccacg gtgagggaga tagaaacaca
gactgaactg 1380 ctgtctccag gctctgggcg ggccgaggag gaggaagatc
aggatgaagg catcaaacag 1440 gccatgaacc attttgttca ggagaatggg
accctacggg ccaagcccac gggcaatggc 1500 atctacatca atgggcgggg
acacctggtc 1530 <210> SEQ ID NO 33 <211> LENGTH: 510
<212> TYPE: PRT <213> ORGANISM: Homo sapiens
<400> SEQUENCE: 33 Met Pro Leu Ser Leu Gly Ala Glu Met Trp
Gly Pro Glu Ala Trp Leu 1 5 10 15 Leu Leu Leu Leu Leu Leu Ala Ser
Phe Thr Gly Arg Cys Pro Ala Gly 20 25 30 Glu Leu Glu Thr Ser Asp
Val Val Thr Val Val Leu Gly Gln Asp Ala 35 40 45 Lys Leu Pro Cys
Phe Tyr Arg Gly Asp Ser Gly Glu Gln Val Gly Gln 50 55 60 Val Ala
Trp Ala Arg Val Asp Ala Gly Glu Gly Ala Gln Glu Leu Ala 65 70 75 80
Leu Leu His Ser Lys Tyr Gly Leu His Val Ser Pro Ala Tyr Glu Gly 85
90 95 Arg Val Glu Gln Pro Pro Pro Pro Arg Asn Pro Leu Asp Gly Ser
Val 100 105 110 Leu Leu Arg Asn Ala Val Gln Ala Asp Glu Gly Glu Tyr
Glu Cys Arg 115 120 125 Val Ser Thr Phe Pro Ala Gly Ser Phe Gln Ala
Arg Leu Arg Leu Arg 130 135 140 Val Leu Val Pro Pro Leu Pro Ser Leu
Asn Pro Gly Pro Ala Leu Glu 145 150 155 160 Glu Gly Gln Gly Leu Thr
Leu Ala Ala Ser Cys Thr Ala Glu Gly Ser 165 170 175 Pro Ala Pro Ser
Val Thr Trp Asp Thr Glu Val Lys Gly Thr Thr Ser 180 185 190 Ser Arg
Ser Phe Lys His Ser Arg Ser Ala Ala Val Thr Ser Glu Phe 195 200 205
His Leu Val Pro Ser Arg Ser Met Asn Gly Gln Pro Leu Thr Cys Val 210
215 220 Val Ser His Pro Gly Leu Leu Gln Asp Gln Arg Ile Thr His Ile
Leu 225 230 235 240 His Val Ser Phe Leu Ala Glu Ala Ser Val Arg Gly
Leu Glu Asp Gln 245 250 255 Asn Leu Trp His Ile Gly Arg Glu Gly Ala
Met Leu Lys Cys Leu Ser 260 265 270 Glu Gly Gln Pro Pro Pro Ser Tyr
Asn Trp Thr Arg Leu Asp Gly Pro 275 280 285 Leu Pro Ser Gly Val Arg
Val Asp Gly Asp Thr Leu Gly Phe Pro Pro 290 295 300 Leu Thr Thr Glu
His Ser Gly Ile Tyr Val Cys His Val Ser Asn Glu 305 310 315 320 Phe
Ser Ser Arg Asp Ser Gln Val Thr Val Asp Val Leu Asp Pro Gln 325 330
335 Glu Asp Ser Gly Lys Gln Val Asp Leu Val Ser Ala Ser Val Val Val
340 345 350 Val Gly Val Ile Ala Ala Leu Leu Phe Cys Leu Leu Val Val
Val Val 355 360 365 Val Leu Met Ser Arg Tyr His Arg Arg Lys Ala Gln
Gln Met Thr Gln 370 375 380 Lys Tyr Glu Glu Glu Leu Thr Leu Thr Arg
Glu Asn Ser Ile Arg Arg 385 390 395 400 Leu His Ser His His Thr Asp
Pro Arg Ser Gln Pro Glu Glu Ser Val 405 410 415 Gly Leu Arg Ala Glu
Gly His Pro Asp Ser Leu Lys Asp Asn Ser Ser 420 425 430 Cys Ser Val
Met Ser Glu Glu Pro Glu Gly Arg Ser Tyr Ser Thr Leu 435 440 445 Thr
Thr Val Arg Glu Ile Glu Thr Gln Thr Glu Leu Leu Ser Pro Gly 450 455
460 Ser Gly Arg Ala Glu Glu Glu Glu Asp Gln Asp Glu Gly Ile Lys Gln
465 470 475 480 Ala Met Asn His Phe Val Gln Glu Asn Gly Thr Leu Arg
Ala Lys Pro 485 490 495 Thr Gly Asn Gly Ile Tyr Ile Asn Gly Arg Gly
His Leu Val 500 505 510 <210> SEQ ID NO 34 <211>
LENGTH: 31 <212> TYPE: PRT <213> ORGANISM: Homo sapiens
<400> SEQUENCE: 34 Met Pro Leu Ser Leu Gly Ala Glu Met Trp
Gly Pro Glu Ala Trp Leu 1 5 10 15 Leu Leu Leu Leu Leu Leu Ala Ser
Phe Thr Gly Arg Cys Pro Ala 20 25 30 <210> SEQ ID NO 35
<211> LENGTH: 479 <212> TYPE: PRT <213> ORGANISM:
Homo sapiens <400> SEQUENCE: 35 Gly Glu Leu Glu Thr Ser Asp
Val Val Thr Val Val Leu Gly Gln Asp 1 5 10 15 Ala Lys Leu Pro Cys
Phe Tyr Arg Gly Asp Ser Gly Glu Gln Val Gly 20 25 30 Gln Val Ala
Trp Ala Arg Val Asp Ala Gly Glu Gly Ala Gln Glu Leu 35 40 45 Ala
Leu Leu His Ser Lys Tyr Gly Leu His Val Ser Pro Ala Tyr Glu 50 55
60 Gly Arg Val Glu Gln Pro Pro Pro Pro Arg Asn Pro Leu Asp Gly Ser
65 70 75 80 Val Leu Leu Arg Asn Ala Val Gln Ala Asp Glu Gly Glu Tyr
Glu Cys 85 90 95 Arg Val Ser Thr Phe Pro Ala Gly Ser Phe Gln Ala
Arg Leu Arg Leu 100 105 110 Arg Val Leu Val Pro Pro Leu Pro Ser Leu
Asn Pro Gly Pro Ala Leu 115 120 125 Glu Glu Gly Gln Gly Leu Thr Leu
Ala Ala Ser Cys Thr Ala Glu Gly 130 135 140 Ser Pro Ala Pro Ser Val
Thr Trp Asp Thr Glu Val Lys Gly Thr Thr 145 150 155 160 Ser Ser Arg
Ser Phe Lys His Ser Arg Ser Ala Ala Val Thr Ser Glu 165 170 175 Phe
His Leu Val Pro Ser Arg Ser Met Asn Gly Gln Pro Leu Thr Cys 180 185
190 Val Val Ser His Pro Gly Leu Leu Gln Asp Gln Arg Ile Thr His Ile
195 200 205 Leu His Val Ser Phe Leu Ala Glu Ala Ser Val Arg Gly Leu
Glu Asp 210 215 220 Gln Asn Leu Trp His Ile Gly Arg Glu Gly Ala Met
Leu Lys Cys Leu 225 230 235 240 Ser Glu Gly Gln Pro Pro Pro Ser Tyr
Asn Trp Thr Arg Leu Asp Gly 245 250 255 Pro Leu Pro Ser Gly Val Arg
Val Asp Gly Asp Thr Leu Gly Phe Pro 260 265 270 Pro Leu Thr Thr Glu
His Ser Gly Ile Tyr Val Cys His Val Ser Asn 275 280 285 Glu Phe Ser
Ser Arg Asp Ser Gln Val Thr Val Asp Val Leu Asp Pro 290 295 300 Gln
Glu Asp Ser Gly Lys Gln Val Asp Leu Val Ser Ala Ser Val Val 305 310
315 320 Val Val Gly Val Ile Ala Ala Leu Leu Phe Cys Leu Leu Val Val
Val 325 330 335 Val Val Leu Met Ser Arg Tyr His Arg Arg Lys Ala Gln
Gln Met Thr 340 345 350 Gln Lys Tyr Glu Glu Glu Leu Thr Leu Thr Arg
Glu Asn Ser Ile Arg 355 360 365 Arg Leu His Ser His His Thr Asp Pro
Arg Ser Gln Pro Glu Glu Ser 370 375 380 Val Gly Leu Arg Ala Glu Gly
His Pro Asp Ser Leu Lys Asp Asn Ser 385 390 395 400 Ser Cys Ser Val
Met Ser Glu Glu Pro Glu Gly Arg Ser Tyr Ser Thr 405 410 415 Leu Thr
Thr Val Arg Glu Ile Glu Thr Gln Thr Glu Leu Leu Ser Pro 420 425 430
Gly Ser Gly Arg Ala Glu Glu Glu Glu Asp Gln Asp Glu Gly Ile Lys 435
440 445 Gln Ala Met Asn His Phe Val Gln Glu Asn Gly Thr Leu Arg Ala
Lys 450 455 460 Pro Thr Gly Asn Gly Ile Tyr Ile Asn Gly Arg Gly His
Leu Val 465 470 475 <210> SEQ ID NO 36 <211> LENGTH:
314 <212> TYPE: PRT <213> ORGANISM: Homo sapiens
<400> SEQUENCE: 36 Gly Glu Leu Glu Thr Ser Asp Val Val Thr
Val Val Leu Gly Gln Asp 1 5 10 15 Ala Lys Leu Pro Cys Phe Tyr Arg
Gly Asp Ser Gly Glu Gln Val Gly 20 25 30 Gln Val Ala Trp Ala Arg
Val Asp Ala Gly Glu Gly Ala Gln Glu Leu 35 40 45 Ala Leu Leu His
Ser Lys Tyr Gly Leu His Val Ser Pro Ala Tyr Glu 50 55 60 Gly Arg
Val Glu Gln Pro Pro Pro Pro Arg Asn Pro Leu Asp Gly Ser 65 70 75 80
Val Leu Leu Arg Asn Ala Val Gln Ala Asp Glu Gly Glu Tyr Glu Cys 85
90 95 Arg Val Ser Thr Phe Pro Ala Gly Ser Phe Gln Ala Arg Leu Arg
Leu 100 105 110 Arg Val Leu Val Pro Pro Leu Pro Ser Leu Asn Pro Gly
Pro Ala Leu 115 120 125 Glu Glu Gly Gln Gly Leu Thr Leu Ala Ala Ser
Cys Thr Ala Glu Gly 130 135 140 Ser Pro Ala Pro Ser Val Thr Trp Asp
Thr Glu Val Lys Gly Thr Thr 145 150 155 160 Ser Ser Arg Ser Phe Lys
His Ser Arg Ser Ala Ala Val Thr Ser Glu 165 170 175 Phe His Leu Val
Pro Ser Arg Ser Met Asn Gly Gln Pro Leu Thr Cys 180 185 190 Val Val
Ser His Pro Gly Leu Leu Gln Asp Gln Arg Ile Thr His Ile 195 200 205
Leu His Val Ser Phe Leu Ala Glu Ala Ser Val Arg Gly Leu Glu Asp 210
215 220 Gln Asn Leu Trp His Ile Gly Arg Glu Gly Ala Met Leu Lys Cys
Leu 225 230 235 240 Ser Glu Gly Gln Pro Pro Pro Ser Tyr Asn Trp Thr
Arg Leu Asp Gly 245 250 255 Pro Leu Pro Ser Gly Val Arg Val Asp Gly
Asp Thr Leu Gly Phe Pro 260 265 270 Pro Leu Thr Thr Glu His Ser Gly
Ile Tyr Val Cys His Val Ser Asn 275 280 285 Glu Phe Ser Ser Arg Asp
Ser Gln Val Thr Val Asp Val Leu Asp Pro 290 295 300 Gln Glu Asp Ser
Gly Lys Gln Val Asp Leu 305 310 <210> SEQ ID NO 37
<211> LENGTH: 25 <212> TYPE: PRT <213> ORGANISM:
Homo sapiens <400> SEQUENCE: 37 Val Ser Ala Ser Val Val Val
Val Gly Val Ile Ala Ala Leu Leu Phe 1 5 10 15 Cys Leu Leu Val Val
Val Val Val Leu 20 25 <210> SEQ ID NO 38 <211> LENGTH:
140 <212> TYPE: PRT <213> ORGANISM: Homo sapiens
<400> SEQUENCE: 38 Met Ser Arg Tyr His Arg Arg Lys Ala Gln
Gln Met Thr Gln Lys Tyr 1 5 10 15 Glu Glu Glu Leu Thr Leu Thr Arg
Glu Asn Ser Ile Arg Arg Leu His 20 25 30 Ser His His Thr Asp Pro
Arg Ser Gln Pro Glu Glu Ser Val Gly Leu 35 40 45 Arg Ala Glu Gly
His Pro Asp Ser Leu Lys Asp Asn Ser Ser Cys Ser 50 55 60 Val Met
Ser Glu Glu Pro Glu Gly Arg Ser Tyr Ser Thr Leu Thr Thr 65 70 75 80
Val Arg Glu Ile Glu Thr Gln Thr Glu Leu Leu Ser Pro Gly Ser Gly 85
90 95 Arg Ala Glu Glu Glu Glu Asp Gln Asp Glu Gly Ile Lys Gln Ala
Met 100 105 110 Asn His Phe Val Gln Glu Asn Gly Thr Leu Arg Ala Lys
Pro Thr Gly 115 120 125 Asn Gly Ile Tyr Ile Asn Gly Arg Gly His Leu
Val 130 135 140 <210> SEQ ID NO 39 <400> SEQUENCE: 39
000 3 <210> SEQ ID NO 40 <400> SEQUENCE: 40 000 3
<210> SEQ ID NO 41 <211> LENGTH: 2510 <212> TYPE:
DNA <213> ORGANISM: Homo sapiens <400> SEQUENCE: 41
caaaggcaca acgtccagcc gttccttcaa gcactcccgc tctgctgccg tcacctcaga
60 gttccacttg gtgcctagcc gcagcatgaa tgggcagcca ctgacttgtg
tggtgtccca 120 tcctggcctg ctccaggacc aaaggatcac ccacatcctc
cacgtgtcct tccttgctga 180 ggcctctgtg aggggccttg aagaccaaaa
tctgtggcac attggcagag aaggagctat 240 gctcaagtgc ctgagtgaag
ggcagccccc tccctcatac aactggacac ggctggatgg 300 gcctctgccc
agtggggtac gagtggatgg ggacactttg ggctttcccc cactgaccac 360
tgagcacagc ggcatctacg tctgccatgt cagcaatgag ttctcctcaa gggattctca
420 ggtcactgtg gatgttcttg cagaccccca ggaagactct gggaagcagg
tggacctagt 480 gtcagcctcg gtggtggtgg tgggtgtgat cgccgcactc
ttgttctgcc ttctggtggt 540 ggtggtggtg ctcatgtccc gataccatcg
gcgcaaggcc cagcagatga cccagaaata 600 tgaggaggag ctgaccctga
ccagggagaa ctccatccgg aggctgcatt cccatcacac 660 ggaccccagg
agccagagtg aagagcccga gggccgcagt tactccacgc tgaccacggt 720
gagggagata gaaacacaga ctgaactgct gtctccaggc tctgggcggg ccgaggagga
780 ggaagatcag gatgaaggca tcaaacaggc catgaaccat tttgttcagg
agaatgggac 840 cctacgggcc aagcccacgg gcaatggcat ctacatcaat
gggcggggac acctggtctg 900 acccaggcct gcctcccttc cctaggcctg
gctccttctg ttgacatggg agattttagc 960 tcatcttggg ggcctcctta
aacaccccca tttcttgcgg aagatgctcc ccatcccact 1020 gactgcttga
cctttacctc caacccttct gttcatcggg agggctccac caattgagtc 1080
tctcccacca tgcatgcagg tcactgtgtg tgtgcatgtg tgcctgtgtg agtgttgact
1140 gactgtgtgt gtgtggaggg gtgactgtcc gtggaggggt gactgtgtcc
gtggtgtgta 1200 ttatgctgtc atatcagagt caagtgaact gtggtgtatg
tgccacggga tttgagtggt 1260 tgcgtgggca acactgtcag ggtttggcgt
gtgtgtcatg tggctgtgtg tgacctctgc 1320 ctgaaaaagc aggtattttc
tcagacccca gagcagtatt aatgatgcag aggttggagg 1380 agagaggtgg
agactgtggc tcagacccag gtgtgcgggc atagctggag ctggaatctg 1440
cctccggtgt gagggaacct gtctcctacc acttcggagc catgggggca agtgtgaagc
1500 agccagtccc tgggtcagcc agaggcttga actgttacag aagccctctg
ccctctggtg 1560 gcctctgggc ctgctgcatg tacatatttt ctgtaaatat
acatgcgccg ggagcttctt 1620 gcaggaatac tgctccgaat cacttttaat
ttttttcttt tttttttctt gccctttcca 1680 ttagttgtat tttttattta
tttttatttt tatttttttt tagagatgga gtctcactat 1740 gttgctcagg
ctggccttga actcctgggc tcaagcaatc ctcctgcctc agcctcccta 1800
gtagctggga ctttaagtgt acaccactgt gcctgctttg aatcctttac gaagagaaaa
1860 aaaaaattaa agaaagcctt tagatttatc caatgtttac tactgggatt
gcttaaagtg 1920 aggcccctcc aacaccaggg ggttaattcc tgtgattgtg
aaaggggcta cttccaaggc 1980 atcttcatgc aggcagcccc ttgggagggc
acctgagagc tggtagagtc tgaaattagg 2040 gatgtgagcc tggtgacaag
ggctcctgtt caatagtggt gttggggaga gagagagcag 2100 tgattataga
ccgagagagt aggagttgag gtgaggtgaa ggaggtgctg ggggtgagaa 2160
tgtcgccttt ccccctgggt tttggatcac taattcaagg ctcttctgga tgtttctctg
2220 ggttggggct ggagttcaat gaggtttatt tttagctggc ccacccagat
acactcagcc 2280 agaataccta gatttagtac ccaaactctt cttagtctga
aatctgctgg atttctggcc 2340 taagggagag gctcccatcc ttcgttcccc
agccagccta ggacttcgaa tgtggagcct 2400 gaagatctaa gatcctaaca
tgtacatttt atgtaaatat gtgcatattt gtacataaaa 2460 tgatattctg
tttttaaata aacagacaaa acttgaaaaa aaaaaaaaaa 2510 <210> SEQ ID
NO 42 <211> LENGTH: 897 <212> TYPE: DNA <213>
ORGANISM: Homo sapiens <400> SEQUENCE: 42 aaaggcacaa
cgtccagccg ttccttcaag cactcccgct ctgctgccgt cacctcagag 60
ttccacttgg tgcctagccg cagcatgaat gggcagccac tgacttgtgt ggtgtcccat
120 cctggcctgc tccaggacca aaggatcacc cacatcctcc acgtgtcctt
ccttgctgag 180 gcctctgtga ggggccttga agaccaaaat ctgtggcaca
ttggcagaga aggagctatg 240 ctcaagtgcc tgagtgaagg gcagccccct
ccctcataca actggacacg gctggatggg 300 cctctgccca gtggggtacg
agtggatggg gacactttgg gctttccccc actgaccact 360 gagcacagcg
gcatctacgt ctgccatgtc agcaatgagt tctcctcaag ggattctcag 420
gtcactgtgg atgttcttgc agacccccag gaagactctg ggaagcaggt ggacctagtg
480 tcagcctcgg tggtggtggt gggtgtgatc gccgcactct tgttctgcct
tctggtggtg 540 gtggtggtgc tcatgtcccg ataccatcgg cgcaaggccc
agcagatgac ccagaaatat 600 gaggaggagc tgaccctgac cagggagaac
tccatccgga ggctgcattc ccatcacacg 660 gaccccagga gccagagtga
agagcccgag ggccgcagtt actccacgct gaccacggtg 720 agggagatag
aaacacagac tgaactgctg tctccaggct ctgggcgggc cgaggaggag 780
gaagatcagg atgaaggcat caaacaggcc atgaaccatt ttgttcagga gaatgggacc
840 ctacgggcca agcccacggg caatggcatc tacatcaatg ggcggggaca cctggtc
897 <210> SEQ ID NO 43 <211> LENGTH: 299 <212>
TYPE: PRT <213> ORGANISM: Homo sapiens <400> SEQUENCE:
43 Lys Gly Thr Thr Ser Ser Arg Ser Phe Lys His Ser Arg Ser Ala Ala
1 5 10 15 Val Thr Ser Glu Phe His Leu Val Pro Ser Arg Ser Met Asn
Gly Gln 20 25 30 Pro Leu Thr Cys Val Val Ser His Pro Gly Leu Leu
Gln Asp Gln Arg 35 40 45 Ile Thr His Ile Leu His Val Ser Phe Leu
Ala Glu Ala Ser Val Arg 50 55 60 Gly Leu Glu Asp Gln Asn Leu Trp
His Ile Gly Arg Glu Gly Ala Met 65 70 75 80 Leu Lys Cys Leu Ser Glu
Gly Gln Pro Pro Pro Ser Tyr Asn Trp Thr 85 90 95 Arg Leu Asp Gly
Pro Leu Pro Ser Gly Val Arg Val Asp Gly Asp Thr 100 105 110 Leu Gly
Phe Pro Pro Leu Thr Thr Glu His Ser Gly Ile Tyr Val Cys 115 120 125
His Val Ser Asn Glu Phe Ser Ser Arg Asp Ser Gln Val Thr Val Asp 130
135 140 Val Leu Ala Asp Pro Gln Glu Asp Ser Gly Lys Gln Val Asp Leu
Val 145 150 155 160 Ser Ala Ser Val Val Val Val Gly Val Ile Ala Ala
Leu Leu Phe Cys 165 170 175 Leu Leu Val Val Val Val Val Leu Met Ser
Arg Tyr His Arg Arg Lys 180 185 190 Ala Gln Gln Met Thr Gln Lys Tyr
Glu Glu Glu Leu Thr Leu Thr Arg 195 200 205 Glu Asn Ser Ile Arg Arg
Leu His Ser His His Thr Asp Pro Arg Ser 210 215 220 Gln Ser Glu Glu
Pro Glu Gly Arg Ser Tyr Ser Thr Leu Thr Thr Val 225 230 235 240 Arg
Glu Ile Glu Thr Gln Thr Glu Leu Leu Ser Pro Gly Ser Gly Arg 245 250
255 Ala Glu Glu Glu Glu Asp Gln Asp Glu Gly Ile Lys Gln Ala Met Asn
260 265 270 His Phe Val Gln Glu Asn Gly Thr Leu Arg Ala Lys Pro Thr
Gly Asn 275 280 285 Gly Ile Tyr Ile Asn Gly Arg Gly His Leu Val 290
295 <210> SEQ ID NO 44 <400> SEQUENCE: 44 000 3
<210> SEQ ID NO 45 <400> SEQUENCE: 45 000 3 <210>
SEQ ID NO 46 <400> SEQUENCE: 46 000 3 <210> SEQ ID NO
47 <400> SEQUENCE: 47 000 3 <210> SEQ ID NO 48
<400> SEQUENCE: 48 000 3 <210> SEQ ID NO 49 <400>
SEQUENCE: 49 000 3 <210> SEQ ID NO 50 <400> SEQUENCE:
50 000 3 <210> SEQ ID NO 51 <211> LENGTH: 3114
<212> TYPE: DNA <213> ORGANISM: Homo sapiens
<400> SEQUENCE: 51 cttaatgttg gaagtctctt agtcctatga
gagtgtgtag cagtttgtcc ctgagctcta 60 gcttctttaa atgaagctga
gtctctgggc aacatcttta gggagagagg tacaaaaggt 120 tcctggacct
tctcaacaca gggagcctgc ataatgatgc aagagcagca acctcaaagt 180
acagagaaaa gaggctggtt gtccctgaga ctctggtctg tggctgggat ttccattgca
240 ctcctcagtg cttgcttcat tgtgagctgt gtagtaactt accattttac
atatggtgaa 300 actggcaaaa ggctgtctga actacactca tatcattcaa
gtctcacctg cttcagtgaa 360 gggacaaagg tgccagcctg gggatgttgc
ccagcttctt ggaagtcatt tggttccagt 420 tgctacttca tttccagtga
agagaaggtt tggtctaaga gtgagcagaa ctgtgttgag 480 atgggagcac
atttggttgt gttcaacaca gaagcagagc agaatttcat tgtccagcag 540
ctgaatgagt cattttctta ttttctgggg ctttcagacc cacaaggtaa taataattgg
600 caatggattg ataagacacc ttatgagaaa aatgtcagat tttggcacct
aggtgagccc 660 aatcattctg cagagcaatg tgcttcaata gtcttctgga
aacctacagg atggggctgg 720 aatgatgtta tctgtgaaac tagaaggaat
tcaatatgtg agatgaataa gatttaccta 780 tgagtagaag cttaattgga
aagaagagaa gaattactga cgtaattttt tccctgacgt 840 ctttaaaatt
gaaccctatc atgaaatgat aatttcttcc tgaatttaca cataatcctt 900
atgttataga ggttcacaga aatggaaaga tacctgtttc cctttaatca atcttctcgt
960 ttcctctttt ccattaatga tagaatgcac ccttcctctc tttgttccat
tctttcactt 1020 gttattcatt tttttctttc ttcacacttc attacacaaa
tatttattgt ttcagagact 1080 gtactatttt gtttgttaga agatttataa
ggcagtatct tttgaaaatt atgactttcc 1140 ttcctcaata taccataaag
aaatcttttt ggtcaagatg gtagttggaa ctacaatcat 1200 ctgaaggcct
gacaagagtt gaaagacatg ttttctagat ggctcactca catggctggc 1260
aacttggtgt tggctattaa tgtaacctgg aaataaattt tattctgcag ttagggattt
1320 ggcattttat atatgttgat tcaatcaagt ttggcaagca gggtgttcga
tactgctata 1380 tcctgtattc ttggtttatt tgttttattt ctgagaaata
