U.S. patent application number 10/020269 was filed with the patent office on 2003-09-18 for identification of antigenic peptide sequences.
Invention is credited to Mitchell, William M., Stratton, Charles W..
Application Number | 20030175310 10/020269 |
Document ID | / |
Family ID | 46276209 |
Filed Date | 2003-09-18 |
United States Patent
Application |
20030175310 |
Kind Code |
A1 |
Mitchell, William M. ; et
al. |
September 18, 2003 |
Identification of antigenic peptide sequences
Abstract
Identification of linear amino acid antigenic sequences for the
production of both polyclonal and monoclonal antibodies to defined
antigenic domains is described. Also described are antigenic
peptides identified by the described methods and antibodies
thereto.
Inventors: |
Mitchell, William M.;
(Nashville, TN) ; Stratton, Charles W.;
(Nashville, TN) |
Correspondence
Address: |
CLARK & ELBING LLP
101 FEDERAL STREET
BOSTON
MA
02110
US
|
Family ID: |
46276209 |
Appl. No.: |
10/020269 |
Filed: |
December 14, 2001 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
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10020269 |
Dec 14, 2001 |
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09025596 |
Feb 18, 1998 |
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6340463 |
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09025596 |
Feb 18, 1998 |
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08911593 |
Aug 14, 1997 |
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60023921 |
Aug 14, 1996 |
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Current U.S.
Class: |
424/263.1 ;
530/350; 530/388.4; 536/23.7 |
Current CPC
Class: |
A61K 31/455 20130101;
A61K 31/455 20130101; A61K 2300/00 20130101; C07K 14/295 20130101;
A61K 45/06 20130101 |
Class at
Publication: |
424/263.1 ;
530/350; 530/388.4; 536/23.7 |
International
Class: |
A61K 039/118; C07H
021/04; C07K 014/295; C07K 016/12 |
Claims
1. A substantially pure immunogenic polypeptide having a sequence
consisting essentially of SEQ ID NO: 81 or a subsequence of SEQ ID
NO: 81 that comprises amino acids 2 to 13 of SEQ ID NO: 101.
2. The substantially pure immunogenic polypeptide of claim 1,
wherein said sequence consists essentially of SEQ ID NO: 81 or
amino acids 2 to 13 of SEQ ID NO: 101.
3. A substantially pure nucleic acid encoding an immunogenic
polypeptide having a sequence consisting essentially of SEQ ID NO:
81 or a subsequence of SEQ ID NO: 81 that comprises amino acids 2
to 13 of SEQ ID NO: 101.
4. The substantially pure nucleic acid of claim 6, wherein said
sequence consists essentially of SEQ ID NO: 81 or amino acids 2 to
13 of SEQ ID NO: 101.
5. A substantially pure antibody that specifically binds to a
polypeptide having a sequence consisting essentially of SEQ ID NO:
81 or a subsequence of SEQ ID NO: 81 that comprises amino acids 2
to 13 of SEQ ID NO: 101.
Description
CROSS REFERENCE TO RELATED APPLICATIONS
[0001] This application is a continuation of and claims priority
from U.S. patent application Ser. No. 09/025,596, filed Feb. 18,
1998 (now allowed), which is a continuation-in-part of U.S. utility
application U.S. Pat. Ser. No. 08/911,593, filed Aug. 14, 1997 (now
abandoned), which claims benefit from U.S. provisional application
U.S. Pat. Ser. No. 60/023,921, filed Aug. 14, 1996 (now abandoned),
the entire teachings of which are incorporated herein by
reference.
BACKGROUND OF THE INVENTION
[0002] Antibodies are widely used in diagnostic assays in both
human and veterinary medicine. Uses include enzyme-linked
immunosorbent analysis (ELISA), quantitative antigen capture
analysis, radioisotope-tagged reagents for in vivo localization of
target antigens, and for in vivo localization of cytotoxic agents
to target cells (i.e., immunotoxic therapy). The minimum epitope
size for protein antigens is generally considered to be 5-6 amino
acids, either as a linear sequence or as non-contiguous amino acids
whose spatial placement defines the epitope (i.e., conformational
epitope). Specificity is provided by the large number of potential
amino acid epitopic sequences possible for a minimum epitope (i.e.,
5.sup.20).
[0003] Most commonly, large antigens or microbial organisms are
used to induce antibody responses in order to insure the
presentation of good antigenic sequences in the host animal. The
use of these multivalent antigens for the production of polyclonal
antibodies generally requires host-based adsorption of the sera to
reduce non-specific cross-reactive antibody species. Monoclonal
antibodies avoid this pitfall but frequently result in reagents
whose specific epitopic specificity is unknown.
SUMMARY OF THE INVENTION
[0004] The invention relates to a method of identifying an
antigenic amino acid subsequence from within a larger amino acid
sequence comprising the steps of evaluating the hydrophilicity of
subsequences of an amino acid sequence of interest; evaluating the
flexibilitiy of subsequences of the amino acid sequence of
interest; and selecting an amino acid subsequence having
overlapping regions of hydrophilicity and flexibility. In
particular embodiments, the larger amino acid sequence is selected
from the group consisting of polypeptides expressed by members of
the Chlamydia genus.
[0005] The invention also relates to antigenic amino acid
subsequences identified by the methods described herein. In
particular embodiemnts, the invention pertains to an antigenic
amino acid subsequence selected from the group consisting of SEQ ID
NOS: 1-118.
[0006] The invention also pertains to antibodies which are specific
for the antigenic amino acid subsequences described herein. For
example, the invention pertains to monoclonal antibodies specific
for antigenic amino acid subsequences described herein.
[0007] The invention also relates to diagnostic and therapeutic
methods utilizing the described antigenic amino acid subsequences
and antibodies thereto.
BRIEF DESCRIPTION OF THE DRAWINGS
[0008] FIGS. 1A and 1B are sequence alignments of various Chlamydia
MOMPs. Variable domains (VD1-VD4) are boxed. Sequences are aligned
with the L2 serovar of C. trachomatis and are ranked from highest
homology (B, D, E, L1) to lower homology (F, C, and A, H, L3). MU
is the mouse pneumonitis C. trachomatis. PN refers to the human C.
pneumonia. Deletions are indicated by (-). A blank indicates the
same residue as L2. The leader sequence is bracketed. Underlined
seven residue segments are predicted to contain the most flexible
peptide backbone based on the L2 sequence. Asterisks indicate the
most hydrophilic region.
[0009] FIG. 2 illustrates the predicted antigenic sequences from
variable domains 1 (VD1) of various Chlamydia species. The boxed
cysteine (C) residue is not part of the native sequence but has
been added at the amino terminus for cross-linking to carrier
proteins used in immunization.
[0010] FIG. 3 illustrates the predicted antigenic sequences from
variable domain 2 (VD2) of various Chlamydia species. The boxed
cysteine (C) residue is not part of the native seuqence but has
been added at the amino terminus for cross-linking to carrier
proteins used in immunization.
[0011] FIG. 4 illustrates the predicted antigenic sequences from a
common domain of various Chlamydia species. The shaded box
indicates hydrophilic mobile region common to each with expected
cross-reactivity for antibodies specific for the sequence. The
boxed cysteine (C) residue is not part of the native sequence but
has been added at the amino terminus for cross-linking to carrier
proteins used in immunization.