tgtgttaaga tctctcgctg 1440 attgggaatt tgtctatttc tcatttaaat
tttgtcaaat ctttctttgc ttgcaagcat 1500 ttcttgttac ccaaatctaa
cctattcctg aaaatatgat ggttagcaaa gtttgagata 1560 actagagcct
gtaatccatc attttaaatg gcaatgataa tgacagttta tttttatgtt 1620
atataaaaac ctcaacaaat tttccaaaca attaccaaaa tggtcattaa tctgtatcca
1680 caaaggattt ctgcattaca tactttaaaa caaattacct aattatttag
tgcatattaa 1740 acttattggt gggcatgact atatgcaaca gttgcatgat
atatgataca aattatgtta 1800 ttcttttcca ttgcactgaa aataccataa
tataaagaag aatcccatca tccaaattga 1860 gcctatattg attgatactc
agaagaatct ggcagtagga gcctataaag ggataagcaa 1920 ttgggaaagg
attgggaagt tggtagtact gaacatcttc tcacctggac tcatgagcaa 1980
cttgaatagt tgtaactgtg atgcatatgt agattctaac acatttttcc cccttgaata
2040 gaaatttggc acaacaattt tttaaattaa tttagcaaat atttggatat
taaagcttct 2100 tatagaaaga gatacctgta tatttaagcc atgatgaggt
atatacaatg ttataattat 2160 tacttgtaca tggcaaatta atttttttat
cattgtggag tcactttctt taaatttagt 2220 aatgcctttg gctttaattt
ttctcctgat attaaaatag atacagtaac tttcattatg 2280 ttagtgctgt
aaaatttttt tttccatctt ctatttttga ccatttttat tccacatgtg 2340
ctcttaataa gtagcatata gttaaatttt aaaaaatcca atatggcaat caccttttag
2400 gttaaaaatt taatccattt acatttgtga caattcgaca tatatatggt
tctaaatcta 2460 tcatcttact aggtggtttc catttcctct gctccaaaat
atttttttta cagcttataa 2520 cacaactttt attagaaaag ttatacataa
cacagcatca actattttca agaacccaat 2580 aagcaacaaa aaccagacta
acaaaatgtg taacaagaaa ctaatgacct ttctaaaatc 2640 aaacattcaa
ttatctacaa tgtctattta caaacaggga aaactccatg gtttacaggc 2700
atgtcatatt gaaaataaag ctgcaatagc tttttataca attatcgctc tcaagaaaat
2760 gaatcattaa gacagtaatt aggagttcac aaatttaaaa catttcacgt
aattttaaat 2820 tattgtcttc aataatttta aattattgaa gtctgagttt
caaaagtgat tttttcccac 2880 aaaggtgcca acacttaagc tagagctttc
agtgttaact ttgccctaaa agttaagaca 2940 tattctgaga atcataatag
tcacatgatt tctgatgcta tctgctctgt taataacaaa 3000 gatttcacac
atgaatacct atgtaacaaa tctccatgtt ctacacatat accccagaac 3060
ttaaagtata ataataataa aacatagcaa agcctttaaa aaaaaaaaaa aaaa 3114
<210> SEQ ID NO 52 <211> LENGTH: 627 <212> TYPE:
DNA <213> ORGANISM: Homo sapiens <400> SEQUENCE: 52
atgatgcaag agcagcaacc tcaaagtaca gagaaaagag gctggttgtc cctgagactc
60 tggtctgtgg ctgggatttc cattgcactc ctcagtgctt gcttcattgt
gagctgtgta 120 gtaacttacc attttacata tggtgaaact ggcaaaaggc
tgtctgaact acactcatat 180 cattcaagtc tcacctgctt cagtgaaggg
acaaaggtgc cagcctgggg atgttgccca 240 gcttcttgga agtcatttgg
ttccagttgc tacttcattt ccagtgaaga gaaggtttgg 300 tctaagagtg
agcagaactg tgttgagatg ggagcacatt tggttgtgtt caacacagaa 360
gcagagcaga atttcattgt ccagcagctg aatgagtcat tttcttattt tctggggctt
420 tcagacccac aaggtaataa taattggcaa tggattgata agacacctta
tgagaaaaat 480 gtcagatttt ggcacctagg tgagcccaat cattctgcag
agcaatgtgc ttcaatagtc 540 ttctggaaac ctacaggatg gggctggaat
gatgttatct gtgaaactag aaggaattca 600 atatgtgaga tgaataagat ttaccta
627 <210> SEQ ID NO 53 <211> LENGTH: 209 <212>
TYPE: PRT <213> ORGANISM: Homo sapiens <400> SEQUENCE:
53 Met Met Gln Glu Gln Gln Pro Gln Ser Thr Glu Lys Arg Gly Trp Leu
1 5 10 15 Ser Leu Arg Leu Trp Ser Val Ala Gly Ile Ser Ile Ala Leu
Leu Ser 20 25 30 Ala Cys Phe Ile Val Ser Cys Val Val Thr Tyr His
Phe Thr Tyr Gly 35 40 45 Glu Thr Gly Lys Arg Leu Ser Glu Leu His
Ser Tyr His Ser Ser Leu 50 55 60 Thr Cys Phe Ser Glu Gly Thr Lys
Val Pro Ala Trp Gly Cys Cys Pro 65 70 75 80 Ala Ser Trp Lys Ser Phe
Gly Ser Ser Cys Tyr Phe Ile Ser Ser Glu 85 90 95 Glu Lys Val Trp
Ser Lys Ser Glu Gln Asn Cys Val Glu Met Gly Ala 100 105 110 His Leu
Val Val Phe Asn Thr Glu Ala Glu Gln Asn Phe Ile Val Gln 115 120 125
Gln Leu Asn Glu Ser Phe Ser Tyr Phe Leu Gly Leu Ser Asp Pro Gln 130
135 140 Gly Asn Asn Asn Trp Gln Trp Ile Asp Lys Thr Pro Tyr Glu Lys
Asn 145 150 155 160 Val Arg Phe Trp His Leu Gly Glu Pro Asn His Ser
Ala Glu Gln Cys 165 170 175 Ala Ser Ile Val Phe Trp Lys Pro Thr Gly
Trp Gly Trp Asn Asp Val 180 185 190 Ile Cys Glu Thr Arg Arg Asn Ser
Ile Cys Glu Met Asn Lys Ile Tyr 195 200 205 Leu <210> SEQ ID
NO 54 <211> LENGTH: 48 <212> TYPE: PRT <213>
ORGANISM: Homo sapiens <400> SEQUENCE: 54 Met Met Gln Glu Gln
Gln Pro Gln Ser Thr Glu Lys Arg Gly Trp Leu 1 5 10 15 Ser Leu Arg
Leu Trp Ser Val Ala Gly Ile Ser Ile Ala Leu Leu Ser 20 25 30 Ala
Cys Phe Ile Val Ser Cys Val Val Thr Tyr His Phe Thr Tyr Gly 35 40
45 <210> SEQ ID NO 55 <211> LENGTH: 161 <212>
TYPE: PRT <213> ORGANISM: Homo sapiens <400> SEQUENCE:
55 Glu Thr Gly Lys Arg Leu Ser Glu Leu His Ser Tyr His Ser Ser Leu
1 5 10 15 Thr Cys Phe Ser Glu Gly Thr Lys Val Pro Ala Trp Gly Cys
Cys Pro 20 25 30 Ala Ser Trp Lys Ser Phe Gly Ser Ser Cys Tyr Phe
Ile Ser Ser Glu 35 40 45 Glu Lys Val Trp Ser Lys Ser Glu Gln Asn
Cys Val Glu Met Gly Ala 50 55 60 His Leu Val Val Phe Asn Thr Glu
Ala Glu Gln Asn Phe Ile Val Gln 65 70 75 80 Gln Leu Asn Glu Ser Phe
Ser Tyr Phe Leu Gly Leu Ser Asp Pro Gln 85 90 95 Gly Asn Asn Asn
Trp Gln Trp Ile Asp Lys Thr Pro Tyr Glu Lys Asn 100 105 110 Val Arg
Phe Trp His Leu Gly Glu Pro Asn His Ser Ala Glu Gln Cys 115 120 125
Ala Ser Ile Val Phe Trp Lys Pro Thr Gly Trp Gly Trp Asn Asp Val 130
135 140 Ile Cys Glu Thr Arg Arg Asn Ser Ile Cys Glu Met Asn Lys Ile
Tyr 145 150 155 160 Leu <210> SEQ ID NO 56 <400>
SEQUENCE: 56 000 3 <210> SEQ ID NO 57 <400> SEQUENCE:
57 000 3 <210> SEQ ID NO 58 <400> SEQUENCE: 58 000 3
<210> SEQ ID NO 59 <400> SEQUENCE: 59 000 3 <210>
SEQ ID NO 60 <211> LENGTH: 209 <212> TYPE: PRT
<213> ORGANISM: Mus sp. <400> SEQUENCE: 60 Met Val Gln
Glu Arg Gln Ser Gln Gly Lys Gly Val Cys Trp Thr Leu 1 5 10 15 Arg
Leu Trp Ser Ala Ala Val Ile Ser Met Leu Leu Leu Ser Thr Cys 20 25
30 Phe Ile Ala Ser Cys Val Val Thr Tyr Gln Phe Ile Met Asp Gln Pro
35 40 45 Ser Arg Arg Leu Tyr Glu Leu His Thr Tyr His Ser Ser Leu
Thr Cys 50 55 60 Phe Ser Glu Gly Thr Met Val Ser Glu Lys Met Trp
Gly Cys Cys Pro 65 70 75 80 Asn His Trp Lys Ser Phe Gly Ser Ser Cys
Tyr Leu Ile Ser Thr Lys 85 90 95 Glu Asn Phe Trp Ser Thr Ser Glu
Gln Asn Cys Val Gln Met Gly Ala 100 105 110 His Leu Val Val Ile Asn
Thr Glu Ala Glu Gln Asn Phe Ile Thr Gln 115 120 125 Gln Leu Asn Glu
Ser Leu Ser Tyr Phe Leu Gly Leu Ser Asp Pro Gln 130 135 140 Gly Asn
Gly Lys Trp Gln Trp Ile Asp Asp Thr Pro Phe Ser Gln Asn 145 150 155
160 Val Arg Phe Trp His Pro His Glu Pro Asn Leu Pro Glu Glu Arg Cys
165 170 175 Val Ser Ile Val Tyr Trp Asn Pro Ser Lys Trp Gly Trp Asn
Asp Val 180 185 190 Phe Cys Asp Ser Lys His Asn Ser Ile Cys Glu Met
Lys Lys Ile Tyr 195 200 205 Leu <210> SEQ ID NO 61
<211> LENGTH: 821 <212> TYPE: DNA <213> ORGANISM:
Mus sp. <220> FEATURE: <221> NAME/KEY: misc_feature
<222> LOCATION: (788)..(788) <223> OTHER INFORMATION:
unsure <400> SEQUENCE: 61 gaactccccg gtgtcgaccc cgcgtcccga
ttggcccgct ctgtggcatt taactcaagt 60 gtgtgtggaa gttgattctg
aactctggcc tctttgacag aagccaggtc cctgagtcgt 120 attttggaga
cagatgcaag aaacccctga ccttctgaac atacacctca acaatggtgc 180
aggaaagaca atcccaaggg aagggagtct gctggaccct gagactctgg tcagctgctg
240 tgatttccat gttactcttg agtacctgtt tcattgcgag ctgtgtggtg
acttaccaat 300 ttattatgga ccagcccagt agaagactat atgaacttca
cacataccat tccagtctca 360 cctgcttcag tgaagggact atggtgtcag
aaaaaatgtg gggatgctgc ccaaatcact 420 ggaagtcatt tggctccagc
tgctacctca tttctaccaa ggagaacttc tggagcacca 480 gtgagcagaa
ctgtgttcag atgggggctc atctggtggt gatcaatact gaagcggagc 540
agaatttcat cacccagcag ctgaatgagt cactttctta cttcctgggt ctttcggatc
600 ccaaggtaat ggcaaatggc aatggatcga tgatactcct ttcagtcaaa
atgtcaggtt 660 ctggcacccc catgaaccca atcttccaga agagcggtgt
gtttcaatag tttactggaa 720 tccttcgaaa tggggctggg aatgatgttt
tctgtgatag taaacacaat tcaatatgtg 780 aaatgaanaa gattacctat
gaatgcctgt tattcttaat a 821 <210> SEQ ID NO 62 <211>
LENGTH: 534 <212> TYPE: DNA <213> ORGANISM: Mus sp.
<400> SEQUENCE: 62 atggtgcagg aaagacaatc ccaagggaag
ggagtctgct ggaccctgag actctggtca 60 gctgctgtga tttccatgtt
actcttgagt acctgtttca ttgcgagctg tgtggtgact 120 taccaattta
ttatggacca gcccagtaga agactatatg aacttcacac ataccattcc 180
agtctcacct gcttcagtga agggactatg gtgtcagaaa aaatgtgggg atgctgccca
240 aatcactgga agtcatttgg ctccagctgc tacctcattt ctaccaagga
gaacttctgg 300 agcaccagtg agcagaactg tgttcagatg ggggctcatc
tggtggtgat caatactgaa 360 gcggagcaga atttcatcac ccagcagctg
aatgagtcac tttcttactt cctgggtctt 420 tcggatccca aggtaatggc
aaatggcaat ggatcgatga tactcctttc agtcaaaatg 480 tcaggttctg
gcacccccat gaacccaatc ttccagaaga gcggtgtgtt tcaa 534 <210>
SEQ ID NO 63 <211> LENGTH: 178 <212> TYPE: PRT
<213> ORGANISM: Mus sp. <400> SEQUENCE: 63 Met Val Gln
Glu Arg Gln Ser Gln Gly Lys Gly Val Cys Trp Thr Leu 1 5 10 15 Arg
Leu Trp Ser Ala Ala Val Ile Ser Met Leu Leu Leu Ser Thr Cys 20 25
30 Phe Ile Ala Ser Cys Val Val Thr Tyr Gln Phe Ile Met Asp Gln Pro
35 40 45 Ser Arg Arg Leu Tyr Glu Leu His Thr Tyr His Ser Ser Leu
Thr Cys 50 55 60 Phe Ser Glu Gly Thr Met Val Ser Glu Lys Met Trp
Gly Cys Cys Pro 65 70 75 80 Asn His Trp Lys Ser Phe Gly Ser Ser Cys
Tyr Leu Ile Ser Thr Lys 85 90 95 Glu Asn Phe Trp Ser Thr Ser Glu
Gln Asn Cys Val Gln Met Gly Ala 100 105 110 His Leu Val Val Ile Asn
Thr Glu Ala Glu Gln Asn Phe Ile Thr Gln 115 120 125 Gln Leu Asn Glu
Ser Leu Ser Tyr Phe Leu Gly Leu Ser Asp Pro Lys 130 135 140 Val Met
Ala Asn Gly Asn Gly Ser Met Ile Leu Leu Ser Val Lys Met 145 150 155
160 Ser Gly Ser Gly Thr Pro Met Asn Pro Ile Phe Gln Lys Ser Gly Val
165 170 175 Phe Gln <210> SEQ ID NO 64 <211> LENGTH: 48
<212> TYPE: PRT <213> ORGANISM: Mus sp. <400>
SEQUENCE: 64 Met Val Gln Glu Arg Gln Ser Gln Gly Lys Gly Val Cys
Trp Thr Leu 1 5 10 15 Arg Leu Trp Ser Ala Ala Val Ile Ser Met Leu
Leu Leu Ser Thr Cys 20 25 30 Phe Ile Ala Ser Cys Val Val Thr Tyr
Gln Phe Ile Met Asp Gln Pro 35 40 45 <210> SEQ ID NO 65
<211> LENGTH: 130 <212> TYPE: PRT <213> ORGANISM:
Mus sp. <400> SEQUENCE: 65 Ser Arg Arg Leu Tyr Glu Leu His
Thr Tyr His Ser Ser Leu Thr Cys 1 5 10 15 Phe Ser Glu Gly Thr Met
Val Ser Glu Lys Met Trp Gly Cys Cys Pro 20 25 30 Asn His Trp Lys
Ser Phe Gly Ser Ser Cys Tyr Leu Ile Ser Thr Lys 35 40 45 Glu Asn
Phe Trp Ser Thr Ser Glu Gln Asn Cys Val Gln Met Gly Ala 50 55 60
His Leu Val Val Ile Asn Thr Glu Ala Glu Gln Asn Phe Ile Thr Gln 65
70 75 80 Gln Leu Asn Glu Ser Leu Ser Tyr Phe Leu Gly Leu Ser Asp
Pro Lys 85 90 95 Val Met Ala Asn Gly Asn Gly Ser Met Ile Leu Leu
Ser Val Lys Met 100 105 110 Ser Gly Ser Gly Thr Pro Met Asn Pro Ile
Phe Gln Lys Ser Gly Val 115 120 125 Phe Gln 130 <210> SEQ ID
NO 66 <400> SEQUENCE: 66 000 3 <210> SEQ ID NO 67
<400> SEQUENCE: 67 000 3 <210> SEQ ID NO 68 <400>
SEQUENCE: 68 000 3 <210> SEQ ID NO 69 <400> SEQUENCE:
69 000 3 <210> SEQ ID NO 70 <400> SEQUENCE: 70 000 3
<210> SEQ ID NO 71 <211> LENGTH: 1252 <212> TYPE:
DNA <213> ORGANISM: Mus sp. <400> SEQUENCE: 71
cgaccccgcg tccgctgact tctgggtttg cagcattggc ccgctctgtg gcatttaact
60 caagtgtgtg tggaagttga ttctgaactc tggcctcttt gacagaagcc
aggtccctga 120 gtcgtatttt ggagacagat gcaagaaacc cctgaccttc
tgaacataca cctcaacaat 180 ggtgcaggaa agacaatccc aagggaaggg
agtctgctgg accctgagac tctggtcagc 240 tgctgtgatt tccatgttac
tcttgagtac ctgtttcatt gcgagctgtg tggtgactta 300 ccaatttatt
atggaccagc ccagtagaag actatatgaa cttcacacat accattccag 360
tctcacctgc ttcagtgaag ggactatggt gtcagaaaaa atgtggggat gctgcccaaa
420 tcactggaag tcatttggct ccagctgcta cctcatttct accaaggaga
acttctggag 480 caccagtgag cagaactgtg ttcagatggg ggctcatctg
gtggtgatca atactgaagc 540 ggagcagaat ttcatcaccc agcagctgaa
tgagtcactt tcttacttcc tgggtctttc 600 ggatccacaa ggtaatggca
aatggcaatg gatcgatgat actcctttca gtcaaaatgt 660 caggttctgg
cacccccatg aacccaatct tccagaagag cggtgtgttt caatagttta 720
ctggaatcct tcgaaatggg gctggaatga tgttttctgt gatagtaaac acaattcaat
780 atgtgaaatg aagaagattt acctatgagt gcctgttatt cattaatatc
tttaaagttc 840 agacctacca agaagccata acttcttggc ctgtacatct
gacagaggcc gttcttttcc 900 tagccactat tctttactca aacagaatga
gccctttctc cttctgatgg ttagagtttt 960 gtcaacttga cacaaactag
agtcacctgg ggagtaggat cttcagctaa ggaattgcct 1020 ctgtcagctt
gaccagtcag catgtctggg ggcattttct tgattaatga ttgttgtaag 1080
agggtccagg tggtaagcaa aggtgttaaa cccatgaaga gcaagccagg gagcatcatc
1140 catccatctc tgccctcagg tttctgcccc agggtcttgc cctggtttct
ttctatgaac 1200 tgctgttact tgaaagtata agatgaataa acaatttcat
ccaaaaaaaa aa 1252 <210> SEQ ID NO 72 <211> LENGTH: 627
<212> TYPE: DNA <213> ORGANISM: Mus sp. <400>
SEQUENCE: 72 atggtgcagg aaagacaatc ccaagggaag ggagtctgct ggaccctgag
actctggtca 60 gctgctgtga tttccatgtt actcttgagt acctgtttca
ttgcgagctg tgtggtgact 120 taccaattta ttatggacca gcccagtaga
agactatatg aacttcacac ataccattcc 180 agtctcacct gcttcagtga
agggactatg gtgtcagaaa aaatgtgggg atgctgccca 240 aatcactgga
agtcatttgg ctccagctgc tacctcattt ctaccaagga gaacttctgg 300
agcaccagtg agcagaactg tgttcagatg ggggctcatc tggtggtgat caatactgaa
360 gcggagcaga atttcatcac ccagcagctg aatgagtcac tttcttactt
cctgggtctt 420 tcggatccac aaggtaatgg caaatggcaa tggatcgatg
atactccttt cagtcaaaat 480 gtcaggttct ggcaccccca tgaacccaat
cttccagaag agcggtgtgt ttcaatagtt 540 tactggaatc cttcgaaatg
gggctggaat gatgttttct gtgatagtaa acacaattca 600 atatgtgaaa
tgaagaagat ttaccta 627 <210> SEQ ID NO 73 <211> LENGTH:
586 <212> TYPE: PRT <213> ORGANISM: Mus sp. <400>
SEQUENCE: 73 Met Glu Thr Val Ala Leu Gly Leu Asn Gly Leu Ala Arg
Gly Gly Leu 1 5 10 15 Asn Ser Glu Arg Gly Leu Asn Gly Leu Tyr Leu
Tyr Ser Gly Leu Tyr 20 25 30 Val Ala Leu Cys Tyr Ser Thr Arg Pro
Thr His Arg Leu Glu Ala Arg 35 40 45 Gly Leu Glu Thr Arg Pro Ser
Glu Arg Ala Leu Ala Ala Leu Ala Val 50 55 60 Ala Leu Ile Leu Glu
Ser Glu Arg Met Glu Thr Leu Glu Leu Glu Leu 65 70 75 80 Glu Ser Glu
Arg Thr His Arg Cys Tyr Ser Pro His Glu Ile Leu Glu 85 90 95 Ala
Leu Ala Ser Glu Arg Cys Tyr Ser Val Ala Leu Val Ala Leu Thr 100 105
110 His Arg Thr Tyr Arg Gly Leu Asn Pro His Glu Ile Leu Glu Met Glu
115 120 125 Thr Ala Ser Pro Gly Leu Asn Pro Arg Ser Glu Arg Ala Arg
Gly Ala 130 135 140 Arg Gly Leu Glu Thr Tyr Arg Gly Leu Leu Glu His
Ile Ser Thr His 145 150 155 160 Arg Thr Tyr Arg His Ile Ser Ser Glu
Arg Ser Glu Arg Leu Glu Thr 165 170 175 His Arg Cys Tyr Ser Pro His
Glu Ser Glu Arg Gly Leu Gly Leu Tyr 180 185 190 Thr His Arg Met Glu
Thr Val Ala Leu Ser Glu Arg Gly Leu Leu Tyr 195 200 205 Ser Met Glu
Thr Thr Arg Pro Gly Leu Tyr Cys Tyr Ser Cys Tyr Ser 210 215 220 Pro
Arg Ala Ser Asn His Ile Ser Thr Arg Pro Leu Tyr Ser Ser Glu 225 230
235 240 Arg Pro His Glu Gly Leu Tyr Ser Glu Arg Ser Glu Arg Cys Tyr
Ser 245 250 255 Thr Tyr Arg Leu Glu Ile Leu Glu Ser Glu Arg Thr His
Arg Leu Tyr 260 265 270 Ser Gly Leu Ala Ser Asn Pro His Glu Thr Arg
Pro Ser Glu Arg Thr 275 280 285 His Arg Ser Glu Arg Gly Leu Gly Leu
Asn Ala Ser Asn Cys Tyr Ser 290 295 300 Val Ala Leu Gly Leu Asn Met
Glu Thr Gly Leu Tyr Ala Leu Ala His 305 310 315 320 Ile Ser Leu Glu
Val Ala Leu Val Ala Leu Ile Leu Glu Ala Ser Asn 325 330 335 Thr His
Arg Gly Leu Ala Leu Ala Gly Leu Gly Leu Asn Ala Ser Asn 340 345 350
Pro His Glu Ile Leu Glu Thr His Arg Gly Leu Asn Gly Leu Asn Leu 355
360 365 Glu Ala Ser Asn Gly Leu Ser Glu Arg Leu Glu Ser Glu Arg Thr
Tyr 370 375 380 Arg Pro His Glu Leu Glu Gly Leu Tyr Leu Glu Ser Glu
Arg Ala Ser 385 390 395 400 Pro Pro Arg Gly Leu Asn Gly Leu Tyr Ala
Ser Asn Gly Leu Tyr Leu 405 410 415 Tyr Ser Thr Arg Pro Gly Leu Asn
Thr Arg Pro Ile Leu Glu Ala Ser 420 425 430 Pro Ala Ser Pro Thr His
Arg Pro Arg Pro His Glu Ser Glu Arg Gly 435 440 445 Leu Asn Ala Ser
Asn Val Ala Leu Ala Arg Gly Pro His Glu Thr Arg 450 455 460 Pro His
Ile Ser Pro Arg His Ile Ser Gly Leu Pro Arg Ala Ser Asn 465 470 475
480 Leu Glu Pro Arg Gly Leu Gly Leu Ala Arg Gly Cys Tyr Ser Val Ala
485 490 495 Leu Ser Glu Arg Ile Leu Glu Val Ala Leu Thr Tyr Arg Trp
Ala Ser 500 505 510 Asn Pro Arg Ser Glu Arg Leu Tyr Ser Thr Arg Pro
Gly Leu Tyr Thr 515 520 525 Arg Pro Ala Ser Asn Ala Ser Pro Val Ala
Leu Phe Cys Tyr Ser Ala 530 535 540 Ser Pro Ser Glu Arg Leu Tyr Ser
His Ile Ser Ala Ser Asn Ser Glu 545 550 555 560 Arg Ile Leu Glu Cys
Tyr Ser Gly Leu Met Glu Thr Leu Tyr Ser Leu 565 570 575 Tyr Ser Ile
Leu Glu Thr Tyr Arg Leu Glu 580 585 <210> SEQ ID NO 74
<400> SEQUENCE: 74 000 3 <210> SEQ ID NO 75 <400>
SEQUENCE: 75 000 3 <210> SEQ ID NO 76 <400> SEQUENCE:
76 000 3 <210> SEQ ID NO 77 <400> SEQUENCE: 77 000 3