DETAILED DESCRIPTION OF THE INVENTION
[0012] Globular proteins have a hydrophobic core, with the external
surfaces bearing relatively hydrophilic sequences. It is these
segments in native proteins which are most likely to be recognized
by antibodies. Work described herein describes methods for
identifying linear amino acid antigenic sequences for the
production of both polyclonal and monoclonal antibodies to defined
antigenic domains. One significant advantage of this technique is
that it provides antibodies to a known epitope of a target antigen
or organism.
[0013] The identification of antigenic domains described herein is
based on the overlap of the most hydrophilic peptide segments of an
antigen with those peptide segments with a concomitant predicted
peptide flexibility. Increased flexibility allows more
conformational degrees of freedom for optimal fit into an antibody
binding site. Aromatic amino acids are frequently found in
antigenic epitopes although hydrophobic with bulky R groups. This
decrease in the relative hydrophobicity and flexibility of th
peptide sequence containing the aromatic residue is compensated for
if accessible (i.e., surface of the antigen).
[0014] The relative hydrophilicity of peptide domains is based on
the individual hydrophilicity of each amino acid in six or more
residue segments as defined by Hopp and Woods (Hopp and Woods,
Proceedings of National Academy of Sciences USA 78:3824-3828).
Flexibility of the peptide chain at each C.alpha. residue is
measured from the average value of the atomic temperature factor as
affected by adjacent residues. Amino acids which result in rigidity
of the chain include alanine, valine, leucine, isoleucine,
tyrosine, phenylalanine, tryptophan, cysteine, methionine, and
histidine. Flexibility is computed by averaging the rigidity factor
(B value) along a seven residue segment using the following
expression (Karplus and Shulz, Naturwissenschaften 72:212-213
(1985); Van Regenmortel, Trends in Biochemical Sciences (TIBS)
12:36-39 (1986)):
F=B.sub.1+0.75(B.sub.i-1+B.sub.i+1)+0.5
(B.sub.1-2+B.sub.i+2)+0.25(B.sub.i- -3+B.sub.1+3)
[0015] With respect to identification of larger proteins or
polypeptides from which the antigenic amino acid subsequences are
selected, bands idenitifed by gel analysis can be isolated and
purified by HPLC, and the resulting purified protein can be
sequenced. Alternatively, the purified protein can be enzymatically
digested by methods known in the art to produce polypeptide
fragments which can be sequenced. The sequencing can be performed,
for example, by the methods of Wilm et al. (Nature
379(6564):466-469 (1996)). The protein can be isolated by
conventional means of protein biochemistry and purification to
obtain a substantially pure product, i.e., 80, 95 or 99% free of
cell component contaminants, as described in Jacoby, Methods in
Enzymology Volume 104, Academic Press, New York (1984); Scopes,
Protein Purification, Principles and Practice, 2nd Edition,
Springer-Verlag, New York (1987); and Deutscher (ed), Guide to
Protein Purification, Methods in Enzymology, Vol. 182 (1990). If
the protein is secreted, it can be isolated from the supernatant in
which the host cell is grown. If not secreted, the protein can be
isolated from a lysate of the host cells.
[0016] In addition to substantially full-length polypeptides used
as the source of the selected antigenic amino acid subsequences,
biologically active fragments of polypeptides, or analogs thereof,
including organic molecules which simulate the interactions of the
polypeptides, can be used. Biologically active fragments include
any portion of the full-length polypeptide which has a biological
function, including ligand binding, and antibody binding. Ligand
binding includes binding by nucleic acids, proteins or
polypeptides, small biologically active molecules, or large
cellular structures. Amino acid sequences identified as antigenic
from de novo sequence determination of cDNA reading frames or the
isolated protein of interest, or by established sequences from
GeneBank and the (PDB), can be most conveniently synthesized by
solid phase peptide synthesis using either standard F-Monc or t-Boc
methodologies.
[0017] This invention also pertains to an isolated polypeptide
comprising the antigenic amino acid subsequences of the invention.
The encoded proteins or polypeptides of the invention can be
partially or substantially purified (e.g., purified to
homogeneity), and/or are substantially free of other proteins.
According to the invention, the amino acid sequence of the
polypeptide can be that of the naturally-occurring polypeptide or
can comprise alterations therein. Such alterations include
conservative or non-conservative amino acid substitutions,
additions and deletions of one or more amino acids; however, such
alterations should preserve at least one activity of the encoded
protein or polypeptide, i.e., the altered or mutant protein should
be an active derivative of the naturally-occurring protein. For
example, the mutation(s) can preferably preserve the three
dimensional configuration of the binding and/or catalytic site of
the native protein, the hydrophilicity and/or flexibility of the
polypeptide. The presence or absence of biological activity or
activities can be determined by various functional assays as
described herein. Moreover, amino acids which are essential for
antigenicity or the function of the encoded protein or polypeptide
can be identified by methods known in the art. Particularly useful
methods include identification of conserved amino acids in the
family or subfamily, site-directed mutagenesis and alanine-scanning
mutagenesis (for example, Cunningham and Wells, Science
244:1081-1085 (1989)), crystallization and nuclear magnetic
resonance. The altered polypeptides produced by these methods can
be tested for particular biologic activities, including
immunogenicity and antigenicity, as described herein.
[0018] Specifically, appropriate amino acid alterations can be made
on the basis of several criteria, including hydrophobicity,
hydrophilicity, basic or acidic character, charge, polarity, size,
the presence or absence of a functional group (e.g., --SH or a
glycosylation site), rigidity or flexibility, and aromatic
character. Assignment of various amino acids to similar groups
based on the properties above will be readily apparent to the
skilled artisan; further appropriate amino acid changes can also be
found in Bowie et al. (Science 247:1306-1310 (1990)).
[0019] Polypeptides of the invention can also be a fusion protein
comprising all or a portion of the amino acid sequence fused to an
additional component. Additional components, such as radioisotopes
and antigenic tags, can be selected to assist in the isolation or
purification of the polypeptide or to extend the half life of the
polypeptide; for example, a hexahistidine tag would permit ready
purification by nickel chromatography. Polypeptides or amino acid
sequences described herein can be isolated from naturally-occurring
sources, chemically synthesized or recombinantly produced by
methods known in the art.
[0020] The present invention also relates to nucleotide sequences
(nucleic acid molecules) which encode the antigenic amino acid
subsequences or polypeptides of the invention. As appropriate,
nucleic acid molecules of the present invention can be RNA, for
example, mRNA, or DNA, such as cDNA and genomic DNA. DNA molecules
can be double-stranded or single-stranded; single stranded RNA or
DNA can be either the coding, or sense, strand or the non-coding,
or antisense, strand. The nucleic acid molecule can include all or
a portion of the coding sequence of a gene and can further comprise
additional non-coding sequences such as introns and non-coding 3'
and 5' sequences (including regulatory sequences, for example).
Additionally, the nucleic acid molecule can be fused to a marker
sequence, for example, a sequence which encodes a polypeptide to
assist in isolation or purification of the polypeptide. Such
sequences include, but are not limited to, those which encode a
glutathione-S-transferase (GST) fusion protein and those which
encode a hemaglutin A (HA) polypeptide marker from influenza.