<210> SEQ ID NO 78 <400> SEQUENCE: 78 000 3 <210>
SEQ ID NO 79 <400> SEQUENCE: 79 000 3 <210> SEQ ID NO
80 <400> SEQUENCE: 80 000 3 <210> SEQ ID NO 81
<211> LENGTH: 1202 <212> TYPE: DNA <213>
ORGANISM: Homo sapiens <400> SEQUENCE: 81 gtcgacccac
gcgtccggaa accattccac aatcaccctc ctgaggaact cttagcactg 60
cataaagtgt tctgagtttg taatcagata ttgtcacact ggttccttca aacagacatg
120 acaaggagct ggctttgggc taggctgctc cttgcctatg attggggaag
gttaaacccc 180 tacagggctt atgtatgtgg aaactgttgg aacactgatt
aaatgggatg gacttcactt 240 aacactcttg gatttccaat attatgtttg
agtaaaagaa ctgctatcca caaacaccat 300 taatccttta gggaggcaga
aaaggccaga atgcaaagcc atcttttcat tacactaggg 360 tctgtctttt
tacttctctg ggcctttatc tggggagggc atgtttcccc cacttggaac 420
agtgagcctg gccaggacag taacctgtgg gcttgtgatg acattatttc taatagggaa
480 tgggaaagga tgttagcttc tcaggtttta aagtgtcctg gaggagaaga
gaaaggacga 540 catgagaagg agacaatgaa gaagatgggt gagggggaga
tagtgtaaga ccctgagaat 600 ggcatagggt aaaactggga cagagatact
gtgggagaac gatagctgca gagggacaga 660 gggaggaagg aaggagaaga
gagggagata aaaacagttt ggagaaactc tcacaataca 720 ttcataagaa
gacaaagaac ccaataaaaa tgggcaacag ataccacaga agatgatata 780
ttgagtggcc aataaataca taaaaatatg ctcaacatct ataattacca gggaaatgca
840 aattaaaagc actgtgagat accactacac actgatgaga atggctaaaa
tcaaaaaaga 900 ccaaccagca ctttgggagg ccgaggtggg cggatcatga
ggtcaggagt ttgagactag 960 cctgaccaac atggtgaaac cctgtctcta
ctaaacatac aaaaattagc tgggggtggt 1020 ggcatgcgcc tgtaattcca
gctactcagg aggctgaggc aggagaatcg cttgaaccca 1080 ggaggcagag
attacagtga gccgagatca tgcccttgca ctctagcctg ggtgacagag 1140
cgagactctg tcttaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aagggcggcc
1200 gc 1202 <210> SEQ ID NO 82 <211> LENGTH: 255
<212> TYPE: DNA <213> ORGANISM: Homo sapiens
<400> SEQUENCE: 82 atgcaaagcc atcttttcat tacactaggg
tctgtctttt tacttctctg ggcctttatc 60 tggggagggc atgtttcccc
cacttggaac agtgagcctg gccaggacag taacctgtgg 120 gcttgtgatg
acattatttc taatagggaa tgggaaagga tgttagcttc tcaggtttta 180
aagtgtcctg gaggagaaga gaaaggacga catgagaagg agacaatgaa gaagatgggt
240 gagggggaga tagtg 255 <210> SEQ ID NO 83 <211>
LENGTH: 85 <212> TYPE: PRT <213> ORGANISM: Homo sapiens
<400> SEQUENCE: 83 Met Gln Ser His Leu Phe Ile Thr Leu Gly
Ser Val Phe Leu Leu Leu 1 5 10 15 Trp Ala Phe Ile Trp Gly Gly His
Val Ser Pro Thr Trp Asn Ser Glu 20 25 30 Pro Gly Gln Asp Ser Asn
Leu Trp Ala Cys Asp Asp Ile Ile Ser Asn 35 40 45 Arg Glu Trp Glu
Arg Met Leu Ala Ser Gln Val Leu Lys Cys Pro Gly 50 55 60 Gly Glu
Glu Lys Gly Arg His Glu Lys Glu Thr Met Lys Lys Met Gly 65 70 75 80
Glu Gly Glu Ile Val 85 <210> SEQ ID NO 84 <211> LENGTH:
23 <212> TYPE: PRT <213> ORGANISM: Homo sapiens
<400> SEQUENCE: 84 Met Gln Ser His Leu Phe Ile Thr Leu Gly
Ser Val Phe Leu Leu Leu 1 5 10 15 Trp Ala Phe Ile Trp Gly Gly 20
<210> SEQ ID NO 85 <211> LENGTH: 62 <212> TYPE:
PRT <213> ORGANISM: Homo sapiens <400> SEQUENCE: 85 His
Val Ser Pro Thr Trp Asn Ser Glu Pro Gly Gln Asp Ser Asn Leu 1 5 10
15 Trp Ala Cys Asp Asp Ile Ile Ser Asn Arg Glu Trp Glu Arg Met Leu
20 25 30 Ala Ser Gln Val Leu Lys Cys Pro Gly Gly Glu Glu Lys Gly
Arg His 35 40 45 Glu Lys Glu Thr Met Lys Lys Met Gly Glu Gly Glu
Ile Val 50 55 60
1 SEQUENCE LISTING <160> NUMBER OF SEQ ID NOS: 85 <210>
SEQ ID NO 1 <211> LENGTH: 2964 <212> TYPE: DNA
<213> ORGANISM: Homo sapiens <400> SEQUENCE: 1
gtcgacccac gcgtccgcgg acgcgtgggg acggctcccg gctgcagtct gcccgcccgc
60 cccgcgcggg ggccgagtcg cgaagcgcgc ctgcgacccg gcgtccgggc
gcgctggaga 120 ggacgcgagg agccatgagg cgccagcctg cgaaggtggc
ggcgctgctg ctcgggctgc 180 tcttggagtg cacagaagcc aaaaagcatt
gctggtattt cgaaggactc tatccaacct 240 attatatatg ccgctcctac
gaggactgct gtggctccag gtgctgtgtg cgggccctct 300 ccatacagag
gctgtggtac ttctggttcc ttctgatgat gggcgtgctt ttctgctgcg 360
gagccggctt cttcatccgg aggcgcatgt accccccgcc gctgatcgag gagccagcct
420 tcaatgtgtc ctacaccagg cagcccccaa atcccggccc aggagcccag
cagccggggc 480 cgccctatta cactgaccca ggaggaccgg ggatgaaccc
tgtcgggaat tccatggcaa 540 tggctttcca ggtcccaccc aactcacccc
aggggagtgt ggcctgcccg ccccctccag 600 cctactgcaa cacgcctccg
cccccgtacg aacaggtagt gaaggccaag tagtggggtg 660 cccacgtgca
agaggagaga caggagaggg cctttccctg gcctttctgt cttcgttgat 720
gttcacttcc aggaacggtc tcgtgggctg ctaagggcag ttcctctgat atcctcacag
780 caagcacagc tctctttcag gctttccatg gagtacaata tatgaactca
cactttgtct 840 cctctgttgc ttctgtttct gacgcagtct gtgctctcac
atggtagtgt ggtgacagtc 900 cccgagggct gacgtcctta cggtggcgtg
accagatcta caggagagag actgagagga 960 agaaggcagt gctggaggtg
caggtggcat gtagaggggc caggccgagc atcccaggca 1020 agcatccttc
tgcccgggta ttaataggaa gccccatgcc gggcggctca gccgatgaag 1080
cagcagccga ctgagctgag cccagcaggt catctgctcc agcctgtcct ctcgtcagcc
1140 ttcctcttcc agaagctgtt ggagagacat tcaggagaga gcaagcccct
tgtcatgttt 1200 ctgtctctgt tcatatccta aagatagact tctcctgcac
cgccagggaa gggtagcacg 1260 tgcagctctc accgcaggat ggggcctaga
atcaggcttg ccttggaggc ctgacagtga 1320 tctgacatcc actaagcaaa
tttatttaaa ttcatgggaa atcacttcct gccccaaact 1380 gagacattgc
attttgtgag ctcttggtct gatttggaga aaggactgtt acccattttt 1440
ttggtgtgtt tatggaagtg catgtagagc gtcctgccct ttgaaatcag actgggtgtg
1500 tgtcttccct ggacatcact gcctctccag ggcattctca ggcccggggg
tctccttccc 1560 tcaggcagct ccagtggtgg gttctgaagg gtgctttcaa
aacggggcac atctggctgg 1620 gaagtcacat ggactcttcc agggagagag
accagctgag gcgtctctct ctgaggttgt 1680 gttgggtcta agcgggtgtg
tgctgggctc caaggaggag gagcttgctg ggaaaagaca 1740 ggagaagtac
tgactcaact gcactgacca tgttgtcata attagaataa agaagaagtg 1800
gtcggaaatg cacattcctg gataggaatc acagctcacc ccaggatctc acaggtagtc
1860 tcctgagtag ttgacggcta gcggggagct agttccgccg catagttata
gtgttgatgt 1920 gtgaacgctg acctgtcctg tgtgctaaga gctatgcagc
ttagctgagg cgcctagatt 1980 actagatgtg ctgtatcacg gggaatgagg
tgggggtgct tattttttaa tgaactaatc 2040 agagcctctt gagaaattgt
tactcattga actggagcat caagacatct catggaagtg 2100 gatacggagt
gatttggtgt ccatgctttt cactctgagg acatttaatc ggagaacctc 2160
ctggggaatt ttgtgggaga cacttgggaa caaaacagac accctgggaa tgcagttgca
2220 agcacagatg ctgccaccag tgtctctgac caccctggtg tgactgctga
ctgccagcgt 2280 ggtacctccc atgctgcagg cctccatcta aatgagacaa
caaagcacaa tgttcactgt 2340 ttacaaccaa gacaactgcg tgggtccaaa
cactcctctt cctccaggtc atttgttttg 2400 catttttaat gtctttattt
tttgtaatga aaaagcacac taagctgccc ctggaatcgg 2460 gtgcagctga
ataggcaccc aaaagtccgt gactaaattt cgtttgtctt tttgatagca 2520
aattatgtta agagacagtg atggctaggg ctcaacaatt ttgtattccc atgtttgtgt
2580 gagacagagt ttgttttccc ttgaacttgg ttagaattgt gctactgtga
acgctgatcc 2640 tgcatatgga agtcccactt tggtgacatt tcctggccat
tcttgtttcc attgtgtgga 2700 tggtgggttg tgcccacttc ctggagtgag
acagctcctg gtgtgtagaa ttcccggagc 2760 gtccgtggtt cagagtaaac
ttgaagcaga tctgtgcatg cttttcctct gcaacaattg 2820 gctcgtttct
cttttttgtt ctcttttgat aggatcctgt ttcctatgtg tgcaaaataa 2880
aaataaattt gggcaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa
2940 aaaaaaaaaa aaaagggcgg ccgc 2964 <210> SEQ ID NO 2
<211> LENGTH: 516 <212> TYPE: DNA <213> ORGANISM:
Homo sapiens <400> SEQUENCE: 2 atgaggcgcc agcctgcgaa
ggtggcggcg ctgctgctcg ggctgctctt ggagtgcaca 60 gaagccaaaa
agcattgctg gtatttcgaa ggactctatc caacctatta tatatgccgc 120
tcctacgagg actgctgtgg ctccaggtgc tgtgtgcggg ccctctccat acagaggctg
180 tggtacttct ggttccttct gatgatgggc gtgcttttct gctgcggagc
cggcttcttc 240 atccggaggc gcatgtaccc cccgccgctg atcgaggagc
cagccttcaa tgtgtcctac 300 accaggcagc ccccaaatcc cggcccagga
gcccagcagc cggggccgcc ctattacact 360 gacccaggag gaccggggat
gaaccctgtc gggaattcca tggcaatggc tttccaggtc 420 ccacccaact
caccccaggg gagtgtggcc tgcccgcccc ctccagccta ctgcaacacg 480
cctccgcccc cgtacgaaca ggtagtgaag gccaag 516 <210> SEQ ID NO 3
<211> LENGTH: 172 <212> TYPE: PRT <213> ORGANISM:
Homo sapiens <400> SEQUENCE: 3 Met Arg Arg Gln Pro Ala Lys
Val Ala Ala Leu Leu Leu Gly Leu Leu 1 5 10 15 Leu Glu Cys Thr Glu
Ala Lys Lys His Cys Trp Tyr Phe Glu Gly Leu 20 25 30 Tyr Pro Thr
Tyr Tyr Ile Cys Arg Ser Tyr Glu Asp Cys Cys Gly Ser 35 40 45 Arg
Cys Cys Val Arg Ala Leu Ser Ile Gln Arg Leu Trp Tyr Phe Trp 50 55
60 Phe Leu Leu Met Met Gly Val Leu Phe Cys Cys Gly Ala Gly Phe Phe
65 70 75 80 Ile Arg Arg Arg Met Tyr Pro Pro Pro Leu Ile Glu Glu Pro
Ala Phe 85 90 95 Asn Val Ser Tyr Thr Arg Gln Pro Pro Asn Pro Gly
Pro Gly Ala Gln 100 105 110 Gln Pro Gly Pro Pro Tyr Tyr Thr Asp Pro
Gly Gly Pro Gly Met Asn 115 120 125 Pro Val Gly Asn Ser Met Ala Met
Ala Phe Gln Val Pro Pro Asn Ser 130 135 140 Pro Gln Gly Ser Val Ala
Cys Pro Pro Pro Pro Ala Tyr Cys Asn Thr 145 150 155 160 Pro Pro Pro
Pro Tyr Glu Gln Val Val Lys Ala Lys 165 170 <210> SEQ ID NO 4
<211> LENGTH: 22 <212> TYPE: PRT <213> ORGANISM:
Homo sapiens <400> SEQUENCE: 4 Met Arg Arg Gln Pro Ala Lys
Val Ala Ala Leu Leu Leu Gly Leu Leu 1 5 10 15 Leu Glu Cys Thr Glu
Ala 20 <210> SEQ ID NO 5 <211> LENGTH: 150 <212>
TYPE: PRT <213> ORGANISM: Homo sapiens <400> SEQUENCE:
5 Lys Lys His Cys Trp Tyr Phe Glu Gly Leu Tyr Pro Thr Tyr Tyr Ile 1
5 10 15 Cys Arg Ser Tyr Glu Asp Cys Cys Gly Ser Arg Cys Cys Val Arg
Ala 20 25 30 Leu Ser Ile Gln Arg Leu Trp Tyr Phe Trp Phe Leu Leu
Met Met Gly 35 40 45 Val Leu Phe Cys Cys Gly Ala Gly Phe Phe Ile
Arg Arg Arg Met Tyr 50 55 60 Pro Pro Pro Leu Ile Glu Glu Pro Ala
Phe Asn Val Ser Tyr Thr Arg 65 70 75 80 Gln Pro Pro Asn Pro Gly Pro
Gly Ala Gln Gln Pro Gly Pro Pro Tyr 85 90 95 Tyr Thr Asp Pro Gly
Gly Pro Gly Met Asn Pro Val Gly Asn Ser Met 100 105 110 Ala Met Ala
Phe Gln Val Pro Pro Asn Ser Pro Gln Gly Ser Val Ala 115 120 125 Cys
Pro Pro Pro Pro Ala Tyr Cys Asn Thr Pro Pro Pro Pro Tyr Glu 130 135
140 Gln Val Val Lys Ala Lys 145 150 <210> SEQ ID NO 6
<211> LENGTH: 38 <212> TYPE: PRT <213> ORGANISM:
Homo sapiens <400> SEQUENCE: 6 Lys Lys His Cys Trp Tyr Phe
Glu Gly Leu Tyr Pro Thr Tyr Tyr Ile 1 5 10 15 Cys Arg Ser Tyr Glu
Asp Cys Cys Gly Ser Arg Cys Cys Val Arg Ala 20 25 30 Leu Ser Ile
Gln Arg Leu 35
<210> SEQ ID NO 7 <211> LENGTH: 21 <212> TYPE:
PRT <213> ORGANISM: Homo sapiens <400> SEQUENCE: 7 Trp
Tyr Phe Trp Phe Leu Leu Met Met Gly Val Leu Phe Cys Cys Gly 1 5 10
15 Ala Gly Phe Phe Ile 20 <210> SEQ ID NO 8 <211>
LENGTH: 91 <212> TYPE: PRT <213> ORGANISM: Homo sapiens
<400> SEQUENCE: 8 Arg Arg Arg Met Tyr Pro Pro Pro Leu Ile Glu
Glu Pro Ala Phe Asn 1 5 10 15 Val Ser Tyr Thr Arg Gln Pro Pro Asn
Pro Gly Pro Gly Ala Gln Gln 20 25 30 Pro Gly Pro Pro Tyr Tyr Thr
Asp Pro Gly Gly Pro Gly Met Asn Pro 35 40 45 Val Gly Asn Ser Met
Ala Met Ala Phe Gln Val Pro Pro Asn Ser Pro 50 55 60 Gln Gly Ser
Val Ala Cys Pro Pro Pro Pro Ala Tyr Cys Asn Thr Pro 65 70 75 80 Pro
Pro Pro Tyr Glu Gln Val Val Lys Ala Lys 85 90 <210> SEQ ID NO
9 <211> LENGTH: <212> TYPE: <213> ORGANISM:
<400> SEQUENCE: 9 000 <210> SEQ ID NO 10 <211>
LENGTH: <212> TYPE: <213> ORGANISM: <400>
SEQUENCE: 10 000 <210> SEQ ID NO 11 <211> LENGTH: 2915
<212> TYPE: DNA <213> ORGANISM: Mus sp. <400>
SEQUENCE: 11 gtcgacccac gcgtccggcc gcgcgtcctt ctgccggctt cagctcgtat
ccccggagtc 60 cacccgcccg tcccggggtg cggactggcc ctgagctggc
cgtacagccc ggcttcggac 120 ggtcctcgct ggagccatgg gccgccggct
cggcagggtg gcggcgctgc tgctcgggct 180 gctagtggag tgcactgagg
ccaaaaaaca ttgctggtat tttgaaggac tctatcccac 240 atactatata
tgccgttcct atgaagactg ctgtggctcc aggtgctgtg tgagggccct 300
ttccatacag aggctgtggt atttttggtt cctgctgatg atgggtgtgc tgttctgctg
360 tggtgccggt ttcttcattc gccggcgcat gtatccgcca ccactcattg
aggagcccac 420 attcaatgtg tcctatacca ggcagccacc aaatcctgct
ccaggagcac agcaaatggg 480 accgccatat tacaccgacc ctggaggacc
cgggatgaat cctgttggca ataccatggc 540 tatggctttc caggtccagc
ccaattcacc tcacggaggc acaacttacc caccccctcc 600 ttcctactgc
aacacgcctc caccccccta tgaacaggtg gtgaaggaca agtagcaaga 660
tgctacatca aaggcaaaga ggatggacag gcccttttgt ttaccttccc atcctcaccg
720 atacttgctg atagggtggt ccaagggaaa acttggatat tctcaaagca
agcccagctc 780 tctttcaagt cttttgtgga ggacatttga atccacactg
tctcctctgt tgcttctgtt 840 tctgatgtag tctgtgctct ctgagagagt
gtggcaacag tccctgaggg ttgatattcc 900 tagggtgtcc agggtagatc
ctcgggagag aggctaaggg gaaaggaagg catagcctgt 960 gtgttagggg
gcagataaag tggtcaggct gagataagac tcacatgatg cagtagttgg 1020
cagtgaactt cgaagagaca ctatccacca tcccagccca ttctcctaat agaagctgtg
1080 gggctgtgtt gttgatgctc tttggtctcc actcacattt tgaaaatagg
ctttcctctg 1140 caggaatagg aaagacccaa gtacatattt gcttccactt
aaaaatgagg gtcagaacca 1200 ggcctcagtt ggacatctat agttaaataa
aggccattag agaggggaaa tctttaagtt 1260 aggggaaatt ctctaaatgg
agacattgcg ttttatgaat catcgtctgg cttttctttt 1320 agtgcatgta
ttgaagtgag ggtgtccttt gagatcagat ggggagagtg aactctgcgg 1380
ggggtggggt gtctctactc agagggctcc aacacccttt tcttaggtag ttctggtgat
1440 gggttttatg ggcactatag agctgagggg cacattaggc cgggtagtta
cattgaccct 1500 tggagaggaa gaggacagcc aaagaaactc agcaaagcaa
gaccagcatt gctgagttag 1560 agctagggtt gtatgtgatc ccaacagaga
tgtgctggcc tcagaagagg ggacgtttgt 1620 ggatagagcc gtgaaaacct
acttagttgc acagatgaca taatcaaaag tagagaaaga 1680 agtgtagtta
gagatgccat ttcccaggtg agaatcagag ctcatccata gatttacaag 1740
tagtggctgg agttaacagt atggagttct tttcccttgc gtagttagtc acgttgatgt
1800 gtatttaaac ccaggttgag accttgtgta ctaagagcaa ggaagtatag
ctaagatgtc 1860 tagattattt atatgtagta tggtggggag tggggctgca
aggaaggggg ctgacattgt 1920 aaatgagaaa atcagagcca tttgataaac
tgttacttgt tggatcaggc atccaaaagt 1980 gtctcttgag tggacattga
gtattcttta ccacctacaa gaccaggagg catggtgtca 2040 ttctccattg
gggtatttat atgaggtaga ggttcaggaa tcgacagtag ctgtgtgggc 2100
ttagtttaag gactgaaagc atagggactg gtagacagtt tcataggaaa ctgcggggaa
2160 ggaatggata cctttaaaga cagtttgtgg atgcagatgc tgccacccat
cattgagcac 2220 ccttgtgtct ctggcttcct gtcactggat ccagtacccc
tccatgcttg ggtccttgtt 2280 ttacataaga caacaaagca caatgtctgc
tgtttacaat caagacgact acatggtcca 2340 aacatttctt ctctcttcta
tcacttgtgg ctttaacttc catttcctcc gttccttttt 2400 aaaatcaaga
agcacagtca gagctgcccc tgggattgca tcagggaacg gctgatcaag 2460
gcattcagtg tccatgacta aatcttatct ttttgatagc aaatcctttt aagaaactga
2520 acaattgcta aggctcagca attttatact ccaatgtctg tgtaaggtaa
attttgtttg 2580 ccattgagcc cacattggaa ttccttctga cgtcaacact
gacaatgcct atggaaattg 2640 cacttctggg tatatgtccc agcatccttg
ttttcttatg tttggtgagt aaggctcacc 2700 ccttccagca gctctacttc
tgtgtgctga ggtcctgtag agccggggct tgggcacaga 2760 catgaggcag
acttgtgcat gctctttctt ggcaacactt ggctcatatt tcttgttctc 2820
ttttgataga gtcctgtttc ctatgtattt aaaaaataat aaaagtgaat ttagtcaaaa
2880 aaaaaaaaaa aaaaaaaaaa aaaaagggcg gccgc 2915 <210> SEQ ID
NO 12 <211> LENGTH: 516 <212> TYPE: DNA <213>
ORGANISM: Mus sp. <400> SEQUENCE: 12 atgggccgcc ggctcggcag
ggtggcggcg ctgctgctcg ggctgctagt ggagtgcact 60 gaggccaaaa
aacattgctg gtattttgaa ggactctatc ccacatacta tatatgccgt 120
tcctatgaag actgctgtgg ctccaggtgc tgtgtgaggg ccctttccat acagaggctg
180 tggtattttt ggttcctgct gatgatgggt gtgctgttct gctgtggtgc
cggtttcttc 240 attcgccggc gcatgtatcc gccaccactc attgaggagc
ccacattcaa tgtgtcctat 300 accaggcagc caccaaatcc tgctccagga
gcacagcaaa tgggaccgcc atattacacc 360 gaccctggag gacccgggat
gaatcctgtt ggcaatacca tggctatggc tttccaggtc 420 cagcccaatt
cacctcacgg aggcacaact tacccacccc ctccttccta ctgcaacacg 480
cctccacccc cctatgaaca ggtggtgaag gacaag 516 <210> SEQ ID NO
13 <211> LENGTH: 172 <212> TYPE: PRT <213>
ORGANISM: Mus sp. <400> SEQUENCE: 13 Met Gly Arg Arg Leu Gly
Arg Val Ala Ala Leu Leu Leu Gly Leu Leu 1 5 10 15 Val Glu Cys Thr
Glu Ala Lys Lys His Cys Trp Tyr Phe Glu Gly Leu 20 25 30 Tyr Pro
Thr Tyr Tyr Ile Cys Arg Ser Tyr Glu Asp Cys Cys Gly Ser 35 40 45
Arg Cys Cys Val Arg Ala Leu Ser Ile Gln Arg Leu Trp Tyr Phe Trp 50
55 60 Phe Leu Leu Met Met Gly Val Leu Phe Cys Cys Gly Ala Gly Phe
Phe 65 70 75 80 Ile Arg Arg Arg Met Tyr Pro Pro Pro Leu Ile Glu Glu
Pro Thr Phe 85 90 95 Asn Val Ser Tyr Thr Arg Gln Pro Pro Asn Pro
Ala Pro Gly Ala Gln 100 105 110 Gln Met Gly Pro Pro Tyr Tyr Thr Asp
Pro Gly Gly Pro Gly Met Asn 115 120 125 Pro Val Gly Asn Thr Met Ala
Met Ala Phe Gln Val Gln Pro Asn Ser 130 135 140 Pro His Gly Gly Thr
Thr Tyr Pro Pro Pro Pro Ser Tyr Cys Asn Thr 145 150 155 160 Pro Pro
Pro Pro Tyr Glu Gln Val Val Lys Asp Lys 165 170 <210> SEQ ID
NO 14 <211> LENGTH: <212> TYPE: <213> ORGANISM:
<400> SEQUENCE: 14 000 <210> SEQ ID NO 15 <211>
LENGTH: 150 <212> TYPE: PRT <213> ORGANISM: Mus sp.
<400> SEQUENCE: 15 Lys Lys His Cys Trp Tyr Phe Glu Gly Leu
Tyr Pro Thr Tyr Tyr Ile
1 5 10 15 Cys Arg Ser Tyr Glu Asp Cys Cys Gly Ser Arg Cys Cys Val
Arg Ala 20 25 30 Leu Ser Ile Gln Arg Leu Trp Tyr Phe Trp Phe Leu
Leu Met Met Gly 35 40 45 Val Leu Phe Cys Cys Gly Ala Gly Phe Phe
Ile Arg Arg Arg Met Tyr 50 55 60 Pro Pro Pro Leu Ile Glu Glu Pro
Thr Phe Asn Val Ser Tyr Thr Arg 65 70 75 80 Gln Pro Pro Asn Pro Ala
Pro Gly Ala Gln Gln Met Gly Pro Pro Tyr 85 90 95 Tyr Thr Asp Pro
Gly Gly Pro Gly Met Asn Pro Val Gly Asn Thr Met 100 105 110 Ala Met
Ala Phe Gln Val Gln Pro Asn Ser Pro His Gly Gly Thr Thr 115 120 125
Tyr Pro Pro Pro Pro Ser Tyr Cys Asn Thr Pro Pro Pro Pro Tyr Glu 130
135 140 Gln Val Val Lys Asp Lys 145 150 <210> SEQ ID NO 16
<400> SEQUENCE: 16 000 3 <210> SEQ ID NO 17 <400>
SEQUENCE: 17 000 3 <210> SEQ ID NO 18 <400> SEQUENCE:
18 000 3 <210> SEQ ID NO 19 <211> LENGTH: <212>
TYPE: <213> ORGANISM: <400> SEQUENCE: 19 000
<210> SEQ ID NO 20 <400> SEQUENCE: 20 000 3 <210>
SEQ ID NO 21 <211> LENGTH: 2169 <212> TYPE: DNA
<213> ORGANISM: Homo sapiens <400> SEQUENCE: 21
gtcgacccac gcgtccggaa atgtcgttct tcagatttaa aaagaaaacc tttactgaat
60 cagctgagtg ttaataatac gaatttcctt ttcttgccaa ttctgatctg
aacagaaaat 120 ccaagaacag ggatatgtgt ggattacagt tttctctgcc
ttgcctacga ctgtttctgg 180 ttgttacctg ttatctttta ttattactcc
acaaagaaat acttggatgt tcgtctgttt 240 gtcagctctg cactgggaga
caaattaact gccgtaactt aggcctttcg agtattccta 300 agaattttcc
tgaaagtaca gtttttctgt atctgactgg gaataatata tcttatataa 360
atgaaagtga attaacagga cttcattctc ttgtagcatt gtatttggat aattctaaca
420 ttctgtatgt atatccaaaa gcctttgttc aattgaggca tctatatttt
ctatttctaa 480 ataataattt catcaaacgc ttagatcctg gaatatttaa
gggactttta aatcttcgta 540 atttatattt acagtataat caggtatctt
ttgttccgag aggagtattt aatgatctag 600 tttcagttca gtacttaaat
ctacaaagga atcgcctcac tgtccttggg agtggtacct 660 ttgttggtat
ggttgctctt cggatacttg atttatcaaa caataacatt ttgaggatat 720
cagaatcagg ctttcaacat cttgaaaacc ttgcttgttt gtatttagga agtaataatt
780 taacaaaagt accatcaaat gcctttgaag tacttaaaag tcttagaaga
ctttctttgt 840 ctcataatcc tattgaagca atacagccct ttgcatttaa
aggacttgcc aatctggaat 900 acctcctcct gaaaaattca agaattagga
atgttactag ggatgggttt agtggaatta 960 ataatcttaa acatttgatc
ttaagtcata atgatttaga gaatttaaat tctgacacat 1020 tcagtttgtt
aaagaattta atttacctta agttagatag aaacagaata attagcattg 1080
ataatgatac atttgaaaat atgggagcat ctttgaagat ccttaatctg tcatttaata
1140 atcttacagc cttgcatcca agggtcctta agccgttgtc ttcattgatt
catcttcagg 1200 caaattctaa tccttgggaa tgtaactgca aacttttggg
ccttcgagac tggctagcat 1260 cttcagccat tactctaaac atctattgtc
agaatccccc atccatgcgt ggcagagcat 1320 tacgttatat taacattaca
aattgtgtta catcttcaat aaatgtatcc agagcttggg 1380 ctgttgtaaa
atctcctcat attcatcaca agactactgc gctaatgatg gcctggcata 1440
aagtaaccac aaatggcagt cctctggaaa atactgagac tgagaacatt actttctggg
1500 aacgaattcc tacttcacct gctggtagat tttttcaaga gaatgccttt
ggtaatccat 1560 tagagactac agcagtgtta cctgtgcaaa tacaacttac
tacttctgtt accttgaact 1620 tggaaaaaaa cagtgctcta ccgaatgatg
ctgcttcaat gtcagggaaa acatctctaa 1680 tttgtacaca agaagttgag
aagttgaatg aggcttttga cattttgcta gcttttttca 1740 tcttagcttg
tgttttaatc atttttttga tctacaaagt tgttcagttt aaacaaaaac 1800
taaaggcatc agaaaactca agggaaaata gacttgaata ctacagcttt tatcagtcag
1860 caaggtataa tgtaactgcc tcaatttgta acacttcccc aaattctcta
gaaagtcctg 1920 gcttggagca gattcgactt cataaacaaa ttgttcctga
aaatgaggca caggtcattc 1980 tttttgaaca ttctgcttta taactcaact
aaatattgtc tataagaaac ttcagtgcca 2040 tggacatgat ttaaactgaa
acctccttat ataattatat actttagttg gaaatataat 2100 gaattatatg
aggttagcat tattaaaata tgtttttaat aaaaaaaaaa aaaaaaaaag 2160
ggcggccgc 2169 <210> SEQ ID NO 22 <211> LENGTH: 1866
<212> TYPE: DNA <213> ORGANISM: Homo sapiens
<400> SEQUENCE: 22 atgtgtggat tacagttttc tctgccttgc
ctacgactgt ttctggttgt tacctgttat 60 cttttattat tactccacaa
agaaatactt ggatgttcgt ctgtttgtca gctctgcact 120 gggagacaaa
ttaactgccg taacttaggc ctttcgagta ttcctaagaa ttttcctgaa 180
agtacagttt ttctgtatct gactgggaat aatatatctt atataaatga aagtgaatta
240 acaggacttc attctcttgt agcattgtat ttggataatt ctaacattct
gtatgtatat 300 ccaaaagcct ttgttcaatt gaggcatcta tattttctat
ttctaaataa taatttcatc 360 aaacgcttag atcctggaat atttaaggga
cttttaaatc ttcgtaattt atatttacag 420 tataatcagg tatcttttgt
tccgagagga gtatttaatg atctagtttc agttcagtac 480 ttaaatctac
aaaggaatcg cctcactgtc cttgggagtg gtacctttgt tggtatggtt 540
gctcttcgga tacttgattt atcaaacaat aacattttga ggatatcaga atcaggcttt
600 caacatcttg aaaaccttgc ttgtttgtat ttaggaagta ataatttaac
aaaagtacca 660 tcaaatgcct ttgaagtact taaaagtctt agaagacttt
ctttgtctca taatcctatt 720 gaagcaatac agccctttgc atttaaagga
cttgccaatc tggaatacct cctcctgaaa 780 aattcaagaa ttaggaatgt
tactagggat gggtttagtg gaattaataa tcttaaacat 840 ttgatcttaa
gtcataatga tttagagaat ttaaattctg acacattcag tttgttaaag 900
aatttaattt accttaagtt agatagaaac agaataatta gcattgataa tgatacattt
960 gaaaatatgg gagcatcttt gaagatcctt aatctgtcat ttaataatct
tacagccttg 1020 catccaaggg tccttaagcc gttgtcttca ttgattcatc
ttcaggcaaa ttctaatcct 1080 tgggaatgta actgcaaact tttgggcctt
cgagactggc tagcatcttc agccattact 1140 ctaaacatct attgtcagaa
tcccccatcc atgcgtggca gagcattacg ttatattaac 1200 attacaaatt
gtgttacatc ttcaataaat gtatccagag cttgggctgt tgtaaaatct 1260
cctcatattc atcacaagac tactgcgcta atgatggcct ggcataaagt aaccacaaat
1320 ggcagtcctc tggaaaatac tgagactgag aacattactt tctgggaacg
aattcctact 1380 tcacctgctg gtagattttt tcaagagaat gcctttggta
atccattaga gactacagca 1440 gtgttacctg tgcaaataca acttactact
tctgttacct tgaacttgga aaaaaacagt 1500 gctctaccga atgatgctgc
ttcaatgtca gggaaaacat ctctaatttg tacacaagaa 1560 gttgagaagt
tgaatgaggc ttttgacatt ttgctagctt ttttcatctt agcttgtgtt 1620
ttaatcattt ttttgatcta caaagttgtt cagtttaaac aaaaactaaa ggcatcagaa
1680 aactcaaggg aaaatagact tgaatactac agcttttatc agtcagcaag
gtataatgta 1740 actgcctcaa tttgtaacac ttccccaaat tctctagaaa
gtcctggctt ggagcagatt 1800 cgacttcata aacaaattgt tcctgaaaat
gaggcacagg tcattctttt tgaacattct 1860 gcttta 1866 <210> SEQ
ID NO 23 <211> LENGTH: 622 <212> TYPE: PRT <213>
ORGANISM: Homo sapiens <400> SEQUENCE: 23 Met Cys Gly Leu Gln
Phe Ser Leu Pro Cys Leu Arg Leu Phe Leu Val 1 5 10 15 Val Thr Cys
Tyr Leu Leu Leu Leu Leu His Lys Glu Ile Leu Gly Cys 20 25 30 Ser
Ser Val Cys Gln Leu Cys Thr Gly Arg Gln Ile Asn Cys Arg Asn 35 40
45 Leu Gly Leu Ser Ser Ile Pro Lys Asn Phe Pro Glu Ser Thr Val Phe
50 55 60 Leu Tyr Leu Thr Gly Asn Asn Ile Ser Tyr Ile Asn Glu Ser
Glu Leu 65 70 75 80 Thr Gly Leu His Ser Leu Val Ala Leu Tyr Leu Asp
Asn Ser Asn Ile 85 90 95 Leu Tyr Val Tyr Pro Lys Ala Phe Val Gln
Leu Arg His Leu Tyr Phe 100 105 110 Leu Phe Leu Asn Asn Asn Phe Ile
Lys Arg Leu Asp Pro Gly Ile Phe
115 120 125 Lys Gly Leu Leu Asn Leu Arg Asn Leu Tyr Leu Gln Tyr Asn
Gln Val 130 135 140 Ser Phe Val Pro Arg Gly Val Phe Asn Asp Leu Val
Ser Val Gln Tyr 145 150 155 160 Leu Asn Leu Gln Arg Asn Arg Leu Thr
Val Leu Gly Ser Gly Thr Phe 165 170 175 Val Gly Met Val Ala Leu Arg
Ile Leu Asp Leu Ser Asn Asn Asn Ile 180 185 190 Leu Arg Ile Ser Glu
Ser Gly Phe Gln His Leu Glu Asn Leu Ala Cys 195 200 205 Leu Tyr Leu
Gly Ser Asn Asn Leu Thr Lys Val Pro Ser Asn Ala Phe 210 215 220 Glu
Val Leu Lys Ser Leu Arg Arg Leu Ser Leu Ser His Asn Pro Ile 225 230
235 240 Glu Ala Ile Gln Pro Phe Ala Phe Lys Gly Leu Ala Asn Leu Glu
Tyr 245 250 255 Leu Leu Leu Lys Asn Ser Arg Ile Arg Asn Val Thr Arg
Asp Gly Phe 260 265 270 Ser Gly Ile Asn Asn Leu Lys His Leu Ile Leu
Ser His Asn Asp Leu 275 280 285 Glu Asn Leu Asn Ser Asp Thr Phe Ser
Leu Leu Lys Asn Leu Ile Tyr 290 295 300 Leu Lys Leu Asp Arg Asn Arg
Ile Ile Ser Ile Asp Asn Asp Thr Phe 305 310 315 320 Glu Asn Met Gly
Ala Ser Leu Lys Ile Leu Asn Leu Ser Phe Asn Asn 325 330 335 Leu Thr
Ala Leu His Pro Arg Val Leu Lys Pro Leu Ser Ser Leu Ile 340 345 350
His Leu Gln Ala Asn Ser Asn Pro Trp Glu Cys Asn Cys Lys Leu Leu 355
360 365 Gly Leu Arg Asp Trp Leu Ala Ser Ser Ala Ile Thr Leu Asn Ile
Tyr 370 375 380 Cys Gln Asn Pro Pro Ser Met Arg Gly Arg Ala Leu Arg
Tyr Ile Asn 385 390 395 400 Ile Thr Asn Cys Val Thr Ser Ser Ile Asn
Val Ser Arg Ala Trp Ala 405 410 415 Val Val Lys Ser Pro His Ile His
His Lys Thr Thr Ala Leu Met Met 420 425 430 Ala Trp His Lys Val Thr
Thr Asn Gly Ser Pro Leu Glu Asn Thr Glu 435 440 445 Thr Glu Asn Ile
Thr Phe Trp Glu Arg Ile Pro Thr Ser Pro Ala Gly 450 455 460 Arg Phe
Phe Gln Glu Asn Ala Phe Gly Asn Pro Leu Glu Thr Thr Ala 465 470 475
480 Val Leu Pro Val Gln Ile Gln Leu Thr Thr Ser Val Thr Leu Asn Leu
485 490 495 Glu Lys Asn Ser Ala Leu Pro Asn Asp Ala Ala Ser Met Ser
Gly Lys 500 505 510 Thr Ser Leu Ile Cys Thr Gln Glu Val Glu Lys Leu
Asn Glu Ala Phe 515 520 525 Asp Ile Leu Leu Ala Phe Phe Ile Leu Ala
Cys Val Leu Ile Ile Phe 530 535 540 Leu Ile Tyr Lys Val Val Gln Phe
Lys Gln Lys Leu Lys Ala Ser Glu 545 550 555 560 Asn Ser Arg Glu Asn
Arg Leu Glu Tyr Tyr Ser Phe Tyr Gln Ser Ala 565 570 575 Arg Tyr Asn
Val Thr Ala Ser Ile Cys Asn Thr Ser Pro Asn Ser Leu 580 585 590 Glu
Ser Pro Gly Leu Glu Gln Ile Arg Leu His Lys Gln Ile Val Pro 595 600
605 Glu Asn Glu Ala Gln Val Ile Leu Phe Glu His Ser Ala Leu 610 615
620 <210> SEQ ID NO 24 <211> LENGTH: 31 <212>
TYPE: PRT <213> ORGANISM: Homo sapiens <400> SEQUENCE:
24 Met Cys Gly Leu Gln Phe Ser Leu Pro Cys Leu Arg Leu Phe Leu Val
1 5 10 15 Val Thr Cys Tyr Leu Leu Leu Leu Leu His Lys Glu Ile Leu
Gly 20 25 30 <210> SEQ ID NO 25 <211> LENGTH: 591
<212> TYPE: PRT <213> ORGANISM: Homo sapiens
<400> SEQUENCE: 25 Cys Ser Ser Val Cys Gln Leu Cys Thr Gly
Arg Gln Ile Asn Cys Arg 1 5 10 15 Asn Leu Gly Leu Ser Ser Ile Pro
Lys Asn Phe Pro Glu Ser Thr Val 20 25 30 Phe Leu Tyr Leu Thr Gly
Asn Asn Ile Ser Tyr Ile Asn Glu Ser Glu 35 40 45 Leu Thr Gly Leu
His Ser Leu Val Ala Leu Tyr Leu Asp Asn Ser Asn 50 55 60 Ile Leu
Tyr Val Tyr Pro Lys Ala Phe Val Gln Leu Arg His Leu Tyr 65 70 75 80
Phe Leu Phe Leu Asn Asn Asn Phe Ile Lys Arg Leu Asp Pro Gly Ile 85
90 95 Phe Lys Gly Leu Leu Asn Leu Arg Asn Leu Tyr Leu Gln Tyr Asn
Gln 100 105 110 Val Ser Phe Val Pro Arg Gly Val Phe Asn Asp Leu Val
Ser Val Gln 115 120 125 Tyr Leu Asn Leu Gln Arg Asn Arg Leu Thr Val
Leu Gly Ser Gly Thr 130 135 140 Phe Val Gly Met Val Ala Leu Arg Ile
Leu Asp Leu Ser Asn Asn Asn 145 150 155 160 Ile Leu Arg Ile Ser Glu
Ser Gly Phe Gln His Leu Glu Asn Leu Ala 165 170 175 Cys Leu Tyr Leu
Gly Ser Asn Asn Leu Thr Lys Val Pro Ser Asn Ala 180 185 190 Phe Glu
Val Leu Lys Ser Leu Arg Arg Leu Ser Leu Ser His Asn Pro 195 200 205
Ile Glu Ala Ile Gln Pro Phe Ala Phe Lys Gly Leu Ala Asn Leu Glu 210
215 220 Tyr Leu Leu Leu Lys Asn Ser Arg Ile Arg Asn Val Thr Arg Asp
Gly 225 230 235 240 Phe Ser Gly Ile Asn Asn Leu Lys His Leu Ile Leu
Ser His Asn Asp 245 250 255 Leu Glu Asn Leu Asn Ser Asp Thr Phe Ser
Leu Leu Lys Asn Leu Ile 260 265 270 Tyr Leu Lys Leu Asp Arg Asn Arg
Ile Ile Ser Ile Asp Asn Asp Thr 275 280 285 Phe Glu Asn Met Gly Ala
Ser Leu Lys Ile Leu Asn Leu Ser Phe Asn 290 295 300 Asn Leu Thr Ala
Leu His Pro Arg Val Leu Lys Pro Leu Ser Ser Leu 305 310 315 320 Ile
His Leu Gln Ala Asn Ser Asn Pro Trp Glu Cys Asn Cys Lys Leu 325 330
335 Leu Gly Leu Arg Asp Trp Leu Ala Ser Ser Ala Ile Thr Leu Asn Ile
340 345 350 Tyr Cys Gln Asn Pro Pro Ser Met Arg Gly Arg Ala Leu Arg
Tyr Ile 355 360 365 Asn Ile Thr Asn Cys Val Thr Ser Ser Ile Asn Val
Ser Arg Ala Trp 370 375 380 Ala Val Val Lys Ser Pro His Ile His His
Lys Thr Thr Ala Leu Met 385 390 395 400 Met Ala Trp His Lys Val Thr
Thr Asn Gly Ser Pro Leu Glu Asn Thr 405 410 415 Glu Thr Glu Asn Ile
Thr Phe Trp Glu Arg Ile Pro Thr Ser Pro Ala 420 425 430 Gly Arg Phe
Phe Gln Glu Asn Ala Phe Gly Asn Pro Leu Glu Thr Thr 435 440 445 Ala
Val Leu Pro Val Gln Ile Gln Leu Thr Thr Ser Val Thr Leu Asn 450 455
460 Leu Glu Lys Asn Ser Ala Leu Pro Asn Asp Ala Ala Ser Met Ser Gly
465 470 475 480 Lys Thr Ser Leu Ile Cys Thr Gln Glu Val Glu Lys Leu
Asn Glu Ala 485 490 495 Phe Asp Ile Leu Leu Ala Phe Phe Ile Leu Ala
Cys Val Leu Ile Ile 500 505 510 Phe Leu Ile Tyr Lys Val Val Gln Phe
Lys Gln Lys Leu Lys Ala Ser 515 520 525 Glu Asn Ser Arg Glu Asn Arg
Leu Glu Tyr Tyr Ser Phe Tyr Gln Ser 530 535 540 Ala Arg Tyr Asn Val
Thr Ala Ser Ile Cys Asn Thr Ser Pro Asn Ser 545 550 555 560 Leu Glu
Ser Pro Gly Leu Glu Gln Ile Arg Leu His Lys Gln Ile Val 565 570 575
Pro Glu Asn Glu Ala Gln Val Ile Leu Phe Glu His Ser Ala Leu 580 585
590 <210> SEQ ID NO 26 <211> LENGTH: 498 <212>
TYPE: PRT <213> ORGANISM: Homo sapiens <400> SEQUENCE:
26 Cys Ser Ser Val Cys Gln Leu Cys Thr Gly Arg Gln Ile Asn Cys Arg
1 5 10 15 Asn Leu Gly Leu Ser Ser Ile Pro Lys Asn Phe Pro Glu Ser
Thr Val 20 25 30 Phe Leu Tyr Leu Thr Gly Asn Asn Ile Ser Tyr Ile
Asn Glu Ser Glu 35 40 45 Leu Thr Gly Leu His Ser Leu Val Ala Leu
Tyr Leu Asp Asn Ser Asn 50 55 60 Ile Leu Tyr Val Tyr Pro Lys Ala
Phe Val Gln Leu Arg His Leu Tyr 65 70 75 80
Phe Leu Phe Leu Asn Asn Asn Phe Ile Lys Arg Leu Asp Pro Gly Ile 85
90 95 Phe Lys Gly Leu Leu Asn Leu Arg Asn Leu Tyr Leu Gln Tyr Asn
Gln 100 105 110 Val Ser Phe Val Pro Arg Gly Val Phe Asn Asp Leu Val
Ser Val Gln 115 120 125 Tyr Leu Asn Leu Gln Arg Asn Arg Leu Thr Val
Leu Gly Ser Gly Thr 130 135 140 Phe Val Gly Met Val Ala Leu Arg Ile
Leu Asp Leu Ser Asn Asn Asn 145 150 155 160 Ile Leu Arg Ile Ser Glu
Ser Gly Phe Gln His Leu Glu Asn Leu Ala 165 170 175 Cys Leu Tyr Leu
Gly Ser Asn Asn Leu Thr Lys Val Pro Ser Asn Ala 180 185 190 Phe Glu
Val Leu Lys Ser Leu Arg Arg Leu Ser Leu Ser His Asn Pro 195 200 205
Ile Glu Ala Ile Gln Pro Phe Ala Phe Lys Gly Leu Ala Asn Leu Glu 210
215 220 Tyr Leu Leu Leu Lys Asn Ser Arg Ile Arg Asn Val Thr Arg Asp
Gly 225 230 235 240 Phe Ser Gly Ile Asn Asn Leu Lys His Leu Ile Leu
Ser His Asn Asp 245 250 255 Leu Glu Asn Leu Asn Ser Asp Thr Phe Ser
Leu Leu Lys Asn Leu Ile 260 265 270 Tyr Leu Lys Leu Asp Arg Asn Arg
Ile Ile Ser Ile Asp Asn Asp Thr 275 280 285 Phe Glu Asn Met Gly Ala
Ser Leu Lys Ile Leu Asn Leu Ser Phe Asn 290 295 300 Asn Leu Thr Ala
Leu His Pro Arg Val Leu Lys Pro Leu Ser Ser Leu 305 310 315 320 Ile
His Leu Gln Ala Asn Ser Asn Pro Trp Glu Cys Asn Cys Lys Leu 325 330
335 Leu Gly Leu Arg Asp Trp Leu Ala Ser Ser Ala Ile Thr Leu Asn Ile