[0021] As used herein, an "isolated" gene or nucleic acid molecule
is intended to mean a gene or nucleic acid molecule which is not
flanked by nucleic acid molecules which normally (in nature) flank
the gene or nucleic acid molecule (such as in genomic sequences)
and/or has been completely or partially purified from other
transcribed sequences (as in a cDNA or RNA library). For example,
an isolated nucleic acid of the invention may be substantially
isolated with respect to the complex cellular milieu in which it
naturally occurs. In some instances, the isolated material will
form part of a composition (for example, a crude extract containing
other substances), buffer system or reagent mix. In other
circumstance, the material may be purified to essential
homogeneity, for example as determined by PAGE or column
chromatography such as HPLC. Preferably, an isolated nucleic acid
comprises at least about 50, 80 or 90 percent (on a molar basis) of
all macromolecular species present. Thus, an isolated gene or
nucleic acid molecule can include a gene or nucleic acid molecule
which is synthesized chemically or by recombinant means.
Recombinant DNA contained in a vector are included in the
definition of "isolated" as used herein. Also, isolated nucleic
acid molecules include recombinant DNA molecules in heterologous
host cells, as well as partially or substantially purified DNA
molecules in solution. In vivo and in vitro RNA transcripts of the
DNA molecules of the present invention are also encompassed by
"isolated" nucleic acid molecules. Such isolated nucleic acid
molecules are useful in the manufacture of the encoded polypeptide,
as probes for isolating homologous sequences (e.g., from other
mammalian species), for gene mapping (e.g., by in situ
hybridization with chromosomes), or for detecting expression of the
gene in tissue (e.g., human tissue such as liver tissue), such as
by Northern blot analysis.
[0022] The invention also pertains to nucleic acid molecules which
hybridize under high stringency hybridization conditions (e.g., for
selective hybridization) to a nucleotide sequence described herein.
Hybridization probes are oligonucleotides which bind in a
base-specific manner to a complementary strand of nucleic acid.
Appropriate stringency conditions are known to those skilled in the
art or can be found in standard texts such as Current Protocols in
Molecular Biology, John Wiley & Sons, N.Y. (1989), 6.3.1-6.3.6.
For example, stringent hybridization conditions include a salt
concentration of no more than 1 M and a temperature of at least
25.degree. C. In one embodiment, conditions of 5.times.SSPE (750 mM
NaCl, 50 mM NaPhosphate, 5 mM EDTA, pH 7.4) and a temperature of
25-30.degree. C., or equivalent conditions, are suitable for
specific probe hybridizations. Equivalent conditions can be
determined by varying one or more of the parameters given as an
example, as known in the art, while maintaining a similar degree of
identity or similarity between the target nucleic acid molecule and
the primer or probe used. Hybridizable nucleic acid molecules are
useful as probes and primers for diagnostic applications.
[0023] Accordingly, the invention pertains to nucleic acid
molecules which have a substantial identity with the nucleic acid
molecules described herein and which encode antigenic amino acid
sequences; particularly preferred are nucleic acid molecules which
have at least about 90%, and more preferably at least about 95%
identity with nucleic acid molecules described herein. Thus, DNA
molecules which comprise a sequence which is different from the
naturally-occurring nucleic acid molecule but which, due to the
degeneracy of the genetic code, encode the same polypeptide are the
subject of this invention. The invention also encompasses
variations of the nucleic acid molecules of the invention, such as
those encoding portions, analogues or derivatives of the encoded
polypeptide. Such variations can be naturally-occurring, such as in
the case of allelic variation, or non-naturally-occurring, such as
those induced by various mutagens and mutagenic processes. Intended
variations include, but are not limited to, addition, deletion and
substitution of one or more nucleotides which can result in
conservative or non-conservative amino acid changes, including
additions and deletions. Preferably, the nucleotide or amino acid
variations are silent; that is, they do not alter the
characteristics or activity of the encoded protein or polypeptide.
As used herein, activities of the encoded protein or polypeptide
include, but are not limited to, catalytic activity, binding
function, antigenic function and oligomerization function.
[0024] The nucleotide sequences described herein can be amplified
as needed by methods known in the art. For example, this can be
accomplished by e.g., PCR. See generally PCR Technology: Principles
and Applications for DNA Amplification (ed. H. A. Erlich, Freeman
Press, NY, N.Y., 1992); PCR Protocols: A Guide to Methods and
Applications (eds. Innis, et al., Academic Press, San Diego,
Calif., 1990); Mattila et al., Nucleic Acids Res. 19, 4967 (1991);
Eckert et al., PCR Methods and Applications 1, 17 (1991); PCR (eds.
McPherson et al., IRL Press, Oxford); and U.S. Pat. No.
4,683,202.
[0025] Other suitable amplification methods include the ligase
chain reaction (LCR) (see Wu and Wallace, Genomics 4, 560 (1989),
Landegren et al., Science 241, 1077 (1988), transcription
amplification (Kwoh et al., Proc. Natl. Acad. Sci. USA 86, 1173
(1989)), and self-sustained sequence replication (Guatelli et al.,
Proc. Nat. Acad. Sci. USA, 87, 1874 (1990)) and nucleic acid based
sequence amplification (NASBA). The latter two amplification
methods involve isothermal reactions based on isothermal
transcription, which produce both single stranded RNA (ssRNA) and
double stranded DNA (dsDNA) as the amplification products in a
ratio of about 30 or 100 to 1, respectively.
[0026] The amplified DNA can be radiolabelled and used as a probe
for screening a cDNA library. Corresponding clones can be isolated,
DNA can obtained following in vivo excision, and the cloned insert
can be sequenced in either or both orientations by art recognized
methods, to identify the correct reading frame encoding a protein
of the appropriate molecular weight. For example, the direct
analysis of the nucleotide sequence of nucleic acid molecules of
the present invention can be accomplished using either the dideoxy
chain termination method or the Maxam Gilbert method (see Sambrook
et al., Molecular Cloning, A Laboratory Manual (2nd Ed., CSHP, New
York 1989); Zyskind et al., Recombinant DNA Laboratory Manual,
(Acad. Press, 1988)). Using these or similar methods, the
polypeptide(s) and the DNA encoding the polypeptide can be
isolated, sequenced and further characterized.
[0027] The invention also provides expression vectors containing a
nucleic acid sequence described herein, operably linked to at least
one regulatory sequence. Many such vectors are commercially
available, and other suitable vectors can be readily prepared by
the skilled artisan. "Operably linked" is intended to mean that the
nucleic acid molecule is linked to a regulatory sequence in a
manner which allows expression of the nucleic acid sequence.
Regulatory sequences are art-recognized and are selected to produce
the encoded polypeptide. Accordingly, the term "regulatory
sequence" includes promoters, enhancers, and other expression
control elements which are described in Goeddel, Gene Expression
Technology: Methods in Enzymology 185, Academic Press, San Diego,
Calif. (1990). For example, the native regulatory sequences or
regulatory sequences native to the transformed host cell can be
employed. It should be understood that the design of the expression
vector may depend on such factors as the choice of the host cell to
be transformed and/or the type of polypeptide desired to be
expressed. For instance, the polypeptides of the present invention
can be produced by ligating the cloned gene, or a portion thereof,
into a vector suitable for expression in either prokaryotic cells,
eukaryotic cells or both (see, for example, Broach, et al.,
Experimental Manipulation of Gene Expression, ed. M. Inouye
(Academic Press, 1983) p. 83; Molecular Cloning: A Laboratory
Manual, 2nd Ed., ed. Sambrook et al. (Cold Spring Harbor Laboratory
Press, 1989) Chapters 16 and 17). Typically, expression constructs
will contain one or more selectable markers, including, but not
limited to, the gene that encodes dihydrofolate reductase and the
genes that confer resistance to neomycin, tetracycline, ampicillin,
chloramphenicol, kanamycin and streptomycin resistance.