340 345 350 Tyr Cys Gln Asn Pro Pro Ser Met Arg Gly Arg Ala Leu Arg
Tyr Ile 355 360 365 Asn Ile Thr Asn Cys Val Thr Ser Ser Ile Asn Val
Ser Arg Ala Trp 370 375 380 Ala Val Val Lys Ser Pro His Ile His His
Lys Thr Thr Ala Leu Met 385 390 395 400 Met Ala Trp His Lys Val Thr
Thr Asn Gly Ser Pro Leu Glu Asn Thr 405 410 415 Glu Thr Glu Asn Ile
Thr Phe Trp Glu Arg Ile Pro Thr Ser Pro Ala 420 425 430 Gly Arg Phe
Phe Gln Glu Asn Ala Phe Gly Asn Pro Leu Glu Thr Thr 435 440 445 Ala
Val Leu Pro Val Gln Ile Gln Leu Thr Thr Ser Val Thr Leu Asn 450 455
460 Leu Glu Lys Asn Ser Ala Leu Pro Asn Asp Ala Ala Ser Met Ser Gly
465 470 475 480 Lys Thr Ser Leu Ile Cys Thr Gln Glu Val Glu Lys Leu
Asn Glu Ala 485 490 495 Phe Asp <210> SEQ ID NO 27
<211> LENGTH: 18 <212> TYPE: PRT <213> ORGANISM:
Homo sapiens <400> SEQUENCE: 27 Ile Leu Leu Ala Phe Phe Ile
Leu Ala Cys Val Leu Ile Ile Phe Leu 1 5 10 15 Ile Tyr <210>
SEQ ID NO 28 <211> LENGTH: 75 <212> TYPE: PRT
<213> ORGANISM: Homo sapiens <400> SEQUENCE: 28 Lys Val
Val Gln Phe Lys Gln Lys Leu Lys Ala Ser Glu Asn Ser Arg 1 5 10 15
Glu Asn Arg Leu Glu Tyr Tyr Ser Phe Tyr Gln Ser Ala Arg Tyr Asn 20
25 30 Val Thr Ala Ser Ile Cys Asn Thr Ser Pro Asn Ser Leu Glu Ser
Pro 35 40 45 Gly Leu Glu Gln Ile Arg Leu His Lys Gln Ile Val Pro
Glu Asn Glu 50 55 60 Ala Gln Val Ile Leu Phe Glu His Ser Ala Leu 65
70 75 <210> SEQ ID NO 29 <211> LENGTH: 1529 <212>
TYPE: PRT <213> ORGANISM: Homo sapiens <400> SEQUENCE:
29 Met Arg Gly Val Gly Trp Gln Met Leu Ser Leu Ser Leu Gly Leu Val
1 5 10 15 Leu Ala Ile Leu Asn Lys Val Ala Pro Gln Ala Cys Pro Ala
Gln Cys 20 25 30 Ser Cys Ser Gly Ser Thr Val Asp Cys His Gly Leu
Ala Leu Arg Ser 35 40 45 Val Pro Arg Asn Ile Pro Arg Asn Thr Glu
Arg Leu Asp Leu Asn Gly 50 55 60 Asn Asn Ile Thr Arg Ile Thr Lys
Thr Asp Phe Ala Gly Leu Arg His 65 70 75 80 Leu Arg Val Leu Gln Leu
Met Glu Asn Lys Ile Ser Thr Ile Glu Arg 85 90 95 Gly Ala Phe Gln
Asp Leu Lys Glu Leu Glu Arg Leu Arg Leu Asn Arg 100 105 110 Asn His
Leu Gln Leu Phe Pro Glu Leu Leu Phe Leu Gly Thr Ala Lys 115 120 125
Leu Tyr Arg Leu Asp Leu Ser Glu Asn Gln Ile Gln Ala Ile Pro Arg 130
135 140 Lys Ala Phe Arg Gly Ala Val Asp Ile Lys Asn Leu Gln Leu Asp
Tyr 145 150 155 160 Asn Gln Ile Ser Cys Ile Glu Asp Gly Ala Phe Arg
Ala Leu Arg Asp 165 170 175 Leu Glu Val Leu Thr Leu Asn Asn Asn Asn
Ile Thr Arg Leu Ser Val 180 185 190 Ala Ser Phe Asn His Met Pro Lys
Leu Arg Thr Phe Arg Leu His Ser 195 200 205 Asn Asn Leu Tyr Cys Asp
Cys His Leu Ala Trp Leu Ser Asp Trp Leu 210 215 220 Arg Gln Arg Pro
Arg Val Gly Leu Tyr Thr Gln Cys Met Gly Pro Ser 225 230 235 240 His
Leu Arg Gly His Asn Val Ala Glu Val Gln Lys Arg Glu Phe Val 245 250
255 Cys Ser Gly His Gln Ser Phe Met Ala Pro Ser Cys Ser Val Leu His
260 265 270 Cys Pro Ala Ala Cys Thr Cys Ser Asn Asn Ile Val Asp Cys
Arg Gly 275 280 285 Lys Gly Leu Thr Glu Ile Pro Thr Asn Leu Pro Glu
Thr Ile Thr Glu 290 295 300 Ile Arg Leu Glu Gln Asn Thr Ile Lys Val
Ile Pro Pro Gly Ala Phe 305 310 315 320 Ser Pro Tyr Lys Lys Leu Arg
Arg Ile Asp Leu Ser Asn Asn Gln Ile 325 330 335 Ser Glu Leu Ala Pro
Asp Ala Phe Gln Gly Leu Arg Ser Leu Asn Ser 340 345 350 Leu Val Leu
Tyr Gly Asn Lys Ile Thr Glu Leu Pro Lys Ser Leu Phe 355 360 365 Glu
Gly Leu Phe Ser Leu Gln Leu Leu Leu Leu Asn Ala Asn Lys Ile 370 375
380 Asn Cys Leu Arg Val Asp Ala Phe Gln Asp Leu His Asn Leu Asn Leu
385 390 395 400 Leu Ser Leu Tyr Asp Asn Lys Leu Gln Thr Ile Ala Lys
Gly Thr Phe 405 410 415 Ser Pro Leu Arg Ala Ile Gln Thr Met His Leu
Ala Gln Asn Pro Phe 420 425 430 Ile Cys Asp Cys His Leu Lys Trp Leu
Ala Asp Tyr Leu His Thr Asn 435 440 445 Pro Ile Glu Thr Ser Gly Ala
Arg Cys Thr Ser Pro Arg Arg Leu Ala 450 455 460 Asn Lys Arg Ile Gly
Gln Ile Lys Ser Lys Lys Phe Arg Cys Ser Ala 465 470 475 480 Lys Glu
Gln Tyr Phe Ile Pro Gly Thr Glu Asp Tyr Arg Ser Lys Leu 485 490 495
Ser Gly Asp Cys Phe Ala Asp Leu Ala Cys Pro Glu Lys Cys Arg Cys 500
505 510 Glu Gly Thr Thr Val Asp Cys Ser Asn Gln Lys Leu Asn Lys Ile
Pro 515 520 525 Glu His Ile Pro Gln Tyr Thr Ala Glu Leu Arg Leu Asn
Asn Asn Glu 530 535 540 Phe Thr Val Leu Glu Ala Thr Gly Ile Phe Lys
Lys Leu Pro Gln Leu 545 550 555 560 Arg Lys Ile Asn Phe Ser Asn Asn
Lys Ile Thr Asp Ile Glu Glu Gly 565 570 575 Ala Phe Glu Gly Ala Ser
Gly Val Asn Glu Ile Leu Leu Thr Ser Asn 580 585 590 Arg Leu Glu Asn
Val Gln His Lys Met Phe Lys Gly Leu Glu Ser Leu 595 600 605 Lys Thr
Leu Met Leu Arg Ser Asn Arg Ile Thr Cys Val Gly Asn Asp 610 615 620
Ser Phe Ile Gly Leu Ser Ser Val Arg Leu Leu Ser Leu Tyr Asp Asn 625
630 635 640 Gln Ile Thr Thr Val Ala Pro Gly Ala Phe Asp Thr Leu His
Ser Leu 645 650 655 Ser Thr Leu Asn Leu Leu Ala Asn Pro Phe Asn Cys
Asn Cys Tyr Leu 660 665 670 Ala Trp Leu Gly Glu Trp Leu Arg Lys Lys
Arg Ile Val Thr Gly Asn
675 680 685 Pro Arg Cys Gln Lys Pro Tyr Phe Leu Lys Glu Ile Pro Ile
Gln Asp 690 695 700 Val Ala Ile Gln Asp Phe Thr Cys Asp Asp Gly Asn
Asp Asp Asn Ser 705 710 715 720 Cys Ser Pro Leu Ser Arg Cys Pro Thr
Glu Cys Thr Cys Leu Asp Thr 725 730 735 Val Val Arg Cys Ser Asn Lys
Gly Leu Lys Val Leu Pro Lys Gly Ile 740 745 750 Pro Arg Asp Val Thr
Glu Leu Tyr Leu Asp Gly Asn Gln Phe Thr Leu 755 760 765 Val Pro Lys
Glu Leu Ser Asn Tyr Lys His Leu Thr Leu Ile Asp Leu 770 775 780 Ser
Asn Asn Arg Ile Ser Thr Leu Ser Asn Gln Ser Phe Ser Asn Met 785 790
795 800 Thr Gln Leu Leu Thr Leu Ile Leu Ser Tyr Asn Arg Leu Arg Cys
Ile 805 810 815 Pro Pro Arg Thr Phe Asp Gly Leu Lys Ser Leu Arg Leu
Leu Ser Leu 820 825 830 His Gly Asn Asp Ile Ser Val Val Pro Glu Gly
Ala Phe Asn Asp Leu 835 840 845 Ser Ala Leu Ser His Leu Ala Ile Gly
Ala Asn Pro Leu Tyr Cys Asp 850 855 860 Cys Asn Met Gln Trp Leu Ser
Asp Trp Val Lys Ser Glu Tyr Lys Glu 865 870 875 880 Pro Gly Ile Ala
Arg Cys Ala Gly Pro Gly Glu Met Ala Asp Lys Leu 885 890 895 Leu Leu
Thr Thr Pro Ser Lys Lys Phe Thr Cys Gln Gly Pro Val Asp 900 905 910
Val Asn Ile Leu Ala Lys Cys Asn Pro Cys Leu Ser Asn Pro Cys Lys 915
920 925 Asn Asp Gly Thr Cys Asn Ser Asp Pro Val Asp Phe Tyr Arg Cys
Thr 930 935 940 Cys Pro Tyr Gly Phe Lys Gly Gln Asp Cys Asp Val Pro
Ile His Ala 945 950 955 960 Cys Ile Ser Asn Pro Cys Lys His Gly Gly
Thr Cys His Leu Lys Glu 965 970 975 Gly Glu Glu Asp Gly Phe Trp Cys
Ile Cys Ala Asp Gly Phe Glu Gly 980 985 990 Glu Asn Cys Glu Val Asn
Val Asp Asp Cys Glu Asp Asn Asp Cys Glu 995 1000 1005 Asn Asn Ser
Thr Cys Val Asp Gly Ile Asn Asn Tyr Thr Cys Leu 1010 1015 1020 Cys
Pro Pro Glu Tyr Thr Gly Glu Leu Cys Glu Glu Lys Leu Asp 1025 1030
1035 Phe Cys Ala Gln Asp Leu Asn Pro Cys Gln His Asp Ser Lys Cys
1040 1045 1050 Ile Leu Thr Pro Lys Gly Phe Lys Cys Asp Cys Thr Pro
Gly Tyr 1055 1060 1065 Val Gly Glu His Cys Asp Ile Asp Phe Asp Asp
Cys Gln Asp Asn 1070 1075 1080 Lys Cys Lys Asn Gly Ala His Cys Thr
Asp Ala Val Asn Gly Tyr 1085 1090 1095 Thr Cys Ile Cys Pro Glu Gly
Tyr Ser Gly Leu Phe Cys Glu Phe 1100 1105 1110 Ser Pro Pro Met Val
Leu Pro Arg Thr Ser Pro Cys Asp Asn Phe 1115 1120 1125 Asp Cys Gln
Asn Gly Ala Gln Cys Ile Val Arg Ile Asn Glu Pro 1130 1135 1140 Ile
Cys Gln Cys Leu Pro Gly Tyr Gln Gly Glu Lys Cys Glu Lys 1145 1150
1155 Leu Val Ser Val Asn Phe Ile Asn Lys Glu Ser Tyr Leu Gln Ile
1160 1165 1170 Pro Ser Ala Lys Val Arg Pro Gln Thr Asn Ile Thr Leu
Gln Ile 1175 1180 1185 Ala Thr Asp Glu Asp Ser Gly Ile Leu Leu Tyr
Lys Gly Asp Lys 1190 1195 1200 Asp His Ile Ala Val Glu Leu Tyr Arg
Gly Arg Val Arg Ala Ser 1205 1210 1215 Tyr Asp Thr Gly Ser His Pro
Ala Ser Ala Ile Tyr Ser Val Glu 1220 1225 1230 Thr Ile Asn Asp Gly
Asn Phe His Ile Val Glu Leu Leu Ala Leu 1235 1240 1245 Asp Gln Ser
Leu Ser Leu Ser Val Asp Gly Gly Asn Pro Lys Ile 1250 1255 1260 Ile
Thr Asn Leu Ser Lys Gln Ser Thr Leu Asn Phe Asp Ser Pro 1265 1270
1275 Leu Tyr Val Gly Gly Met Pro Gly Lys Ser Asn Val Ala Ser Leu
1280 1285 1290 Arg Gln Ala Pro Gly Gln Asn Gly Thr Ser Phe His Gly
Cys Ile 1295 1300 1305 Arg Asn Leu Tyr Ile Asn Ser Glu Leu Gln Asp
Phe Gln Lys Val 1310 1315 1320 Pro Met Gln Thr Gly Ile Leu Pro Gly
Cys Glu Pro Cys His Lys 1325 1330 1335 Lys Val Cys Ala His Gly Thr
Cys Gln Pro Ser Ser Gln Ala Gly 1340 1345 1350 Phe Thr Cys Glu Cys
Gln Glu Gly Trp Met Gly Pro Leu Cys Asp 1355 1360 1365 Gln Arg Thr
Asn Asp Pro Cys Leu Gly Asn Lys Cys Val His Gly 1370 1375 1380 Thr
Cys Leu Pro Ile Asn Ala Phe Ser Tyr Ser Cys Lys Cys Leu 1385 1390
1395 Glu Gly His Gly Gly Val Leu Cys Asp Glu Glu Glu Asp Leu Phe
1400 1405 1410 Asn Pro Cys Gln Ala Ile Lys Cys Lys His Gly Lys Cys
Arg Leu 1415 1420 1425 Ser Gly Leu Gly Gln Pro Tyr Cys Glu Cys Ser
Ser Gly Tyr Thr 1430 1435 1440 Gly Asp Ser Cys Asp Arg Glu Ile Ser
Cys Arg Gly Glu Arg Ile 1445 1450 1455 Arg Asp Tyr Tyr Gln Lys Gln
Gln Gly Tyr Ala Ala Cys Gln Thr 1460 1465 1470 Thr Lys Lys Val Ser
Arg Leu Glu Cys Arg Gly Gly Cys Ala Gly 1475 1480 1485 Gly Gln Cys
Cys Gly Pro Leu Arg Ser Lys Arg Arg Lys Tyr Ser 1490 1495 1500 Phe
Glu Cys Thr Asp Gly Ser Ser Phe Val Asp Glu Val Glu Lys 1505 1510
1515 Val Val Lys Cys Gly Cys Thr Arg Cys Val Ser 1520 1525
<210> SEQ ID NO 30 <211> LENGTH: 4900 <212> TYPE:
DNA <213> ORGANISM: Homo sapiens <400> SEQUENCE: 30
cagagcaggg tggagagggc ggtgggaggc gtgtgcctga gtgggctcta ctgccttgtt
60 ccatattatt ttgtgcacat tttccctggc actctgggtt gctagccccg
ccgggcactg 120 ggcctcagac actgcgcggt tccctcggag cagcaagcta
aagaaagccc ccagtgccgg 180 cgaggaagga ggcggcgggg aaagatgcgc
ggcgttggct ggcagatgct gtccctgtcg 240 ctggggttag tgctggcgat
cctgaacaag gtggcaccgc aggcgtgccc ggcgcagtgc 300 tcttgctcgg
gcagcacagt ggactgtcac gggctggcgc tgcgcagcgt gcccaggaat 360
atcccccgca acaccgagag actggattta aatggaaata acatcacaag aattacgaag
420 acagattttg ctggtcttag acatctaaga gttcttcagc ttatggagaa
taagattagc 480 accattgaaa gaggagcatt ccaggatctt aaagaactag
agagactgcg tttaaacaga 540 aatcaccttc agctgtttcc tgagttgctg
tttcttggga ctgcgaagct atacaggctt 600 gatctcagtg aaaaccaaat
tcaggcaatc ccaaggaaag ctttccgtgg ggcagttgac 660 ataaaaaatt
tgcaactgga ttacaaccag atcagctgta ttgaagatgg ggcattcagg 720
gctctccggg acctggaagt gctcactctc aacaataaca acattactag actttctgtg
780 gcaagtttca accatatgcc taaacttagg acttttcgac tgcattcaaa
caacctgtat 840 tgtgactgcc acctggcctg gctctccgac tggcttcgcc
aaaggcctcg ggttggtctg 900 tacactcagt gtatgggccc ctcccacctg
agaggccata atgtagccga ggttcaaaaa 960 cgagaatttg tctgcagtgg
tcaccagtca tttatggctc cttcttgtag tgttttgcac 1020 tgccctgccg
cctgtacctg tagcaacaat atcgtagact gtcgtgggaa aggtctcact 1080
gagatcccca caaatcttcc agagaccatc acagaaatac gtttggaaca gaacacaatc
1140 aaagtcatcc ctcctggagc tttctcacca tataaaaagc ttagacgaat
tgacctgagc 1200 aataatcaga tctctgaact tgcaccagat gctttccaag
gactacgctc tctgaattca 1260 cttgtcctct atggaaataa aatcacagaa
ctccccaaaa gtttatttga aggactgttt 1320 tccttacagc tcctattatt
gaatgccaac aagataaact gccttcgggt agatgctttt 1380 caggatctcc
acaacttgaa ccttctctcc ctatatgaca acaagcttca gaccatcgcc 1440
aaggggacct tttcacctct tcgggccatt caaactatgc atttggccca gaaccccttt
1500 atttgtgact gccatctcaa gtggctagcg gattatctcc ataccaaccc
gattgagacc 1560 agtggtgccc gttgcaccag cccccgccgc ctggcaaaca
aaagaattgg acagatcaaa 1620 agcaagaaat tccgttgttc agctaaagaa
cagtatttca ttccaggtac agaagattat 1680 cgatcaaaat taagtggaga
ctgctttgcg gatctggctt gccctgaaaa gtgtcgctgt 1740 gaaggaacca
cagtagattg ctctaatcaa aagctcaaca aaatcccgga gcacattccc 1800
cagtacactg cagagttgcg tctcaataat aatgaattta ccgtgttgga agccacagga
1860 atctttaaga aacttcctca attacgtaaa ataaacttta gcaacaataa
gatcacagat 1920 attgaggagg gagcatttga aggagcatct ggtgtaaatg
aaatacttct tacgagtaat 1980 cgtttggaaa atgtgcagca taagatgttc
aagggattgg aaagcctcaa aactttgatg 2040 ttgagaagca atcgaataac
ctgtgtgggg aatgacagtt tcataggact cagttctgtg 2100 cgtttgcttt
ctttgtatga taatcaaatt actacagttg caccaggggc atttgatact 2160
ctccattctt tatctactct aaacctcttg gccaatcctt ttaactgtaa ctgctacctg
2220 gcttggttgg gagagtggct gagaaagaag agaattgtca cgggaaatcc
tagatgtcaa 2280
aaaccatact tcctgaaaga aatacccatc caggatgtgg ccattcagga cttcacttgt
2340 gatgacggaa atgatgacaa tagttgctcc ccactttctc gctgtcctac
tgaatgtact 2400 tgcttggata cagtcgtccg atgtagcaac aagggtttga
aggtcttgcc gaaaggtatt 2460 ccaagagatg tcacagagtt gtatctggat
ggaaaccaat ttacactggt tcccaaggaa 2520 ctctccaact acaaacattt
aacacttata gacttaagta acaacagaat aagcacgctt 2580 tctaatcaga
gcttcagcaa catgacccag ctcctcacct taattcttag ttacaaccgt 2640
ctgagatgta ttcctcctcg cacctttgat ggattaaagt ctcttcgatt actttctcta
2700 catggaaatg acatttctgt tgtgcctgaa ggtgctttca atgatctttc
tgcattatca 2760 catctagcaa ttggagccaa ccctctttac tgtgattgta
acatgcagtg gttatccgac 2820 tgggtgaagt cggaatataa ggagcctgga
attgctcgtt gtgctggtcc tggagaaatg 2880 gcagataaac ttttactcac
aactccctcc aaaaaattta cctgtcaagg tcctgtggat 2940 gtcaatattc
tagctaagtg taacccctgc ctatcaaatc cgtgtaaaaa tgatggcaca 3000
tgtaatagtg atccagttga cttttaccga tgcacctgtc catatggttt caaggggcag
3060 gactgtgatg tcccaattca tgcctgcatc agtaacccat gtaaacatgg
aggaacttgc 3120 cacttaaagg aaggagaaga agatggattc tggtgtattt
gtgctgatgg atttgaagga 3180 gaaaattgtg aagtcaacgt tgatgattgt
gaagataatg actgtgaaaa taattctaca 3240 tgtgtcgatg gcattaataa
ctacacatgc ctttgcccac ctgagtatac aggtgagttg 3300 tgtgaggaga
agctggactt ctgtgcccag gacctgaacc cctgccagca cgattcaaag 3360
tgcatcctaa ctccaaaggg attcaaatgt gactgcacac cagggtacgt aggtgaacac
3420 tgcgacatcg attttgacga ctgccaagac aacaagtgta aaaacggagc
ccactgcaca 3480 gatgcagtga acggctatac gtgcatatgc cccgaaggtt
acagtggctt gttctgtgag 3540 ttttctccac ccatggtcct ccctcgtacc
agcccctgtg ataattttga ttgtcagaat 3600 ggagctcagt gtatcgtcag
aataaatgag ccaatatgtc agtgtttgcc tggctatcag 3660 ggagaaaagt
gtgaaaaatt ggttagtgtg aattttataa acaaagagtc ttatcttcag 3720
attccttcag ccaaggttcg gcctcagacg aacataacac ttcagattgc cacagatgaa
3780 gacagcggaa tcctcctgta taagggtgac aaagaccata tcgcggtaga
actctatcgg 3840 gggcgtgttc gtgccagcta tgacaccggc tctcatccag
cttctgccat ttacagtgtg 3900 gagacaatca atgatggaaa cttccacatt
gtggaactac ttgccttgga tcagagtctc 3960 tctttgtccg tggatggtgg
gaaccccaaa atcatcacta acttgtcaaa gcagtccact 4020 ctgaattttg
actctccact ctatgtagga ggcatgccag ggaagagtaa cgtggcatct 4080
ctgcgccagg cccctgggca gaacggaacc agcttccacg gctgcatccg gaacctttac
4140 atcaacagtg agctgcagga cttccagaag gtgccgatgc aaacaggcat
tttgcctggc 4200 tgtgagccat gccacaagaa ggtgtgtgcc catggcacat