[0028] Prokaryotic and eukaryotic host cells transfected by the
described vectors are also provided by this invention. For
instance, cells which can be transfected with the vectors of the
present invention include, but are not limited to, bacterial cells
such as E. coli (e.g., E. coli K12 strains, Streptomyces,
Pseudomonas, Serratia marcescens and Salmonella typhimurium, insect
cells (baculovirus), including Drosophila, fungal cells, such as
yeast cells, plant cells and mammalian cells, such as thymocytes,
Chinese hamster ovary cells (CHO), and COS cells.
[0029] Thus, a nucleic acid molecule described herein can be used
to produce a recombinant form of the polypeptide via microbial or
eukaryotic cellular processes. Ligating the polynucleic acid
molecule into a gene construct, such as an expression vector, and
transforming or transfecting into hosts, either eukaryotic (yeast,
avian, insect, plant or mammalian) or prokaryotic (bacterial
cells), are standard procedures used in producing other well known
proteins. Similar procedures, or modifications thereof, can be
employed to prepare recombinant polypeptides according to the
present invention by microbial means or tissue-culture technology.
Accordingly, the invention pertains to the production of encoded
polypeptides by recombinant technology.
[0030] The polypeptides of the present invention can be isolated or
purified (e.g., to homogeneity) from recombinant cell culture by a
variety of processes. These include, but are not limited to, anion
or cation exchange chromatography, ethanol precipitation, affinity
chromatography and high performance liquid chromatography (HPLC).
The particular method used will depend upon the properties of the
polypeptide and the selection of the host cell; appropriate methods
will be readily apparent to those skilled in the art.
[0031] The present invention also relates to antibodies which bind
an antigenic amino acid sequence or subsequence of the invention.
For instance, polyclonal and monoclonal antibodies, including
non-human and human antibodies, humanized antibodies, chimeric
antibodies and antigen-binding fragments thereof (Current Protocols
in Immunology, John Wiley & Sons, N.Y. (1994); EP Application
173,494 (Morrison); International Patent Application WO86/01533
(Neuberger); and U.S. Pat. No. 5,225,539 (Winters)) which bind to
the described amino acid sequence or subsequence are within the
scope of the invention. A mammal, such as a mouse, rat, hamster or
rabbit, can be immunized with an immunogenic form of the amino acid
subsequence. Techniques for conferring immunogenicity on a
polypeptide include conjugation to carriers or other techniques
well known in the art. The polypeptide can be administered in the
presence of an adjuvant. The progress of immunization can be
monitored by detection of antibody titers in plasma or serum.
Standard ELISA or other immunoassays can be used with the immunogen
as antigen to assess the levels of antibody.
[0032] Following immunization, anti-peptide antisera can be
obtained, and if desired, polyclonal antibodies can be isolated
from the serum. Monoclonal antibodies can also be produced by
standard techniques which are well known in the art (Kohler and
Milstein, Nature 256:495-497 (1975); Kozbar et al., Immunology
Today 4:72 (1983); and Cole et al., Monoclonal Antibodies and
Cancer Therapy, Alan R. Liss, Inc., pp. 77-96 (1985)). The term
"antibody" as used herein is intended to include fragments thereof,
such as Fab and F(ab).sub.2 and antigen binding fragments.
Antibodies described herein can be used to inhibit the activity of
the polypeptides and proteins described herein, particularly in
vitro and in cell extracts, using methods known in the art.
[0033] Additionally, such antibodies, in conjunction with a label,
such as a radioactive label, can be used to assay for the presence
of the expressed protein in a cell from, e.g., a tissue sample, and
can be used in an immunoabsorption process, such as an ELISA, to
isolate the protein or polypeptide. Tissue samples which can be
assayed include human tissues, e.g., differentiated and
non-differentiated cells. Examples include bone marrow, thymus,
kidney, liver, brain, pancreas, fibroblasts and epithelium. These
antibodies are useful in diagnostic assays, or as an active
ingredient in a pharmaceutical composition.
[0034] The invention also relates to immunogenic compositions
comprising amino acid sequences described herein, as well as
vaccine compositions comprising polypeptides or antibodies
described herein. Peptides and antibodies identified by methods
described herein can also be used in a variety of assay and protein
processing applications, including, but not limited to,
radioimmunoassays, ELISA, antigen capture assays, competitive
inhibition assays, affinity chromatography, Western Blotting,
Labeled-antibody assays such as immunoflorescence assays,
immunohistochemical staining assays and immunoprecipitation assays.
The antiobdies, alone or linked to particular toxins, can also be
used for a variety of therapeutic and other purposes, inclduing
removing specific lymphocyte subsets, inhibiting cell function,
inhibiting graft rejection, alleviating or suppressing autoimmune
disease, and attaching to tumors.
[0035] The present invention also pertains to pharmaceutical
compositions comprising antigenic amino acid sequences or
subsequences and other antibodies described herein. For instance, a
composition of the present invention can be formulated with a
physiologically acceptable medium to prepare a pharmaceutical
composition. The particular physiological medium may include, but
is not limited to, water, buffered saline, polyols (e.g., glycerol,
propylene glycol, liquid polyethylene glycol) and dextrose
solutions. The optimum concentration of the active ingredient(s) in
the chosen medium can be determined empirically, according to well
known procedures, and will depend on the ultimate pharmaceutical
formulation desired. Methods of introduction of exogenous
polypeptides at the site of treatment include, but are not limited
to, intradermal, intramuscular, intraperitoneal, intravenous,
subcutaneous, oral and intranasal. Other suitable methods of
introduction can also include gene therapy, rechargeable or
biodegradable devices and slow release polymeric devices. The
pharmaceutical compositions of this invention can also be
administered as part of a combinatorial therapy with other
agents.
[0036] The nucleic acid sequences described herein can also be used
for genetic immunization. The term, "genetic immunization", as used
herein, refers to inoculation of a vertebrate, particularly a
mammal, with a nucleic acid vaccine directed against a pathogenic
agent, such as Chlamydia, resulting in protection of the vertebrate
against the pathogenic agent. Representative vertebrates include
mice, dogs, cats, chickens, sheep, goats, cows, horses, pigs,
non-human primates, and humans. A "nucleic acid vaccine" or "DNA
vaccine" as used herein, is a nucleic acid construct comprising a
polynucleotide encoding a polypeptide antigen, particularly an
antigenic amino acid subsequence identified by methods described
herein. The nucleic acid construct can also include transcriptional
promoter elements, enhancer elements, splicing signals, termination
and polyadenylation signals, and other nucleic acid sequences.
[0037] "Protection against the pathogenic agent" as used herein
refers to generation of an immune response in the vertebrate, the
immune response being protective (partially or totally) against
manifestations of the disease caused by the pathogenic agent. A
vertebrate that is protected against disease may be infected with
the pathogenic agent, but to a lesser degree than would occur
without immunization; may be infected with the pathogenic agent,
but does not exhibit disease symptoms; or may be infected with the
pathogenic agent, but exhibits fewer disease symptoms than would
occur without immunization. Alternatively, the vertebrate that is
protected against disease may not become infected with the
pathogenic agent at all, despite exposure to the agent.
[0038] The nucleic acid vaccine can be produced by standard
methods. For example, using known methods, a nucleic acid encoding
polypeptide antigen of interest, e.g., DNA encoding an antigenic
amino acid subsequence, can be inserted into an expression vector
to construct a nucleic acid vaccine (see Maniatis et al., Molecular
Cloning, A Laboratory Manual, 2nd edition, Cold Spring Harbor
Laboratory Press (1989)).
[0039] The individual vertebrate is inoculated with the nucleic
acid vaccine (i.e., the nucleic acid vaccine is administered),
using standard methods. The vertebrate can be inoculated
subcutaneously, intravenously, intraperitoneally, intradermally,
intramuscularly, topically, orally, rectally, nasally, buccally,
vaginally, by inhalation spray, or via an implanted reservoir in
dosage formulations containing conventional non-toxic,
physiologically acceptable carriers or vehicles. Alternatively, in
a preferred embodiment, the vertebrate is innoculated with the
nucleic acid vaccine through the use of a particle acceleration
instrument (a "gene gun"). The form in which it is administered
(e.g., capsule, tablet, solution, emulsion) will depend in part on
the route by which it is administered. For example, for mucosal
administration, nose drops, inhalants or suppositories can be
used.
[0040] The nucleic acid vaccine can be administered in conjunction
with known adjuvants. The adjuvant is administered in a sufficient
amount, which is that amount that is sufficient to generate an
enhanced immune response to the nucleic acid vaccine. The adjuvant
can be administered prior to (e.g., 1 or more days before)
inoculation with the nucleic acid vaccine; concurrently with (e.g.,
within 24 hours of) inoculation with the nucleic acid vaccine;
contemporaneously (simultaneously) with the nucleic acid vaccine
(e.g., the adjuvant is mixed with the nucleic acid vaccine, and the
mixture is administered to the vertebrate); or after (e.g., 1 or
more days after) inoculation with the nucleic acid vaccine. The
adjuvant can also be administered at more than one time (e.g.,
prior to inoculation with the nucleic acid vaccine and also after
inoculation with the nucleic acid vaccine). As used herein, the
term "in conjunction with" encompasses any time period, including
those specifically described herein and combinations of the time
periods specifically described herein, during which the adjuvant
can be administered so as to generate an enhanced immune response
to the nucleic acid vaccine (e.g., an increased antibody titer to
the antigen encoded by the nucleic acid vaccine, or an increased
antibody titer to the pathogenic agent). The adjuvant and the
nucleic acid vaccine can be administered at approximately the same
location on the vertebrate; for example, both the adjuvant and the
nucleic acid vaccine are administered at a marked site on a limb of
the vertebrate.
[0041] In a particular embodiment, the nucleic acid construct is
co-administered with a transfection-facilitating cationic lipid. In
a preferred embodiment, the cationic lipid is dioctylglycylspermine
(DOGS) (U.S. patent application Ser. Nos. 08/372,429 and
08/544,575, PCT application Ser. No. PCT/US96/16845 and published
PCT application publication no. WO 96/21356). In a particular
embodiment, the nucleic acid construct is co-administered with a
transfection-facilitating cationic lipid and an amount of
1,25(OH).sub.2D3 effective to produce a mucosal response. In a
preferred embodiment, the nucleic acid construct is complexed with
a transfection-facilitating cationic lipid.
[0042] The teachings of all references cited herein are are
spceifically incorporated herein by reference. The teachings of
Attorney Docket No. VDB96-02pA2, entitled "Diagnosis and Management
of Infection Caused by Chlamydia" by William M. Mitchell and
Charles W. Stratton, filed concurrently with the present
application, are also incorporated herein by reference in their
entirety.
EXAMPLES
[0043] Examples of the predictive power of the methodology
described herein include the following:
[0044] 1) Antigenicity of the MOMP (Major Outer Membrane Protein)
of Chlamydia:
[0045] In order to provide ELISA assays that are species- and
potentially strain-specific for the various Chlamydia, two regions
in the MOMP have been identified which show minimal amino acid
sequence homologies and which are predicted to be excellent
antigenic domains by virtue of hydrophilicity and peptide mobility
on the solvent-accessible surface of MOMP. FIG. 1 illustrates the
constant and variable domain (VD) of the various chlamydial
species. The identified species-specific antigenic domains are
located in VD1 and VD2. FIG. 2 illustrates the peptide amino acid
sequences employed for the construction of peptide based ELISAs
with species specificity for VD1. FIG. 3 illustrates the peptides
for VD2 which are used similarly to the VD1 sequences. In addition,
a highly antigenic domain (FIG. 4) common to all Chlamydia has been
identified and developed as genus-specific ELISA for the Chlamydia.
Immunization of rabbits has verified the antigenicity of each
peptide to each peptide (Table 1). Monoclonal antibodies have
further verified the specificities and antigenicity of each peptide
(Table 1) as predicted by computer analysis of the
nucleotide-generated amino acid sequence of each species-specific
MOMP.
[0046] Table 1: Antigenic Responses To Peptides From Four Species
Of Chlamydiae Identified By Hydrophilicity And Peptide Movement As
Highly Antigenic
1 Chlamydiae Titer.sup.a Species Peptide.sup.b Pre Post C.
pneumoniae 90-105 100 >3200 C. trachomatis L2 91-106 800
>3200 C. psittaci 92-106 400 >3200 C. trachomatis (mouse)
89-105 0 >3200 C. pneumoniae 158-171 25 >3200 C. trachomatis
L2 159-175 200 >3200 C. psittaci 160-172 100 >3200 C.
trachomatis (mouse) 158-171 800 >3200 C. pneumoniae 342-354 200
>3200 C. trachomatis L2 342-354 100 >3200 C. psittaci
ND.sup.c C. trachomatis (mouse) ND.sup.c .sup.aReciprocal titer
.sup.bImmunogenic peptide and ELISA antigen of specific amino acid
sequence against the indicated pre-immunization and
post-immunization rabbit serum .sup.cND, not done
[0047] Table 2 illustrates reciprocal titers of a polyclonal and
monoclonal antibody against C. trachomatis cross-reactive against a
C. pneumoniae peptide encompassing amino acids 342-354 and a
recombinant full length MOMP from C. pneumoniae. Note that the
monoclonal antibody raised against C. trachomatis has as its
epitope genus-specific reactivity against peptide 342-354 of C.
pneumoniae.
2 TABLE 2 Titer.sup.a Antigen Polyclonal Ab.sup.b Monoclonal
Ab.sup.c CPN Momp.sup.d 400 0 CPN 90-105.sup.e 50 0 CPN
158-171.sup.f 50 0 CPN 342-354.sup.g >3200 1600 .sup.aReciprocal
titer .sup.bPolyclonal goat Ab from Chemicon International, Inc.