gccagcccag cagccaggca 4260 ggcttcacct gcgagtgcca ggaaggatgg
atggggcccc tctgtgacca acggaccaat 4320 gacccttgcc ttggaaataa
atgcgtacat ggcacctgct tgcccatcaa tgcgttctcc 4380 tacagctgta
agtgcttgga gggccatgga ggtgtcctct gtgatgaaga ggaggatctg 4440
tttaacccat gccaggcgat caagtgcaag cacgggaagt gcaggctttc aggtctgggg
4500 cagccctact gtgaatgcag cagtggatac acgggggaca gctgtgatcg
agaaatctct 4560 tgtcgagggg aaaggataag agattattac caaaagcagc
agggctatgc tgcttgccaa 4620 acaaccaaga aggtgtcccg attagagtgc
agaggtgggt gtgcaggagg gcagtgctgt 4680 ggaccgctga ggagcaagcg
gcggaaatac tctttcgaat gcactgacgg ctcctccttt 4740 gtggacgagg
ttgagaaagt ggtgaagtgc ggctgtacga ggtgtgtgtc ctaaacacac 4800
tcccggcagc tctgtctttg gaaaaggttg tatacttctt gaccatgtgg gactaatgaa
4860 tgcttcatag tggaaatatt tgaaatatat tgtaaaatac 4900 <210>
SEQ ID NO 31 <211> LENGTH: 3510 <212> TYPE: DNA
<213> ORGANISM: Homo sapiens <400> SEQUENCE: 31
gcagctctgg gggagctcgg agctcccgat cacggcttct tgggggtagc tacggctggg
60 tgtgtagaac ggggccgggg ctggggctgg gtcccctagt ggagacccaa
gtgcgagagg 120 caagaactct gcagcttcct gccttctggg tcagttcctt
attcaagtct gcagccggct 180 cccagggaga tctcggtgga acttcagaaa
cgctgggcag tctgcctttc aaccatgccc 240 ctgtccctgg gagccgagat
gtgggggcct gaggcctggc tgctgctgct gctactgctg 300 gcatcattta
caggccggtg ccccgcgggt gagctggaga cctcagacgt ggtaactgtg 360
gtgctgggcc aggacgcaaa actgccctgc ttctaccgag gggactccgg cgagcaagtg
420 gggcaagtgg catgggctcg ggtggacgcg ggcgaaggcg cccaggaact
agcgctactg 480 cactccaaat acgggcttca tgtgagcccg gcttacgagg
gccgcgtgga gcagccgccg 540 cccccacgca accccctgga cggctcagtg
ctcctgcgca acgcagtgca ggcggatgag 600 ggcgagtacg agtgccgggt
cagcaccttc cccgccggca gcttccaggc gcggctgcgg 660 ctccgagtgc
tggtgcctcc cctgccctca ctgaatcctg gtccagcact agaagagggc 720
cagggcctga ccctggcagc ctcctgcaca gctgagggca gcccagcccc cagcgtgacc
780 tgggacacgg aggtcaaagg cacaacgtcc agccgttcct tcaagcactc
ccgctctgct 840 gccgtcacct cagagttcca cttggtgcct agccgcagca
tgaatgggca gccactgact 900 tgtgtggtgt cccatcctgg cctgctccag
gaccaaagga tcacccacat cctccacgtg 960 tccttccttg ctgaggcctc
tgtgaggggc cttgaagacc aaaatctgtg gcacattggc 1020 agagaaggag
ctatgctcaa gtgcctgagt gaagggcagc cccctccctc atacaactgg 1080
acacggctgg atgggcctct gcccagtggg gtacgagtgg atggggacac tttgggcttt
1140 cccccactga ccactgagca cagcggcatc tacgtctgcc atgtcagcaa
tgagttctcc 1200 tcaagggatt ctcaggtcac tgtggatgtt cttgaccccc
aggaagactc tgggaagcag 1260 gtggacctag tgtcagcctc ggtggtggtg
gtgggtgtga tcgccgcact cttgttctgc 1320 cttctggtgg tggtggtggt
gctcatgtcc cgataccatc ggcgcaaggc ccagcagatg 1380 acccagaaat
atgaggagga gctgaccctg accagggaga actccatccg gaggctgcat 1440
tcccatcaca cggaccccag gagccagccg gaggagagtg tagggctgag agccgagggc
1500 caccctgata gtctcaagga caacagtagc tgctctgtga tgagtgaaga
gcccgagggc 1560 cgcagttact ccacgctgac cacggtgagg gagatagaaa
cacagactga actgctgtct 1620 ccaggctctg ggcgggccga ggaggaggaa
gatcaggatg aaggcatcaa acaggccatg 1680 aaccattttg ttcaggagaa
tgggacccta cgggccaagc ccacgggcaa tggcatctac 1740 atcaatgggc
ggggacacct ggtctgaccc aggcctgcct cccttcccta ggcctggctc 1800
cttctgttga catgggagat tttagctcat cttgggggcc tccttaaaca cccccatttc
1860 ttgcggaaga tgctccccat cccactgact gcttgacctt tacctccaac
ccttctgttc 1920 atcgggaggg ctccaccaat tgagtctctc ccaccatgca
tgcaggtcac tgtgtgtgtg 1980 catgtgtgcc tgtgtgagtg ttgactgact
gtgtgtgtgt ggaggggtga ctgtccgtgg 2040 aggggtgact gtgtccgtgg
tgtgtattat gctgtcatat cagagtcaag tgaactgtgg 2100 tgtatgtgcc
acgggatttg agtggttgcg tgggcaacac tgtcagggtt tggcgtgtgt 2160
gtcatgtggc tgtgtgtgac ctctgcctga aaaagcaggt attttctcag accccagagc
2220 agtattaatg atgcagaggt tggaggagag aggtggagac tgtggctcag
acccaggtgt 2280 gcgggcatag ctggagctgg aatctgcctc cggtgtgagg
gaacctgtct cctaccactt 2340 cggagccatg ggggcaagtg tgaagcagcc
agtccctggg tcagccagag gcttgaactg 2400 ttacagaagc cctctgccct
ctggtggcct ctgggcctgc tgcatgtaca tattttctgt 2460 aaatatacat
gcgccgggag cttcttgcag gaatactgct ccgaatcact tttaattttt 2520
ttcttttttt tttcttgccc tttccattag ttgtattttt tatttatttt tatttttatt
2580 tttttttaga gatggagtct cactatgttg ctcaggctgg ccttgaactc
ctgggctcaa 2640 gcaatcctcc tgcctcagcc tccctagtag ctgggacttt
aagtgtacac cactgtgcct 2700 gctttgaatc ctttacgaag agaaaaaaaa
aattaaagaa agcctttaga tttatccaat 2760 gtttactact gggattgctt
aaagtgaggc ccctccaaca ccagggggtt aattcctgtg 2820 attgtgaaag
gggctacttc caaggcatct tcatgcaggc agccccttgg gagggcacct 2880
gagagctggt agagtctgaa attagggatg tgagcctcgt ggttactgag taaggtaaaa
2940 ttgcatccac cattgtttgt gataccttag ggaattgctt ggacctggtg
acaagggctc 3000 ctgttcaata gtggtgttgg ggagagagag agcagtgatt
atagaccgag agagtaggag 3060 ttgaggtgag gtgaaggagg tgctgggggt
gagaatgtcg cctttccccc tgggttttgg 3120 atcactaatt caaggctctt
ctggatgttt ctctgggttg gggctggagt tcaatgaggt 3180 ttatttttag
ctggcccacc cagatacact cagccagaat acctagattt agtacccaaa 3240
ctcttcttag tctgaaatct gctggatttc tggcctaagg gagaggctcc catccttcgt
3300 tccccagcca gcctaggact tcgaatgtgg agcctgaaga tctaagatcc
taacatgtac 3360 attttatgta aatatgtgca tatttgtaca taaaatgata
ttctgttttt aaataaacag 3420 acaaaacttg aaaaaaaaaa aaaaaaaaaa
aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa 3480 aaaaaaaaaa aaaaaaaaaa
aaaaaaaaaa 3510 <210> SEQ ID NO 32 <211> LENGTH: 1530
<212> TYPE: DNA <213> ORGANISM: Homo sapiens
<400> SEQUENCE: 32 atgcccctgt ccctgggagc cgagatgtgg
gggcctgagg cctggctgct gctgctgcta 60 ctgctggcat catttacagg
ccggtgcccc gcgggtgagc tggagacctc agacgtggta 120 actgtggtgc
tgggccagga cgcaaaactg ccctgcttct accgagggga ctccggcgag 180
caagtggggc aagtggcatg ggctcgggtg gacgcgggcg aaggcgccca ggaactagcg
240 ctactgcact ccaaatacgg gcttcatgtg agcccggctt acgagggccg
cgtggagcag 300 ccgccgcccc cacgcaaccc cctggacggc tcagtgctcc
tgcgcaacgc agtgcaggcg 360 gatgagggcg agtacgagtg ccgggtcagc
accttccccg ccggcagctt ccaggcgcgg 420 ctgcggctcc gagtgctggt
gcctcccctg ccctcactga atcctggtcc agcactagaa 480 gagggccagg
gcctgaccct ggcagcctcc tgcacagctg agggcagccc agcccccagc 540
gtgacctggg acacggaggt caaaggcaca acgtccagcc gttccttcaa gcactcccgc
600 tctgctgccg tcacctcaga gttccacttg gtgcctagcc gcagcatgaa
tgggcagcca 660 ctgacttgtg tggtgtccca tcctggcctg ctccaggacc
aaaggatcac ccacatcctc 720 cacgtgtcct tccttgctga ggcctctgtg
aggggccttg aagaccaaaa tctgtggcac 780 attggcagag aaggagctat
gctcaagtgc ctgagtgaag ggcagccccc tccctcatac 840 aactggacac
ggctggatgg gcctctgccc agtggggtac gagtggatgg ggacactttg 900
ggctttcccc cactgaccac tgagcacagc ggcatctacg tctgccatgt cagcaatgag
960 ttctcctcaa gggattctca ggtcactgtg gatgttcttg acccccagga
agactctggg 1020 aagcaggtgg acctagtgtc agcctcggtg gtggtggtgg
gtgtgatcgc cgcactcttg 1080 ttctgccttc tggtggtggt ggtggtgctc
atgtcccgat accatcggcg caaggcccag 1140 cagatgaccc agaaatatga
ggaggagctg accctgacca gggagaactc catccggagg 1200 ctgcattccc
atcacacgga ccccaggagc cagccggagg agagtgtagg gctgagagcc 1260
gagggccacc ctgatagtct caaggacaac agtagctgct ctgtgatgag tgaagagccc
1320 gagggccgca gttactccac gctgaccacg gtgagggaga tagaaacaca
gactgaactg 1380 ctgtctccag gctctgggcg ggccgaggag gaggaagatc
aggatgaagg catcaaacag 1440 gccatgaacc attttgttca ggagaatggg
accctacggg ccaagcccac gggcaatggc 1500 atctacatca atgggcgggg
acacctggtc 1530 <210> SEQ ID NO 33 <211> LENGTH: 510
<212> TYPE: PRT <213> ORGANISM: Homo sapiens
<400> SEQUENCE: 33 Met Pro Leu Ser Leu Gly Ala Glu Met Trp
Gly Pro Glu Ala Trp Leu 1 5 10 15 Leu Leu Leu Leu Leu Leu Ala Ser
Phe Thr Gly Arg Cys Pro Ala Gly 20 25 30 Glu Leu Glu Thr Ser Asp
Val Val Thr Val Val Leu Gly Gln Asp Ala 35 40 45 Lys Leu Pro Cys
Phe Tyr Arg Gly Asp Ser Gly Glu Gln Val Gly Gln 50 55 60 Val Ala
Trp Ala Arg Val Asp Ala Gly Glu Gly Ala Gln Glu Leu Ala 65 70 75 80
Leu Leu His Ser Lys Tyr Gly Leu His Val Ser Pro Ala Tyr Glu Gly 85
90 95 Arg Val Glu Gln Pro Pro Pro Pro Arg Asn Pro Leu Asp Gly Ser
Val 100 105 110 Leu Leu Arg Asn Ala Val Gln Ala Asp Glu Gly Glu Tyr
Glu Cys Arg 115 120 125 Val Ser Thr Phe Pro Ala Gly Ser Phe Gln Ala
Arg Leu Arg Leu Arg 130 135 140 Val Leu Val Pro Pro Leu Pro Ser Leu
Asn Pro Gly Pro Ala Leu Glu 145 150 155 160 Glu Gly Gln Gly Leu Thr
Leu Ala Ala Ser Cys Thr Ala Glu Gly Ser 165 170 175 Pro Ala Pro Ser
Val Thr Trp Asp Thr Glu Val Lys Gly Thr Thr Ser 180 185 190 Ser Arg
Ser Phe Lys His Ser Arg Ser Ala Ala Val Thr Ser Glu Phe 195 200 205
His Leu Val Pro Ser Arg Ser Met Asn Gly Gln Pro Leu Thr Cys Val 210
215 220 Val Ser His Pro Gly Leu Leu Gln Asp Gln Arg Ile Thr His Ile
Leu 225 230 235 240 His Val Ser Phe Leu Ala Glu Ala Ser Val Arg Gly
Leu Glu Asp Gln 245 250 255 Asn Leu Trp His Ile Gly Arg Glu Gly Ala
Met Leu Lys Cys Leu Ser 260 265 270 Glu Gly Gln Pro Pro Pro Ser Tyr
Asn Trp Thr Arg Leu Asp Gly Pro 275 280 285 Leu Pro Ser Gly Val Arg
Val Asp Gly Asp Thr Leu Gly Phe Pro Pro 290 295 300 Leu Thr Thr Glu
His Ser Gly Ile Tyr Val Cys His Val Ser Asn Glu 305 310 315 320 Phe
Ser Ser Arg Asp Ser Gln Val Thr Val Asp Val Leu Asp Pro Gln 325 330
335 Glu Asp Ser Gly Lys Gln Val Asp Leu Val Ser Ala Ser Val Val Val
340 345 350 Val Gly Val Ile Ala Ala Leu Leu Phe Cys Leu Leu Val Val
Val Val 355 360 365 Val Leu Met Ser Arg Tyr His Arg Arg Lys Ala Gln
Gln Met Thr Gln 370 375 380 Lys Tyr Glu Glu Glu Leu Thr Leu Thr Arg
Glu Asn Ser Ile Arg Arg 385 390 395 400 Leu His Ser His His Thr Asp
Pro Arg Ser Gln Pro Glu Glu Ser Val 405 410 415 Gly Leu Arg Ala Glu
Gly His Pro Asp Ser Leu Lys Asp Asn Ser Ser 420 425 430 Cys Ser Val
Met Ser Glu Glu Pro Glu Gly Arg Ser Tyr Ser Thr Leu 435 440 445 Thr
Thr Val Arg Glu Ile Glu Thr Gln Thr Glu Leu Leu Ser Pro Gly 450 455
460 Ser Gly Arg Ala Glu Glu Glu Glu Asp Gln Asp Glu Gly Ile Lys Gln
465 470 475 480 Ala Met Asn His Phe Val Gln Glu Asn Gly Thr Leu Arg
Ala Lys Pro 485 490 495 Thr Gly Asn Gly Ile Tyr Ile Asn Gly Arg Gly
His Leu Val 500 505 510 <210> SEQ ID NO 34 <211>
LENGTH: 31 <212> TYPE: PRT <213> ORGANISM: Homo sapiens
<400> SEQUENCE: 34 Met Pro Leu Ser Leu Gly Ala Glu Met Trp
Gly Pro Glu Ala Trp Leu 1 5 10 15 Leu Leu Leu Leu Leu Leu Ala Ser
Phe Thr Gly Arg Cys Pro Ala 20 25 30 <210> SEQ ID NO 35
<211> LENGTH: 479 <212> TYPE: PRT <213> ORGANISM:
Homo sapiens <400> SEQUENCE: 35 Gly Glu Leu Glu Thr Ser Asp
Val Val Thr Val Val Leu Gly Gln Asp 1 5 10 15 Ala Lys Leu Pro Cys
Phe Tyr Arg Gly Asp Ser Gly Glu Gln Val Gly 20 25 30 Gln Val Ala
Trp Ala Arg Val Asp Ala Gly Glu Gly Ala Gln Glu Leu 35 40 45 Ala
Leu Leu His Ser Lys Tyr Gly Leu His Val Ser Pro Ala Tyr Glu 50 55
60 Gly Arg Val Glu Gln Pro Pro Pro Pro Arg Asn Pro Leu Asp Gly Ser
65 70 75 80 Val Leu Leu Arg Asn Ala Val Gln Ala Asp Glu Gly Glu Tyr
Glu Cys 85 90 95 Arg Val Ser Thr Phe Pro Ala Gly Ser Phe Gln Ala
Arg Leu Arg Leu 100 105 110 Arg Val Leu Val Pro Pro Leu Pro Ser Leu
Asn Pro Gly Pro Ala Leu 115 120 125 Glu Glu Gly Gln Gly Leu Thr Leu
Ala Ala Ser Cys Thr Ala Glu Gly 130 135 140 Ser Pro Ala Pro Ser Val
Thr Trp Asp Thr Glu Val Lys Gly Thr Thr 145 150 155 160 Ser Ser Arg
Ser Phe Lys His Ser Arg Ser Ala Ala Val Thr Ser Glu 165 170 175 Phe
His Leu Val Pro Ser Arg Ser Met Asn Gly Gln Pro Leu Thr Cys 180 185
190 Val Val Ser His Pro Gly Leu Leu Gln Asp Gln Arg Ile Thr His Ile
195 200 205 Leu His Val Ser Phe Leu Ala Glu Ala Ser Val Arg Gly Leu
Glu Asp 210 215 220 Gln Asn Leu Trp His Ile Gly Arg Glu Gly Ala Met
Leu Lys Cys Leu 225 230 235 240 Ser Glu Gly Gln Pro Pro Pro Ser Tyr
Asn Trp Thr Arg Leu Asp Gly 245 250 255 Pro Leu Pro Ser Gly Val Arg
Val Asp Gly Asp Thr Leu Gly Phe Pro 260 265 270 Pro Leu Thr Thr Glu
His Ser Gly Ile Tyr Val Cys His Val Ser Asn 275 280 285 Glu Phe Ser
Ser Arg Asp Ser Gln Val Thr Val Asp Val Leu Asp Pro 290 295 300 Gln
Glu Asp Ser Gly Lys Gln Val Asp Leu Val Ser Ala Ser Val Val 305 310
315 320 Val Val Gly Val Ile Ala Ala Leu Leu Phe Cys Leu Leu Val Val
Val 325 330 335 Val Val Leu Met Ser Arg Tyr His Arg Arg Lys Ala Gln
Gln Met Thr 340 345 350 Gln Lys Tyr Glu Glu Glu Leu Thr Leu Thr Arg
Glu Asn Ser Ile Arg 355 360 365 Arg Leu His Ser His His Thr Asp Pro
Arg Ser Gln Pro Glu Glu Ser 370 375 380 Val Gly Leu Arg Ala Glu Gly
His Pro Asp Ser Leu Lys Asp Asn Ser 385 390 395 400 Ser Cys Ser Val
Met Ser Glu Glu Pro Glu Gly Arg Ser Tyr Ser Thr 405 410 415 Leu Thr
Thr Val Arg Glu Ile Glu Thr Gln Thr Glu Leu Leu Ser Pro 420 425 430
Gly Ser Gly Arg Ala Glu Glu Glu Glu Asp Gln Asp Glu Gly Ile Lys 435
440 445 Gln Ala Met Asn His Phe Val Gln Glu Asn Gly Thr Leu Arg Ala
Lys 450 455 460 Pro Thr Gly Asn Gly Ile Tyr Ile Asn Gly Arg Gly His
Leu Val 465 470 475 <210> SEQ ID NO 36 <211> LENGTH:
314 <212> TYPE: PRT <213> ORGANISM: Homo sapiens
<400> SEQUENCE: 36 Gly Glu Leu Glu Thr Ser Asp Val Val Thr
Val Val Leu Gly Gln Asp 1 5 10 15 Ala Lys Leu Pro Cys Phe Tyr Arg
Gly Asp Ser Gly Glu Gln Val Gly
20 25 30 Gln Val Ala Trp Ala Arg Val Asp Ala Gly Glu Gly Ala Gln
Glu Leu 35 40 45 Ala Leu Leu His Ser Lys Tyr Gly Leu His Val Ser
Pro Ala Tyr Glu 50 55 60 Gly Arg Val Glu Gln Pro Pro Pro Pro Arg
Asn Pro Leu Asp Gly Ser 65 70 75 80 Val Leu Leu Arg Asn Ala Val Gln
Ala Asp Glu Gly Glu Tyr Glu Cys 85 90 95 Arg Val Ser Thr Phe Pro
Ala Gly Ser Phe Gln Ala Arg Leu Arg Leu 100 105 110 Arg Val Leu Val
Pro Pro Leu Pro Ser Leu Asn Pro Gly Pro Ala Leu 115 120 125 Glu Glu
Gly Gln Gly Leu Thr Leu Ala Ala Ser Cys Thr Ala Glu Gly 130 135 140
Ser Pro Ala Pro Ser Val Thr Trp Asp Thr Glu Val Lys Gly Thr Thr 145
150 155 160 Ser Ser Arg Ser Phe Lys His Ser Arg Ser Ala Ala Val Thr
Ser Glu 165 170 175 Phe His Leu Val Pro Ser Arg Ser Met Asn Gly Gln
Pro Leu Thr Cys 180 185 190 Val Val Ser His Pro Gly Leu Leu Gln Asp
Gln Arg Ile Thr His Ile 195 200 205 Leu His Val Ser Phe Leu Ala Glu
Ala Ser Val Arg Gly Leu Glu Asp 210 215 220 Gln Asn Leu Trp His Ile
Gly Arg Glu Gly Ala Met Leu Lys Cys Leu 225 230 235 240 Ser Glu Gly
Gln Pro Pro Pro Ser Tyr Asn Trp Thr Arg Leu Asp Gly 245 250 255 Pro
Leu Pro Ser Gly Val Arg Val Asp Gly Asp Thr Leu Gly Phe Pro 260 265
270 Pro Leu Thr Thr Glu His Ser Gly Ile Tyr Val Cys His Val Ser Asn
275 280 285 Glu Phe Ser Ser Arg Asp Ser Gln Val Thr Val Asp Val Leu
Asp Pro 290 295 300 Gln Glu Asp Ser Gly Lys Gln Val Asp Leu 305 310
<210> SEQ ID NO 37 <211> LENGTH: 25 <212> TYPE:
PRT <213> ORGANISM: Homo sapiens <400> SEQUENCE: 37 Val
Ser Ala Ser Val Val Val Val Gly Val Ile Ala Ala Leu Leu Phe 1 5 10
15 Cys Leu Leu Val Val Val Val Val Leu 20 25 <210> SEQ ID NO
38 <211> LENGTH: 140 <212> TYPE: PRT <213>