(Temecula, CA) against MOMP of C. trachomatis .sup.CMonoclonal Ab
from ICN Immunologicals (Costa Mesa, CA) against MOMP of C.
trachomatis .sup.dC. pneumoniaerecombinant MOMP .sup.eAmino acid
peptide 90-105 of C. pneumoniae .sup.fAmino acid peptide 158-171 of
C. pneumoniae .sup.gAmino acid peptide 342-354 of C. pneumoniae
[0048] 2) Antigenicity of the 76 k D Protein of C. Pneumoniae:
[0049] C. pneumoniae expresses a gene encoding a unique 76 kD
protein (Perez-Melgosa et al., Infect. Immun. 62:880-886 (1994)).
Hydrophilicity/peptide flexibility analysis predicts the sequence
of amino acids 302-315 (KPKESKTDSVERWS; SEQ ID NO: 1) to be highly
antigenic; the peptide has been extended towards the carboxyl
terminus to include aromatic and additional hydrophilic amino acid
residues. The predicted sequence has been further modified to
include an adjacent relatively hydrophilic region containing an
aromatic amino acid (tryptophan). Other potential antigenic
peptides based on either hydrophilicity or peptide flexibility and
extended to include emino acids found in hydrophilic or flexible
segments, as well as inclusion of aromatic amino acids immediately
adjacent to the predicted antigens, are illustrated in Table 3.
3TABLE 3 a) Peptide Movement Predictions SSNSSSSTSRS (SEQ ID NO: 2)
AA 335-345 GSKQQGSS (SEQ ID NO: 3) AA 599-606 GKAGQQQG (SEQ ID NO:
4) AA 683-690 PSETSTTEK (SEQ ID NO: 5) AA 35-43 KPADGSDV (SEQ ID
NO: 6) AA 583-590 NGQKKPLYLYG (SEQ ID NO: 7) AA 70-80
SDVPNPGTTVGGSKQQGSS (SEQ ID NO: 8) AA 588-606 HMFNTENPDSQAAQQ (SEQ
ID NO: 9) AA 636-650 b) Hydrophilic Prediction DDAENETAS (SEQ ID
NO: 10) AA 617-625
[0050] 3) Antigenicity of the Chlamydial Heat Shock Proteins:
[0051] C. pneumoniae expresses three known genes with significant
homology to the human heat shock proteins of 70, 60 and 10 kD.
Antigenicity of homologous regions may result in molecular mimicry
and autoimmunity. Indeed, it is postulated that the tubal scarring
secondary to infection from C. trachomatis is due to cross-reactive
cell mediated immunity against one or more heat shock proteins.
[0052] a) C. pneumonia DNAK/Heatshock Protein 70:
[0053] Hydrophilicity/peptide flexibility analysis predicts a
highly antigenic sequence in the C-terminal region of the expressed
protein. This antigenic domain and its homologous human protein are
illustrated in Table 4; vertical lines indicate residue homology
while "+" signs indicate retention of a positive charge at the
site. Amino acid residues 522-529 are either homologous to the
human protein or possess preservation of charge (i.e., AA 525-529).
Antibodies against this epitope would be expected to possess
cross-reactivity with the human 70 kD heat shock protein. Peptides
incorporating the C-terminal end of this common region with the
non-homologous sequence would be expected to identify
Chlamydial-specific antibodies. Two embodiments of this invention
include the full length peptide (AA 521-536) and the
Chlamydial-specific epitopic sequence identified as AA 527-536 or
truncated for the identification of Chlamydia-specific antibodies.
Table 5 illustrates other potential antigenic sequences for the
DNAK protein expressed by C. pneumoniae based on either peptide
flexibility or hydrophilicity and extended to include amino acids
found in adjacent hydrophilic or flexible segments, as well as
inclusion of aromatic amino acids immediately adjacent to the
predicted antigens.
4TABLE 4 C. pneumoniae KEEDKKRREASDAKNE (SEQ ID NO: 11) (AA
521-536) .vertline..vertline..vertline.++++.vertline. .vertline.
human hsp70 AEEDRRKKERVEAVNM (SEQ ID NO: 12) (AA 569-584)
[0054]
5TABLE 5 KKHSFSTKPPSNNGSSEDHIEE (SEQ ID NO: 13) (AA 628- 649)
YTVTSGSKGDAVFE (SEQ ID NO: 14) (AA 94-107) TSSEGTRTTPS (SEQ ID NO:
15) (AA 34-44) SEHKKSSK (SEQ ID NO: 16) (AA 2-9) KDVASGKEQKIRIE
(SEQ ID NO: 17) (AA 487- 500) ERNTTIPTQKKQIFST (SEQ ID NO: 18) (AA
411- 426) YFNDSQRASSTKDAGR (SEQ ID NO: 19) (AA 148- 162)
EEFKKQEGIDLSKDN (SEQ ID NO: 20) (AA 240- 254) NAKGGPNINTED (SEQ ID
NO: 21) (AA 615- 626) GERPMAKDNKETGRFD (SEQ ID NO: 22) (AA 441-
456)
[0055] b) C. pneumoniae GROEL/Heatshock Protein (hsp 60) 60:
[0056] Two peptides expressed by the GROEL gene of C. pneumoniae
have a high correlation of hydrophilicity and segment mobility
(Table 6). Residues with similar negative charges are identified by
"*" symbols. The sequences are highly conserved between C.
pneumoniae heat shock protein (hsp) 60 and the human hsp 60
associated with the mitochondrion. Thus the potential for molecular
mimicry is high and is a likely site for the development of humoral
autoimmune responses. Other potential antigenic regions based on
either peptide flexibility or hydrophilicity and extended to
include amino acids found in adjacent hydrophilic or flexible
peptide segments, as well as inclusion of aromatic amino acids
immediately adjacent to the predicted areas, are illustrated in
Table 7.
6TABLE 6 C. pneumoniae TEIEMKEKKDRVDD (SEQ ID NO: 23) hsp 60 *
.vertline. .vertline..vertline..vertline..vertline- ..vertline.
.vertline. (AA 385-398) SDVEVNEKKDRVTD (SEQ ID NO: 24) human hsp 60
(AA 410-423) C. pneumoniae hsp 60 EDSTSDYDKEK (SEQ ID NO: 25) hsp
60 * .vertline..vertline.*.ver-
tline.*.vertline..vertline..vertline. (AA 354-364) DVTTSEYEKEK (SEQ
ID NO: 26) human hsp 60 (AA 410-420)
[0057]
7TABLE 7 DDKSSSA (SEQ ID NO: 27) (AA 528-534) KKQIEDSTSDYVSEE (SEQ
ID NO: 28) (AA 350-364) SSYFSTNPETQE (SEQ ID NO: 29) (AA 201-212)
EKVGKNGSITVEEADK (SEQ ID NO: 30) (AA 167-182) SKTADKAGDGTTTAT (SEQ
ID NO: 31) (AA 79-93)
[0058] c) C. pneumoniae GROES/Heat Shock Protein 10 (hsp 10):
[0059] Three peptides are highly correlated with respect to
hydrophilicity/peptide movement analysis. Comparison to mouse
chaperonin 10 indicates little homology of these bacterial
antigenic domains with C. pneumoniae hsp 10 (Table 8).
8TABLE 8 C. pneumoniae KREEEEATAR (SEQ ID NO: 32) (AA 20-29)
.vertline. .vertline. + mouse chaperonin 10 ERSAAETVTK (SEQ ID NO:
33) (AA 19-28) C. pneumoniae DTAKKKQDRAE (SEQ ID NO: 34) (AA 36-46)
* .vertline. .vertline. mouse chaperonin 10 EKSQGKVLQAT (SEQ ID NO:
35) (AA 35-45) C. pneumoniae GTGKRTDDGT (SEQ ID NO: 36) (AA 51-60)
.vertline. .vertline. + .vertline. mouse chaperonin 10 GSGGKGKSGE
(SEQ ID NO: 37) (AA 50-59)
[0060] 4) Antigenicity of the Crysteine-Rich Proteins of C.
pneumoniae
[0061] a) 60 kD/OMP B:
[0062] The second most abundant protein of the external matrix is a
60 kD protein containing 34 cysteines (6.1%). Table 9 illustrates
the single peptide domain with overlapping hydrophilicity and
peptide flexibility profiles. The sequence has been extended
towards the C-terminus to include additional hydrophilic amino
acids and two aromatic residues.
[0063] Table 10 illustrates several additional peptides with
potential antigenic profiles based on either peptide flexibility or
hydrophilicity and extended to include amino acids found in
adjacent hydrophilic or flexible peptide segments as well as
inclusion of aromatic acids immediately adjacent to the predicted
areas..
9TABLE 9 RRNKQPVEQKSRGAFCDKEFYPCEE (SEQ ID NO: 38) (AA 60- 84)
[0064]
10TABLE 10 DMRPGDKKVFTVEFCPQRR (SEQ ID NO: 39) (AA 278-296)
SSDPETTPTSDGKVWKIDR (SEQ ID NO: 40) (AA 157-176)
TSESNCGTCTSCAETTTHWK (SEQ ID NO: 41) (AA 418-437) KLGSKESVEFS (SEQ
ID NO: 42) (AA 511-521) TVYRICVTNRGSAEDT (SEQ ID NO: 43) (AA
459-474) EYSISVSNPGD (SEQ ID NO: 44) (AA 343-353)
[0065] b) 9 kD Protein:
[0066] This small protein contains 14 cysteines (15.5%). Table 11
illustrates the predicted antigenic sites. Peptide 1 represents the
single peptide for the 9 kD cysteine-rich protein identified by
common hydrophilic/peptide flexibility profiles. Peptide 2
recognized initially by its peptide flexibility and extended
towards the amino terminal to include several hydrophilic
residues.
11TABLE 11 Peptide 1: RKKERS (SEQ ID NO: 105) (AA 44-49) Peptide 2:
STECNSQSPQ (SEQ ID NO: 106) (AA 68-77)
[0067] 5) Antigenicity of the Ebola Virus GP Protein:
[0068] The GP protein associates into trimers on the surface of the
virus and functions as an attachment protein. Two peptides are
predicted to be excellent antigens on the basis of overlapping
hydrophilic/peptide flexibility profiles (Table 12). Additional
potential antigenic sites initially based on either peptide
flexibility or hydrophilicity and extended to include amino acids
found in adjacent hydrophilic or flexible peptide segments as well
as inclusion of aromatic amino acids immediately adjacent to the
predicted domains are illustrated in Table 13.
12TABLE 12 NPNLHYWTTQDEG (SEQ ID NO: 107) (AA 512- 524)
SGQSPARTSSDPGTNTTTEDHK (SEQ ID NO: 108) (AA 320- 340)
[0069]
13TABLE 13 TGGRRTRRE (SEQ ID NO: 109) (AA 494-502) RDRFKRTSFF (SEQ
ID NO: 110) (AA 11-21) EQHHRRTDNDST (SEQ ID NO: 111) (AA 405-416)
ENTNTSKSTDF (SEQ ID NO: 112) (AA 433-443) YTSGKRSNTTGK (SEQ ID NO:
113) (AA 261-272) TTTSPQNHSET (SEQ ID NO: 114) (AA 448-458)
PDQGDNDNWWT (SEQ ID NO: 115) (AA 636-646) TISTSPQSLTTK (SEQ ID NO:
116) (AA 370-381) TEDPSSGYYSTTIRYQ (SEQ ID NO: 117) (AA 206-221)
THHQDTGEESASSGK (SEQ ID NO: 118) (AA 464-478)
[0070] Equivalents
[0071] Those skilled in the art will recognize, or be able to
ascertain, using no more than routine experimentation many
equivalents to the specific embodiments of the invention described
herein. Such equivalents are intended to be encompassed by the
following claims:
Sequence CWU 1
1
118 1 14 PRT Chlamydia pneumoniae 1 Lys Pro Lys Glu Ser Lys Thr Asp
Ser Val Glu Arg Trp Ser 1 5 10 2 11 PRT Chlamydia pneumoniae 2 Ser
Ser Asn Ser Ser Ser Ser Thr Ser Arg Ser 1 5 10 3 8 PRT Chlamydia
pneumoniae 3 Gly Ser Lys Gln Gln Gly Ser Ser 1 5 4 8 PRT Chlamydia
pneumoniae 4 Gly Lys Ala Gly Gln Gln Gln Gly 1 5 5 9 PRT Chlamydia
pneumoniae 5 Pro Ser Glu Thr Ser Thr Thr Glu Lys 1 5 6 8 PRT
Chlamydia pneumoniae 6 Lys Pro Ala Asp Gly Ser Asp Val 1 5 7 11 PRT
Chlamydia pneumoniae 7 Asn Gly Gln Lys Lys Pro Leu Tyr Leu Tyr Gly
1 5 10 8 19 PRT Chlamydia pneumoniae 8 Ser Asp Val Pro Asn Pro Gly
Thr Thr Val Gly Gly Ser Lys Gln Gln 1 5 10 15 Gly Ser Ser 9 15 PRT
Chlamydia pneumoniae 9 His Met Phe Asn Thr Glu Asn Pro Asp Ser Gln
Ala Ala Gln Gln 1 5 10 15 10 9 PRT Chlamydia pneumoniae 10 Asp Asp
Ala Glu Asn Glu Thr Ala Ser 1 5 11 16 PRT Chlamydia pneumoniae 11
Lys Glu Glu Asp Lys Lys Arg Arg Glu Ala Ser Asp Ala Lys Asn Glu 1 5
10 15 12 16 PRT Homo sapiens 12 Ala Glu Glu Asp Arg Arg Lys Lys Glu