ORGANISM: Homo sapiens <400> SEQUENCE: 38 Met Ser Arg Tyr His
Arg Arg Lys Ala Gln Gln Met Thr Gln Lys Tyr 1 5 10 15 Glu Glu Glu
Leu Thr Leu Thr Arg Glu Asn Ser Ile Arg Arg Leu His 20 25 30 Ser
His His Thr Asp Pro Arg Ser Gln Pro Glu Glu Ser Val Gly Leu 35 40
45 Arg Ala Glu Gly His Pro Asp Ser Leu Lys Asp Asn Ser Ser Cys Ser
50 55 60 Val Met Ser Glu Glu Pro Glu Gly Arg Ser Tyr Ser Thr Leu
Thr Thr 65 70 75 80 Val Arg Glu Ile Glu Thr Gln Thr Glu Leu Leu Ser
Pro Gly Ser Gly 85 90 95 Arg Ala Glu Glu Glu Glu Asp Gln Asp Glu
Gly Ile Lys Gln Ala Met 100 105 110 Asn His Phe Val Gln Glu Asn Gly
Thr Leu Arg Ala Lys Pro Thr Gly 115 120 125 Asn Gly Ile Tyr Ile Asn
Gly Arg Gly His Leu Val 130 135 140 <210> SEQ ID NO 39
<400> SEQUENCE: 39 000 3 <210> SEQ ID NO 40 <400>
SEQUENCE: 40 000 3 <210> SEQ ID NO 41 <211> LENGTH:
2510 <212> TYPE: DNA <213> ORGANISM: Homo sapiens
<400> SEQUENCE: 41 caaaggcaca acgtccagcc gttccttcaa
gcactcccgc tctgctgccg tcacctcaga 60 gttccacttg gtgcctagcc
gcagcatgaa tgggcagcca ctgacttgtg tggtgtccca 120 tcctggcctg
ctccaggacc aaaggatcac ccacatcctc cacgtgtcct tccttgctga 180
ggcctctgtg aggggccttg aagaccaaaa tctgtggcac attggcagag aaggagctat
240 gctcaagtgc ctgagtgaag ggcagccccc tccctcatac aactggacac
ggctggatgg 300 gcctctgccc agtggggtac gagtggatgg ggacactttg
ggctttcccc cactgaccac 360 tgagcacagc ggcatctacg tctgccatgt
cagcaatgag ttctcctcaa gggattctca 420 ggtcactgtg gatgttcttg
cagaccccca ggaagactct gggaagcagg tggacctagt 480 gtcagcctcg
gtggtggtgg tgggtgtgat cgccgcactc ttgttctgcc ttctggtggt 540
ggtggtggtg ctcatgtccc gataccatcg gcgcaaggcc cagcagatga cccagaaata
600 tgaggaggag ctgaccctga ccagggagaa ctccatccgg aggctgcatt
cccatcacac 660 ggaccccagg agccagagtg aagagcccga gggccgcagt
tactccacgc tgaccacggt 720 gagggagata gaaacacaga ctgaactgct
gtctccaggc tctgggcggg ccgaggagga 780 ggaagatcag gatgaaggca
tcaaacaggc catgaaccat tttgttcagg agaatgggac 840 cctacgggcc
aagcccacgg gcaatggcat ctacatcaat gggcggggac acctggtctg 900
acccaggcct gcctcccttc cctaggcctg gctccttctg ttgacatggg agattttagc
960 tcatcttggg ggcctcctta aacaccccca tttcttgcgg aagatgctcc
ccatcccact 1020 gactgcttga cctttacctc caacccttct gttcatcggg
agggctccac caattgagtc 1080 tctcccacca tgcatgcagg tcactgtgtg
tgtgcatgtg tgcctgtgtg agtgttgact 1140 gactgtgtgt gtgtggaggg
gtgactgtcc gtggaggggt gactgtgtcc gtggtgtgta 1200 ttatgctgtc
atatcagagt caagtgaact gtggtgtatg tgccacggga tttgagtggt 1260
tgcgtgggca acactgtcag ggtttggcgt gtgtgtcatg tggctgtgtg tgacctctgc
1320 ctgaaaaagc aggtattttc tcagacccca gagcagtatt aatgatgcag
aggttggagg 1380 agagaggtgg agactgtggc tcagacccag gtgtgcgggc
atagctggag ctggaatctg 1440 cctccggtgt gagggaacct gtctcctacc
acttcggagc catgggggca agtgtgaagc 1500 agccagtccc tgggtcagcc
agaggcttga actgttacag aagccctctg ccctctggtg 1560 gcctctgggc
ctgctgcatg tacatatttt ctgtaaatat acatgcgccg ggagcttctt 1620
gcaggaatac tgctccgaat cacttttaat ttttttcttt tttttttctt gccctttcca
1680 ttagttgtat tttttattta tttttatttt tatttttttt tagagatgga
gtctcactat 1740 gttgctcagg ctggccttga actcctgggc tcaagcaatc
ctcctgcctc agcctcccta 1800 gtagctggga ctttaagtgt acaccactgt
gcctgctttg aatcctttac gaagagaaaa 1860 aaaaaattaa agaaagcctt
tagatttatc caatgtttac tactgggatt gcttaaagtg 1920 aggcccctcc
aacaccaggg ggttaattcc tgtgattgtg aaaggggcta cttccaaggc 1980
atcttcatgc aggcagcccc ttgggagggc acctgagagc tggtagagtc tgaaattagg
2040 gatgtgagcc tggtgacaag ggctcctgtt caatagtggt gttggggaga
gagagagcag 2100 tgattataga ccgagagagt aggagttgag gtgaggtgaa
ggaggtgctg ggggtgagaa 2160 tgtcgccttt ccccctgggt tttggatcac
taattcaagg ctcttctgga tgtttctctg 2220 ggttggggct ggagttcaat
gaggtttatt tttagctggc ccacccagat acactcagcc 2280 agaataccta
gatttagtac ccaaactctt cttagtctga aatctgctgg atttctggcc 2340
taagggagag gctcccatcc ttcgttcccc agccagccta ggacttcgaa tgtggagcct
2400 gaagatctaa gatcctaaca tgtacatttt atgtaaatat gtgcatattt
gtacataaaa 2460 tgatattctg tttttaaata aacagacaaa acttgaaaaa
aaaaaaaaaa 2510 <210> SEQ ID NO 42 <211> LENGTH: 897
<212> TYPE: DNA <213> ORGANISM: Homo sapiens
<400> SEQUENCE: 42 aaaggcacaa cgtccagccg ttccttcaag
cactcccgct ctgctgccgt cacctcagag 60 ttccacttgg tgcctagccg
cagcatgaat gggcagccac tgacttgtgt ggtgtcccat 120 cctggcctgc
tccaggacca aaggatcacc cacatcctcc acgtgtcctt ccttgctgag 180
gcctctgtga ggggccttga agaccaaaat ctgtggcaca ttggcagaga aggagctatg
240 ctcaagtgcc tgagtgaagg gcagccccct ccctcataca actggacacg
gctggatggg 300 cctctgccca gtggggtacg agtggatggg gacactttgg
gctttccccc actgaccact 360 gagcacagcg gcatctacgt ctgccatgtc
agcaatgagt tctcctcaag ggattctcag 420 gtcactgtgg atgttcttgc
agacccccag gaagactctg ggaagcaggt ggacctagtg 480 tcagcctcgg
tggtggtggt gggtgtgatc gccgcactct tgttctgcct tctggtggtg 540
gtggtggtgc tcatgtcccg ataccatcgg cgcaaggccc agcagatgac ccagaaatat
600 gaggaggagc tgaccctgac cagggagaac tccatccgga ggctgcattc
ccatcacacg 660 gaccccagga gccagagtga agagcccgag ggccgcagtt
actccacgct gaccacggtg 720 agggagatag aaacacagac tgaactgctg
tctccaggct ctgggcgggc cgaggaggag 780 gaagatcagg atgaaggcat
caaacaggcc atgaaccatt ttgttcagga gaatgggacc 840 ctacgggcca
agcccacggg caatggcatc tacatcaatg ggcggggaca cctggtc 897 <210>
SEQ ID NO 43
<211> LENGTH: 299 <212> TYPE: PRT <213> ORGANISM:
Homo sapiens <400> SEQUENCE: 43 Lys Gly Thr Thr Ser Ser Arg
Ser Phe Lys His Ser Arg Ser Ala Ala 1 5 10 15 Val Thr Ser Glu Phe
His Leu Val Pro Ser Arg Ser Met Asn Gly Gln 20 25 30 Pro Leu Thr
Cys Val Val Ser His Pro Gly Leu Leu Gln Asp Gln Arg 35 40 45 Ile
Thr His Ile Leu His Val Ser Phe Leu Ala Glu Ala Ser Val Arg 50 55
60 Gly Leu Glu Asp Gln Asn Leu Trp His Ile Gly Arg Glu Gly Ala Met
65 70 75 80 Leu Lys Cys Leu Ser Glu Gly Gln Pro Pro Pro Ser Tyr Asn
Trp Thr 85 90 95 Arg Leu Asp Gly Pro Leu Pro Ser Gly Val Arg Val
Asp Gly Asp Thr 100 105 110 Leu Gly Phe Pro Pro Leu Thr Thr Glu His
Ser Gly Ile Tyr Val Cys 115 120 125 His Val Ser Asn Glu Phe Ser Ser
Arg Asp Ser Gln Val Thr Val Asp 130 135 140 Val Leu Ala Asp Pro Gln
Glu Asp Ser Gly Lys Gln Val Asp Leu Val 145 150 155 160 Ser Ala Ser
Val Val Val Val Gly Val Ile Ala Ala Leu Leu Phe Cys 165 170 175 Leu
Leu Val Val Val Val Val Leu Met Ser Arg Tyr His Arg Arg Lys 180 185
190 Ala Gln Gln Met Thr Gln Lys Tyr Glu Glu Glu Leu Thr Leu Thr Arg
195 200 205 Glu Asn Ser Ile Arg Arg Leu His Ser His His Thr Asp Pro
Arg Ser 210 215 220 Gln Ser Glu Glu Pro Glu Gly Arg Ser Tyr Ser Thr
Leu Thr Thr Val 225 230 235 240 Arg Glu Ile Glu Thr Gln Thr Glu Leu
Leu Ser Pro Gly Ser Gly Arg 245 250 255 Ala Glu Glu Glu Glu Asp Gln
Asp Glu Gly Ile Lys Gln Ala Met Asn 260 265 270 His Phe Val Gln Glu
Asn Gly Thr Leu Arg Ala Lys Pro Thr Gly Asn 275 280 285 Gly Ile Tyr
Ile Asn Gly Arg Gly His Leu Val 290 295 <210> SEQ ID NO 44
<400> SEQUENCE: 44 000 3 <210> SEQ ID NO 45 <400>
SEQUENCE: 45 000 3 <210> SEQ ID NO 46 <400> SEQUENCE:
46 000 3 <210> SEQ ID NO 47 <400> SEQUENCE: 47 000 3
<210> SEQ ID NO 48 <400> SEQUENCE: 48 000 3 <210>
SEQ ID NO 49 <400> SEQUENCE: 49 000 3 <210> SEQ ID NO
50 <400> SEQUENCE: 50 000 3 <210> SEQ ID NO 51
<211> LENGTH: 3114 <212> TYPE: DNA <213>
ORGANISM: Homo sapiens <400> SEQUENCE: 51 cttaatgttg
gaagtctctt agtcctatga gagtgtgtag cagtttgtcc ctgagctcta 60
gcttctttaa atgaagctga gtctctgggc aacatcttta gggagagagg tacaaaaggt
120 tcctggacct tctcaacaca gggagcctgc ataatgatgc aagagcagca
acctcaaagt 180 acagagaaaa gaggctggtt gtccctgaga ctctggtctg
tggctgggat ttccattgca 240 ctcctcagtg cttgcttcat tgtgagctgt
gtagtaactt accattttac atatggtgaa 300 actggcaaaa ggctgtctga
actacactca tatcattcaa gtctcacctg cttcagtgaa 360 gggacaaagg
tgccagcctg gggatgttgc ccagcttctt ggaagtcatt tggttccagt 420
tgctacttca tttccagtga agagaaggtt tggtctaaga gtgagcagaa ctgtgttgag
480 atgggagcac atttggttgt gttcaacaca gaagcagagc agaatttcat
tgtccagcag 540 ctgaatgagt cattttctta ttttctgggg ctttcagacc
cacaaggtaa taataattgg 600 caatggattg ataagacacc ttatgagaaa
aatgtcagat tttggcacct aggtgagccc 660 aatcattctg cagagcaatg
tgcttcaata gtcttctgga aacctacagg atggggctgg 720 aatgatgtta
tctgtgaaac tagaaggaat tcaatatgtg agatgaataa gatttaccta 780
tgagtagaag cttaattgga aagaagagaa gaattactga cgtaattttt tccctgacgt
840 ctttaaaatt gaaccctatc atgaaatgat aatttcttcc tgaatttaca
cataatcctt 900 atgttataga ggttcacaga aatggaaaga tacctgtttc
cctttaatca atcttctcgt 960 ttcctctttt ccattaatga tagaatgcac
ccttcctctc tttgttccat tctttcactt 1020 gttattcatt tttttctttc
ttcacacttc attacacaaa tatttattgt ttcagagact 1080 gtactatttt
gtttgttaga agatttataa ggcagtatct tttgaaaatt atgactttcc 1140
ttcctcaata taccataaag aaatcttttt ggtcaagatg gtagttggaa ctacaatcat
1200 ctgaaggcct gacaagagtt gaaagacatg ttttctagat ggctcactca
catggctggc 1260 aacttggtgt tggctattaa tgtaacctgg aaataaattt
tattctgcag ttagggattt 1320 ggcattttat atatgttgat tcaatcaagt
ttggcaagca gggtgttcga tactgctata 1380 tcctgtattc ttggtttatt
tgttttattt ctgagaaata tgtgttaaga tctctcgctg 1440 attgggaatt
tgtctatttc tcatttaaat tttgtcaaat ctttctttgc ttgcaagcat 1500
ttcttgttac ccaaatctaa cctattcctg aaaatatgat ggttagcaaa gtttgagata
1560 actagagcct gtaatccatc attttaaatg gcaatgataa tgacagttta
tttttatgtt 1620 atataaaaac ctcaacaaat tttccaaaca attaccaaaa
tggtcattaa tctgtatcca 1680 caaaggattt ctgcattaca tactttaaaa
caaattacct aattatttag tgcatattaa 1740 acttattggt gggcatgact
atatgcaaca gttgcatgat atatgataca aattatgtta 1800 ttcttttcca
ttgcactgaa aataccataa tataaagaag aatcccatca tccaaattga 1860
gcctatattg attgatactc agaagaatct ggcagtagga gcctataaag ggataagcaa
1920 ttgggaaagg attgggaagt tggtagtact gaacatcttc tcacctggac
tcatgagcaa 1980 cttgaatagt tgtaactgtg atgcatatgt agattctaac
acatttttcc cccttgaata 2040 gaaatttggc acaacaattt tttaaattaa
tttagcaaat atttggatat taaagcttct 2100 tatagaaaga gatacctgta
tatttaagcc atgatgaggt atatacaatg ttataattat 2160 tacttgtaca
tggcaaatta atttttttat cattgtggag tcactttctt taaatttagt 2220
aatgcctttg gctttaattt ttctcctgat attaaaatag atacagtaac tttcattatg
2280 ttagtgctgt aaaatttttt tttccatctt ctatttttga ccatttttat
tccacatgtg 2340 ctcttaataa gtagcatata gttaaatttt aaaaaatcca
atatggcaat caccttttag 2400 gttaaaaatt taatccattt acatttgtga
caattcgaca tatatatggt tctaaatcta 2460 tcatcttact aggtggtttc
catttcctct gctccaaaat atttttttta cagcttataa 2520 cacaactttt
attagaaaag ttatacataa cacagcatca actattttca agaacccaat 2580
aagcaacaaa aaccagacta acaaaatgtg taacaagaaa ctaatgacct ttctaaaatc
2640 aaacattcaa ttatctacaa tgtctattta caaacaggga aaactccatg
gtttacaggc 2700 atgtcatatt gaaaataaag ctgcaatagc tttttataca
attatcgctc tcaagaaaat 2760 gaatcattaa gacagtaatt aggagttcac
aaatttaaaa catttcacgt aattttaaat 2820 tattgtcttc aataatttta
aattattgaa gtctgagttt caaaagtgat tttttcccac 2880 aaaggtgcca
acacttaagc tagagctttc agtgttaact ttgccctaaa agttaagaca 2940
tattctgaga atcataatag tcacatgatt tctgatgcta tctgctctgt taataacaaa
3000 gatttcacac atgaatacct atgtaacaaa tctccatgtt ctacacatat
accccagaac 3060 ttaaagtata ataataataa aacatagcaa agcctttaaa
aaaaaaaaaa aaaa 3114 <210> SEQ ID NO 52 <211> LENGTH:
627 <212> TYPE: DNA <213> ORGANISM: Homo sapiens
<400> SEQUENCE: 52 atgatgcaag agcagcaacc tcaaagtaca
gagaaaagag gctggttgtc cctgagactc 60 tggtctgtgg ctgggatttc
cattgcactc ctcagtgctt gcttcattgt gagctgtgta 120 gtaacttacc
attttacata tggtgaaact ggcaaaaggc tgtctgaact acactcatat 180
cattcaagtc tcacctgctt cagtgaaggg acaaaggtgc cagcctgggg atgttgccca
240 gcttcttgga agtcatttgg ttccagttgc tacttcattt ccagtgaaga
gaaggtttgg 300 tctaagagtg agcagaactg tgttgagatg ggagcacatt
tggttgtgtt caacacagaa 360 gcagagcaga atttcattgt ccagcagctg
aatgagtcat tttcttattt tctggggctt 420 tcagacccac aaggtaataa
taattggcaa tggattgata agacacctta tgagaaaaat 480 gtcagatttt
ggcacctagg tgagcccaat cattctgcag agcaatgtgc ttcaatagtc 540
ttctggaaac ctacaggatg gggctggaat gatgttatct gtgaaactag aaggaattca
600
atatgtgaga tgaataagat ttaccta 627 <210> SEQ ID NO 53
<211> LENGTH: 209 <212> TYPE: PRT <213> ORGANISM:
Homo sapiens <400> SEQUENCE: 53 Met Met Gln Glu Gln Gln Pro
Gln Ser Thr Glu Lys Arg Gly Trp Leu 1 5 10 15 Ser Leu Arg Leu Trp
Ser Val Ala Gly Ile Ser Ile Ala Leu Leu Ser 20 25 30 Ala Cys Phe
Ile Val Ser Cys Val Val Thr Tyr His Phe Thr Tyr Gly 35 40 45 Glu
Thr Gly Lys Arg Leu Ser Glu Leu His Ser Tyr His Ser Ser Leu 50 55
60 Thr Cys Phe Ser Glu Gly Thr Lys Val Pro Ala Trp Gly Cys Cys Pro
65 70 75 80 Ala Ser Trp Lys Ser Phe Gly Ser Ser Cys Tyr Phe Ile Ser
Ser Glu 85 90 95 Glu Lys Val Trp Ser Lys Ser Glu Gln Asn Cys Val
Glu Met Gly Ala 100 105 110 His Leu Val Val Phe Asn Thr Glu Ala Glu
Gln Asn Phe Ile Val Gln 115 120 125 Gln Leu Asn Glu Ser Phe Ser Tyr
Phe Leu Gly Leu Ser Asp Pro Gln 130 135 140 Gly Asn Asn Asn Trp Gln
Trp Ile Asp Lys Thr Pro Tyr Glu Lys Asn 145 150 155 160 Val Arg Phe
Trp His Leu Gly Glu Pro Asn His Ser Ala Glu Gln Cys 165 170 175 Ala
Ser Ile Val Phe Trp Lys Pro Thr Gly Trp Gly Trp Asn Asp Val 180 185
190 Ile Cys Glu Thr Arg Arg Asn Ser Ile Cys Glu Met Asn Lys Ile Tyr
195 200 205 Leu <210> SEQ ID NO 54 <211> LENGTH: 48
<212> TYPE: PRT <213> ORGANISM: Homo sapiens
<400> SEQUENCE: 54 Met Met Gln Glu Gln Gln Pro Gln Ser Thr
Glu Lys Arg Gly Trp Leu 1 5 10 15 Ser Leu Arg Leu Trp Ser Val Ala
Gly Ile Ser Ile Ala Leu Leu Ser 20 25 30 Ala Cys Phe Ile Val Ser
Cys Val Val Thr Tyr His Phe Thr Tyr Gly 35 40 45 <210> SEQ ID
NO 55 <211> LENGTH: 161 <212> TYPE: PRT <213>
ORGANISM: Homo sapiens <400> SEQUENCE: 55 Glu Thr Gly Lys Arg
Leu Ser Glu Leu His Ser Tyr His Ser Ser Leu 1 5 10 15 Thr Cys Phe
Ser Glu Gly Thr Lys Val Pro Ala Trp Gly Cys Cys Pro 20 25 30 Ala
Ser Trp Lys Ser Phe Gly Ser Ser Cys Tyr Phe Ile Ser Ser Glu 35 40
45 Glu Lys Val Trp Ser Lys Ser Glu Gln Asn Cys Val Glu Met Gly Ala
50 55 60 His Leu Val Val Phe Asn Thr Glu Ala Glu Gln Asn Phe Ile
Val Gln 65 70 75 80 Gln Leu Asn Glu Ser Phe Ser Tyr Phe Leu Gly Leu
Ser Asp Pro Gln 85 90 95 Gly Asn Asn Asn Trp Gln Trp Ile Asp Lys
Thr Pro Tyr Glu Lys Asn 100 105 110 Val Arg Phe Trp His Leu Gly Glu
Pro Asn His Ser Ala Glu Gln Cys 115 120 125 Ala Ser Ile Val Phe Trp
Lys Pro Thr Gly Trp Gly Trp Asn Asp Val 130 135 140 Ile Cys Glu Thr
Arg Arg Asn Ser Ile Cys Glu Met Asn Lys Ile Tyr 145 150 155 160 Leu
<210> SEQ ID NO 56 <400> SEQUENCE: 56 000 3 <210>
SEQ ID NO 57 <400> SEQUENCE: 57 000 3 <210> SEQ ID NO
58 <400> SEQUENCE: 58 000 3 <210> SEQ ID NO 59
<400> SEQUENCE: 59 000 3 <210> SEQ ID NO 60 <211>
LENGTH: 209 <212> TYPE: PRT <213> ORGANISM: Mus sp.
<400> SEQUENCE: 60 Met Val Gln Glu Arg Gln Ser Gln Gly Lys
Gly Val Cys Trp Thr Leu 1 5 10 15 Arg Leu Trp Ser Ala Ala Val Ile
Ser Met Leu Leu Leu Ser Thr Cys 20 25 30 Phe Ile Ala Ser Cys Val
Val Thr Tyr Gln Phe Ile Met Asp Gln Pro 35 40 45 Ser Arg Arg Leu
Tyr Glu Leu His Thr Tyr His Ser Ser Leu Thr Cys 50 55 60 Phe Ser
Glu Gly Thr Met Val Ser Glu Lys Met Trp Gly Cys Cys Pro 65 70 75 80
Asn His Trp Lys Ser Phe Gly Ser Ser Cys Tyr Leu Ile Ser Thr Lys 85
90 95 Glu Asn Phe Trp Ser Thr Ser Glu Gln Asn Cys Val Gln Met Gly
Ala 100 105 110 His Leu Val Val Ile Asn Thr Glu Ala Glu Gln Asn Phe
Ile Thr Gln 115 120 125 Gln Leu Asn Glu Ser Leu Ser Tyr Phe Leu Gly
Leu Ser Asp Pro Gln 130 135 140 Gly Asn Gly Lys Trp Gln Trp Ile Asp
Asp Thr Pro Phe Ser Gln Asn 145 150 155 160 Val Arg Phe Trp His Pro
His Glu Pro Asn Leu Pro Glu Glu Arg Cys 165 170 175 Val Ser Ile Val
Tyr Trp Asn Pro Ser Lys Trp Gly Trp Asn Asp Val 180 185 190 Phe Cys
Asp Ser Lys His Asn Ser Ile Cys Glu Met Lys Lys Ile Tyr 195 200 205
Leu <210> SEQ ID NO 61 <211> LENGTH: 821 <212>
TYPE: DNA <213> ORGANISM: Mus sp. <220> FEATURE:
<221> NAME/KEY: misc_feature <222> LOCATION:
(788)..(788) <223> OTHER INFORMATION: unsure <400>
SEQUENCE: 61 gaactccccg gtgtcgaccc cgcgtcccga ttggcccgct ctgtggcatt
taactcaagt 60 gtgtgtggaa gttgattctg aactctggcc tctttgacag
aagccaggtc cctgagtcgt 120 attttggaga cagatgcaag aaacccctga
ccttctgaac atacacctca acaatggtgc 180 aggaaagaca atcccaaggg
aagggagtct gctggaccct gagactctgg tcagctgctg 240 tgatttccat
gttactcttg agtacctgtt tcattgcgag ctgtgtggtg acttaccaat 300
ttattatgga ccagcccagt agaagactat atgaacttca cacataccat tccagtctca
360 cctgcttcag tgaagggact atggtgtcag aaaaaatgtg gggatgctgc
ccaaatcact 420 ggaagtcatt tggctccagc tgctacctca tttctaccaa
ggagaacttc tggagcacca 480 gtgagcagaa ctgtgttcag atgggggctc
atctggtggt gatcaatact gaagcggagc 540 agaatttcat cacccagcag
ctgaatgagt cactttctta cttcctgggt ctttcggatc 600 ccaaggtaat
ggcaaatggc aatggatcga tgatactcct ttcagtcaaa atgtcaggtt 660
ctggcacccc catgaaccca atcttccaga agagcggtgt gtttcaatag tttactggaa
720 tccttcgaaa tggggctggg aatgatgttt tctgtgatag taaacacaat
tcaatatgtg 780 aaatgaanaa gattacctat gaatgcctgt tattcttaat a 821
<210> SEQ ID NO 62 <211> LENGTH: 534 <212> TYPE:
DNA <213> ORGANISM: Mus sp. <400> SEQUENCE: 62
atggtgcagg aaagacaatc ccaagggaag ggagtctgct ggaccctgag actctggtca
60 gctgctgtga tttccatgtt actcttgagt acctgtttca ttgcgagctg
tgtggtgact 120 taccaattta ttatggacca gcccagtaga agactatatg
aacttcacac ataccattcc 180 agtctcacct gcttcagtga agggactatg
gtgtcagaaa aaatgtgggg atgctgccca 240 aatcactgga agtcatttgg
ctccagctgc tacctcattt ctaccaagga gaacttctgg 300 agcaccagtg
agcagaactg tgttcagatg ggggctcatc tggtggtgat caatactgaa 360
gcggagcaga atttcatcac ccagcagctg aatgagtcac tttcttactt cctgggtctt
420 tcggatccca aggtaatggc aaatggcaat ggatcgatga tactcctttc
agtcaaaatg 480 tcaggttctg gcacccccat gaacccaatc ttccagaaga
gcggtgtgtt tcaa 534
<210> SEQ ID NO 63 <211> LENGTH: 178 <212> TYPE:
PRT <213> ORGANISM: Mus sp. <400> SEQUENCE: 63 Met Val
Gln Glu Arg Gln Ser Gln Gly Lys Gly Val Cys Trp Thr Leu 1 5 10 15
Arg Leu Trp Ser Ala Ala Val Ile Ser Met Leu Leu Leu Ser Thr Cys 20
25 30 Phe Ile Ala Ser Cys Val Val Thr Tyr Gln Phe Ile Met Asp Gln
Pro 35 40 45 Ser Arg Arg Leu Tyr Glu Leu His Thr Tyr His Ser Ser
Leu Thr Cys 50 55 60 Phe Ser Glu Gly Thr Met Val Ser Glu Lys Met
Trp Gly Cys Cys Pro 65 70 75 80 Asn His Trp Lys Ser Phe Gly Ser Ser
Cys Tyr Leu Ile Ser Thr Lys 85 90 95 Glu Asn Phe Trp Ser Thr Ser
Glu Gln Asn Cys Val Gln Met Gly Ala 100 105 110 His Leu Val Val Ile
Asn Thr Glu Ala Glu Gln Asn Phe Ile Thr Gln 115 120 125 Gln Leu Asn
Glu Ser Leu Ser Tyr Phe Leu Gly Leu Ser Asp Pro Lys 130 135 140 Val
Met Ala Asn Gly Asn Gly Ser Met Ile Leu Leu Ser Val Lys Met 145 150
155 160 Ser Gly Ser Gly Thr Pro Met Asn Pro Ile Phe Gln Lys Ser Gly
Val 165 170 175 Phe Gln <210> SEQ ID NO 64 <211>
LENGTH: 48 <212> TYPE: PRT <213> ORGANISM: Mus sp.
<400> SEQUENCE: 64 Met Val Gln Glu Arg Gln Ser Gln Gly Lys
Gly Val Cys Trp Thr Leu 1 5 10 15 Arg Leu Trp Ser Ala Ala Val Ile
Ser Met Leu Leu Leu Ser Thr Cys 20 25 30 Phe Ile Ala Ser Cys Val
Val Thr Tyr Gln Phe Ile Met Asp Gln Pro 35 40 45 <210> SEQ ID
NO 65 <211> LENGTH: 130 <212> TYPE: PRT <213>
ORGANISM: Mus sp. <400> SEQUENCE: 65 Ser Arg Arg Leu Tyr Glu
Leu His Thr Tyr His Ser Ser Leu Thr Cys 1 5 10 15 Phe Ser Glu Gly
Thr Met Val Ser Glu Lys Met Trp Gly Cys Cys Pro 20 25 30 Asn His
Trp Lys Ser Phe Gly Ser Ser Cys Tyr Leu Ile Ser Thr Lys 35 40 45
Glu Asn Phe Trp Ser Thr Ser Glu Gln Asn Cys Val Gln Met Gly Ala 50
55 60 His Leu Val Val Ile Asn Thr Glu Ala Glu Gln Asn Phe Ile Thr
Gln 65 70 75 80 Gln Leu Asn Glu Ser Leu Ser Tyr Phe Leu Gly Leu Ser
Asp Pro Lys 85 90 95 Val Met Ala Asn Gly Asn Gly Ser Met Ile Leu
Leu Ser Val Lys Met 100 105 110 Ser Gly Ser Gly Thr Pro Met Asn Pro
Ile Phe Gln Lys Ser Gly Val 115 120 125 Phe Gln 130 <210> SEQ
ID NO 66 <400> SEQUENCE: 66 000 3 <210> SEQ ID NO 67
<400> SEQUENCE: 67 000 3 <210> SEQ ID NO 68 <400>
SEQUENCE: 68 000 3 <210> SEQ ID NO 69 <400> SEQUENCE:
69 000 3 <210> SEQ ID NO 70 <400> SEQUENCE: 70 000 3
<210> SEQ ID NO 71 <211> LENGTH: 1252 <212> TYPE:
DNA <213> ORGANISM: Mus sp. <400> SEQUENCE: 71
cgaccccgcg tccgctgact tctgggtttg cagcattggc ccgctctgtg gcatttaact
60 caagtgtgtg tggaagttga ttctgaactc tggcctcttt gacagaagcc
aggtccctga 120 gtcgtatttt ggagacagat gcaagaaacc cctgaccttc
tgaacataca cctcaacaat 180 ggtgcaggaa agacaatccc aagggaaggg
agtctgctgg accctgagac tctggtcagc 240 tgctgtgatt tccatgttac
tcttgagtac ctgtttcatt gcgagctgtg tggtgactta 300 ccaatttatt
atggaccagc ccagtagaag actatatgaa cttcacacat accattccag 360
tctcacctgc ttcagtgaag ggactatggt gtcagaaaaa atgtggggat gctgcccaaa
420 tcactggaag tcatttggct ccagctgcta cctcatttct accaaggaga
acttctggag 480 caccagtgag cagaactgtg ttcagatggg ggctcatctg
gtggtgatca atactgaagc 540 ggagcagaat ttcatcaccc agcagctgaa
tgagtcactt tcttacttcc tgggtctttc 600 ggatccacaa ggtaatggca
aatggcaatg gatcgatgat actcctttca gtcaaaatgt 660 caggttctgg
cacccccatg aacccaatct tccagaagag cggtgtgttt caatagttta 720
ctggaatcct tcgaaatggg gctggaatga tgttttctgt gatagtaaac acaattcaat
780 atgtgaaatg aagaagattt acctatgagt gcctgttatt cattaatatc
tttaaagttc 840 agacctacca agaagccata acttcttggc ctgtacatct
gacagaggcc gttcttttcc 900 tagccactat tctttactca aacagaatga
gccctttctc cttctgatgg ttagagtttt 960 gtcaacttga cacaaactag
agtcacctgg ggagtaggat cttcagctaa ggaattgcct 1020 ctgtcagctt
gaccagtcag catgtctggg ggcattttct tgattaatga ttgttgtaag 1080
agggtccagg tggtaagcaa aggtgttaaa cccatgaaga gcaagccagg gagcatcatc
1140 catccatctc tgccctcagg tttctgcccc agggtcttgc cctggtttct
ttctatgaac 1200 tgctgttact tgaaagtata agatgaataa acaatttcat
ccaaaaaaaa aa 1252 <210> SEQ ID NO 72 <211> LENGTH: 627
<212> TYPE: DNA <213> ORGANISM: Mus sp. <400>
SEQUENCE: 72 atggtgcagg aaagacaatc ccaagggaag ggagtctgct ggaccctgag
actctggtca 60 gctgctgtga tttccatgtt actcttgagt acctgtttca
ttgcgagctg tgtggtgact 120 taccaattta ttatggacca gcccagtaga
agactatatg aacttcacac ataccattcc 180 agtctcacct gcttcagtga
agggactatg gtgtcagaaa aaatgtgggg atgctgccca 240 aatcactgga
agtcatttgg ctccagctgc tacctcattt ctaccaagga gaacttctgg 300
agcaccagtg agcagaactg tgttcagatg ggggctcatc tggtggtgat caatactgaa
360 gcggagcaga atttcatcac ccagcagctg aatgagtcac tttcttactt
cctgggtctt 420 tcggatccac aaggtaatgg caaatggcaa tggatcgatg
atactccttt cagtcaaaat 480 gtcaggttct ggcaccccca tgaacccaat
cttccagaag agcggtgtgt ttcaatagtt 540 tactggaatc cttcgaaatg
gggctggaat gatgttttct gtgatagtaa acacaattca 600 atatgtgaaa
tgaagaagat ttaccta 627 <210> SEQ ID NO 73 <211> LENGTH:
586 <212> TYPE: PRT <213> ORGANISM: Mus sp. <400>
SEQUENCE: 73 Met Glu Thr Val Ala Leu Gly Leu Asn Gly Leu Ala Arg
Gly Gly Leu 1 5 10 15 Asn Ser Glu Arg Gly Leu Asn Gly Leu Tyr Leu
Tyr Ser Gly Leu Tyr 20 25 30 Val Ala Leu Cys Tyr Ser Thr Arg Pro
Thr His Arg Leu Glu Ala Arg 35 40 45 Gly Leu Glu Thr Arg Pro Ser
Glu Arg Ala Leu Ala Ala Leu Ala Val 50 55 60 Ala Leu Ile Leu Glu
Ser Glu Arg Met Glu Thr Leu Glu Leu Glu Leu 65 70 75 80 Glu Ser Glu
Arg Thr His Arg Cys Tyr Ser Pro His Glu Ile Leu Glu 85 90 95 Ala
Leu Ala Ser Glu Arg Cys Tyr Ser Val Ala Leu Val Ala Leu Thr 100 105
110 His Arg Thr Tyr Arg Gly Leu Asn Pro His Glu Ile Leu Glu Met Glu
115 120 125 Thr Ala Ser Pro Gly Leu Asn Pro Arg Ser Glu Arg Ala Arg
Gly Ala 130 135 140 Arg Gly Leu Glu Thr Tyr Arg Gly Leu Leu Glu His
Ile Ser Thr His 145 150 155 160 Arg Thr Tyr Arg His Ile Ser Ser Glu
Arg Ser Glu Arg Leu Glu Thr 165 170 175
His Arg Cys Tyr Ser Pro His Glu Ser Glu Arg Gly Leu Gly Leu Tyr 180
185 190 Thr His Arg Met Glu Thr Val Ala Leu Ser Glu Arg Gly Leu Leu
Tyr 195 200 205 Ser Met Glu Thr Thr Arg Pro Gly Leu Tyr Cys Tyr Ser
Cys Tyr Ser 210 215 220 Pro Arg Ala Ser Asn His Ile Ser Thr Arg Pro
Leu Tyr Ser Ser Glu 225 230 235 240 Arg Pro His Glu Gly Leu Tyr Ser
Glu Arg Ser Glu Arg Cys Tyr Ser 245 250 255 Thr Tyr Arg Leu Glu Ile
Leu Glu Ser Glu Arg Thr His Arg Leu Tyr 260 265 270 Ser Gly Leu Ala
Ser Asn Pro His Glu Thr Arg Pro Ser Glu Arg Thr 275 280 285 His Arg
Ser Glu Arg Gly Leu Gly Leu Asn Ala Ser Asn Cys Tyr Ser 290 295 300
Val Ala Leu Gly Leu Asn Met Glu Thr Gly Leu Tyr Ala Leu Ala His 305
310 315 320 Ile Ser Leu Glu Val Ala Leu Val Ala Leu Ile Leu Glu Ala
Ser Asn 325 330 335 Thr His Arg Gly Leu Ala Leu Ala Gly Leu Gly Leu
Asn Ala Ser Asn 340 345 350 Pro His Glu Ile Leu Glu Thr His Arg Gly
Leu Asn Gly Leu Asn Leu 355 360 365 Glu Ala Ser Asn Gly Leu Ser Glu
Arg Leu Glu Ser Glu Arg Thr Tyr 370 375 380 Arg Pro His Glu Leu Glu
Gly Leu Tyr Leu Glu Ser Glu Arg Ala Ser 385 390 395 400 Pro Pro Arg
Gly Leu Asn Gly Leu Tyr Ala Ser Asn Gly Leu Tyr Leu 405 410 415 Tyr
Ser Thr Arg Pro Gly Leu Asn Thr Arg Pro Ile Leu Glu Ala Ser 420 425
430 Pro Ala Ser Pro Thr His Arg Pro Arg Pro His Glu Ser Glu Arg Gly
435 440 445 Leu Asn Ala Ser Asn Val Ala Leu Ala Arg Gly Pro His Glu
Thr Arg 450 455 460 Pro His Ile Ser Pro Arg His Ile Ser Gly Leu Pro
Arg Ala Ser Asn 465 470 475 480 Leu Glu Pro Arg Gly Leu Gly Leu Ala
Arg Gly Cys Tyr Ser Val Ala 485 490 495 Leu Ser Glu Arg Ile Leu Glu
Val Ala Leu Thr Tyr Arg Trp Ala Ser 500 505 510 Asn Pro Arg Ser Glu
Arg Leu Tyr Ser Thr Arg Pro Gly Leu Tyr Thr 515 520 525 Arg Pro Ala
Ser Asn Ala Ser Pro Val Ala Leu Phe Cys Tyr Ser Ala 530 535 540 Ser
Pro Ser Glu Arg Leu Tyr Ser His Ile Ser Ala Ser Asn Ser Glu 545 550
555 560 Arg Ile Leu Glu Cys Tyr Ser Gly Leu Met Glu Thr Leu Tyr Ser
Leu 565 570 575 Tyr Ser Ile Leu Glu Thr Tyr Arg Leu Glu 580 585
<210> SEQ ID NO 74 <400> SEQUENCE: 74 000 3 <210>
SEQ ID NO 75 <400> SEQUENCE: 75 000 3 <210> SEQ ID NO
76 <400> SEQUENCE: 76 000 3 <210> SEQ ID NO 77
<400> SEQUENCE: 77 000 3 <210> SEQ ID NO 78 <400>
SEQUENCE: 78 000 3 <210> SEQ ID NO 79 <400> SEQUENCE:
79 000 3 <210> SEQ ID NO 80 <400> SEQUENCE: 80 000 3
<210> SEQ ID NO 81 <211> LENGTH: 1202 <212> TYPE:
DNA <213> ORGANISM: Homo sapiens <400> SEQUENCE: 81
gtcgacccac gcgtccggaa accattccac aatcaccctc ctgaggaact cttagcactg
60 cataaagtgt tctgagtttg taatcagata ttgtcacact ggttccttca
aacagacatg 120 acaaggagct ggctttgggc taggctgctc cttgcctatg
attggggaag gttaaacccc 180 tacagggctt atgtatgtgg aaactgttgg
aacactgatt aaatgggatg gacttcactt 240 aacactcttg gatttccaat
attatgtttg agtaaaagaa ctgctatcca caaacaccat 300 taatccttta
gggaggcaga aaaggccaga atgcaaagcc atcttttcat tacactaggg 360
tctgtctttt tacttctctg ggcctttatc tggggagggc atgtttcccc cacttggaac
420 agtgagcctg gccaggacag taacctgtgg gcttgtgatg acattatttc
taatagggaa 480 tgggaaagga tgttagcttc tcaggtttta aagtgtcctg
gaggagaaga gaaaggacga 540 catgagaagg agacaatgaa gaagatgggt
gagggggaga tagtgtaaga ccctgagaat 600 ggcatagggt aaaactggga
cagagatact gtgggagaac gatagctgca gagggacaga 660 gggaggaagg
aaggagaaga gagggagata aaaacagttt ggagaaactc tcacaataca 720
ttcataagaa gacaaagaac ccaataaaaa tgggcaacag ataccacaga agatgatata
780 ttgagtggcc aataaataca taaaaatatg ctcaacatct ataattacca
gggaaatgca 840 aattaaaagc actgtgagat accactacac actgatgaga
atggctaaaa tcaaaaaaga 900 ccaaccagca ctttgggagg ccgaggtggg
cggatcatga ggtcaggagt ttgagactag 960 cctgaccaac atggtgaaac
cctgtctcta ctaaacatac aaaaattagc tgggggtggt 1020 ggcatgcgcc
tgtaattcca gctactcagg aggctgaggc aggagaatcg cttgaaccca 1080
ggaggcagag attacagtga gccgagatca tgcccttgca ctctagcctg ggtgacagag
1140 cgagactctg tcttaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa
aagggcggcc 1200 gc 1202 <210> SEQ ID NO 82 <211>
LENGTH: 255 <212> TYPE: DNA <213> ORGANISM: Homo
sapiens <400> SEQUENCE: 82 atgcaaagcc atcttttcat tacactaggg
tctgtctttt tacttctctg ggcctttatc 60 tggggagggc atgtttcccc
cacttggaac agtgagcctg gccaggacag taacctgtgg 120 gcttgtgatg
acattatttc taatagggaa tgggaaagga tgttagcttc tcaggtttta 180
aagtgtcctg gaggagaaga gaaaggacga catgagaagg agacaatgaa gaagatgggt
240 gagggggaga tagtg 255 <210> SEQ ID NO 83 <211>
LENGTH: 85 <212> TYPE: PRT <213> ORGANISM: Homo sapiens
<400> SEQUENCE: 83 Met Gln Ser His Leu Phe Ile Thr Leu Gly
Ser Val Phe Leu Leu Leu 1 5 10 15 Trp Ala Phe Ile Trp Gly Gly His
Val Ser Pro Thr Trp Asn Ser Glu 20 25 30 Pro Gly Gln Asp Ser Asn
Leu Trp Ala Cys Asp Asp Ile Ile Ser Asn 35 40 45 Arg Glu Trp Glu
Arg Met Leu Ala Ser Gln Val Leu Lys Cys Pro Gly 50 55 60 Gly Glu
Glu Lys Gly Arg His Glu Lys Glu Thr Met Lys Lys Met Gly 65 70 75 80
Glu Gly Glu Ile Val 85 <210> SEQ ID NO 84 <211> LENGTH:
23 <212> TYPE: PRT <213> ORGANISM: Homo sapiens
<400> SEQUENCE: 84 Met Gln Ser His Leu Phe Ile Thr Leu Gly
Ser Val Phe Leu Leu Leu 1 5 10 15 Trp Ala Phe Ile Trp Gly Gly 20
<210> SEQ ID NO 85 <211> LENGTH: 62 <212> TYPE:
PRT <213> ORGANISM: Homo sapiens <400> SEQUENCE: 85 His
Val Ser Pro Thr Trp Asn Ser Glu Pro Gly Gln Asp Ser Asn Leu 1 5 10
15 Trp Ala Cys Asp Asp Ile Ile Ser Asn Arg Glu Trp Glu Arg Met Leu
20 25 30 Ala Ser Gln Val Leu Lys Cys Pro Gly Gly Glu Glu Lys Gly
Arg His
35 40 45 Glu Lys Glu Thr Met Lys Lys Met Gly Glu Gly Glu Ile Val 50
55 60
* * * * *
References