Arg Val Glu Ala Val Asn Met 1 5 10 15 13 22 PRT Chlamydia
pneumoniae 13 Lys Lys His Ser Phe Ser Thr Lys Pro Pro Ser Asn Asn
Gly Ser Ser 1 5 10 15 Glu Asp His Ile Glu Glu 20 14 14 PRT
Chlamydia pneumoniae 14 Tyr Thr Val Thr Ser Gly Ser Lys Gly Asp Ala
Val Phe Glu 1 5 10 15 11 PRT Chlamydia pneumoniae 15 Thr Ser Ser
Glu Gly Thr Arg Thr Thr Pro Ser 1 5 10 16 8 PRT Chlamydia
pneumoniae 16 Ser Glu His Lys Lys Ser Ser Lys 1 5 17 14 PRT
Chlamydia pneumoniae 17 Lys Asp Val Ala Ser Gly Lys Glu Gln Lys Ile
Arg Ile Glu 1 5 10 18 16 PRT Chlamydia pneumoniae 18 Glu Arg Asn
Thr Thr Ile Pro Thr Gln Lys Lys Gln Ile Phe Ser Thr 1 5 10 15 19 16
PRT Chlamydia pneumoniae 19 Tyr Phe Asn Asp Ser Gln Arg Ala Ser Ser
Thr Lys Asp Ala Gly Arg 1 5 10 15 20 15 PRT Chlamydia pneumoniae 20
Glu Glu Phe Lys Lys Gln Glu Gly Ile Asp Leu Ser Lys Asp Asn 1 5 10
15 21 12 PRT Chlamydia pneumoniae 21 Asn Ala Lys Gly Gly Pro Asn
Ile Asn Thr Glu Asp 1 5 10 22 16 PRT Chlamydia pneumoniae 22 Gly
Glu Arg Pro Met Ala Lys Asp Asn Lys Glu Ile Gly Arg Phe Asp 1 5 10
15 23 14 PRT Chlamydia pneumoniae 23 Thr Glu Ile Glu Met Lys Glu
Lys Lys Asp Arg Val Asp Asp 1 5 10 24 14 PRT Homo sapiens 24 Ser
Asp Val Glu Val Asn Glu Lys Lys Asp Arg Val Thr Asp 1 5 10 25 11
PRT Chlamydia pneumoniae 25 Glu Asp Ser Thr Ser Asp Tyr Asp Lys Glu
Lys 1 5 10 26 11 PRT Homo sapiens 26 Asp Val Thr Thr Ser Glu Tyr
Glu Lys Glu Lys 1 5 10 27 7 PRT Chlamydia pneumoniae 27 Asp Asp Lys
Ser Ser Ser Ala 1 5 28 15 PRT Chlamydia pneumoniae 28 Lys Lys Gln
Ile Glu Asp Ser Thr Ser Asp Tyr Val Ser Glu Glu 1 5 10 15 29 12 PRT
Chlamydia pneumoniae 29 Ser Ser Tyr Phe Ser Thr Asn Pro Glu Thr Gln
Glu 1 5 10 30 16 PRT Chlamydia pneumoniae 30 Glu Lys Val Gly Lys
Asn Gly Ser Ile Thr Val Glu Glu Ala Asp Lys 1 5 10 15 31 15 PRT
Chlamydia pneumoniae 31 Ser Lys Thr Ala Asp Lys Ala Gly Asp Gly Thr
Thr Thr Ala Thr 1 5 10 15 32 10 PRT Chlamydia pneumoniae 32 Lys Arg
Glu Glu Glu Glu Ala Thr Ala Arg 1 5 10 33 10 PRT Mouse 33 Glu Arg
Ser Ala Ala Glu Thr Val Thr Lys 1 5 10 34 11 PRT Chlamydia
pneumoniae 34 Asp Thr Ala Lys Lys Lys Gln Asp Arg Ala Glu 1 5 10 35
11 PRT Mouse 35 Glu Lys Ser Gln Gly Lys Val Leu Gln Ala Thr 1 5 10
36 10 PRT Chlamydia pneumoniae 36 Gly Thr Gly Lys Arg Thr Asp Asp
Gly Thr 1 5 10 37 10 PRT Mouse 37 Gly Ser Gly Gly Lys Gly Lys Ser
Gly Glu 1 5 10 38 25 PRT Chlamydia pneumoniae 38 Arg Arg Asn Lys
Gln Pro Val Glu Gln Lys Ser Arg Gly Ala Phe Cys 1 5 10 15 Asp Lys
Glu Phe Tyr Pro Cys Glu Glu 20 25 39 19 PRT Chlamydia pneumoniae 39
Asp Met Arg Pro Gly Asp Lys Lys Val Phe Thr Val Glu Phe Cys Pro 1 5
10 15 Gln Arg Arg 40 19 PRT Chlamydia pneumoniae 40 Ser Ser Asp Pro
Glu Thr Thr Pro Thr Ser Asp Gly Lys Val Trp Lys 1 5 10 15 Ile Asp
Arg 41 20 PRT Chlamydia pneumoniae 41 Thr Ser Glu Ser Asn Cys Gly
Thr Cys Thr Ser Cys Ala Glu Thr Thr 1 5 10 15 Thr His Trp Lys 20 42
11 PRT Chlamydia pneumoniae 42 Lys Leu Gly Ser Lys Glu Ser Val Glu
Phe Ser 1 5 10 43 16 PRT Chlamydia pneumoniae 43 Thr Val Tyr Arg
Ile Cys Val Thr Asn Arg Gly Ser Ala Glu Asp Thr 1 5 10 15 44 11 PRT
Chlamydia pneumoniae 44 Glu Tyr Ser Ile Ser Val Ser Asn Pro Gly Asp
1 5 10 45 100 PRT Chlamydia trachomatis 45 Met Lys Lys Leu Leu Lys
Ser Val Leu Val Phe Ala Ala Leu Ser Ser 1 5 10 15 Ala Ser Ser Leu
Gln Ala Leu Pro Val Gly Asn Pro Ala Glu Pro Ser 20 25 30 Leu Met
Ile Asp Gly Ile Leu Trp Glu Gly Phe Gly Gly Asp Pro Cys 35 40 45
Asp Pro Cys Thr Thr Trp Cys Asp Ala Ile Ser Met Arg Met Gly Tyr 50
55 60 Tyr Gly Asp Phe Val Phe Asp Arg Val Leu Gln Thr Asp Val Asn
Lys 65 70 75 80 Glu Phe Gln Met Gly Ala Lys Pro Thr Thr Ala Thr Gly
Asn Ala Ala 85 90 95 Ala Pro Ser Thr 100 46 100 PRT Chlamydia
trachomatis 46 Met Lys Lys Leu Leu Lys Ser Val Leu Val Phe Ala Ala
Leu Ser Ser 1 5 10 15 Ala Ser Ser Leu Gln Ala Leu Pro Val Gly Asn
Pro Ala Glu Pro Ser 20 25 30 Leu Met Ile Asp Gly Ile Leu Trp Glu
Gly Phe Gly Gly Asp Pro Cys 35 40 45 Asp Pro Cys Thr Thr Trp Cys
Asp Ala Ile Ser Met Ile Met Gly Tyr 50 55 60 Tyr Gly Asp Phe Val
Phe Asp Arg Val Leu Lys Thr Asp Val Asn Lys 65 70 75 80 Glu Phe Gln
Met Gly Ala Lys Pro Thr Thr Thr Thr Gly Asn Ala Ala 85 90 95 Ala
Pro Ser Thr 100 47 100 PRT Chlamydia trachomatis 47 Met Lys Lys Leu
Leu Lys Ser Val Leu Val Phe Ala Ala Leu Ser Ser 1 5 10 15 Ala Ser
Ser Leu Gln Ala Leu Pro Val Gly Asn Pro Ala Glu Pro Ser 20 25 30
Leu Met Ile Asp Gly Ile Leu Trp Glu Gly Phe Gly Gly Asp Pro Cys 35
40 45 Asp Pro Cys Thr Thr Trp Cys Asp Ala Ile Ser Met Arg Met Gly
Tyr 50 55 60 Tyr Gly Asp Phe Val Phe Asp Arg Val Leu Glu Thr Asp
Val Asn Lys 65 70 75 80 Glu Phe His Met Gly Ala Lys Pro Thr Thr Asp
Thr Gly Asn Ser Ala 85 90 95 Ala Pro Leu Thr 100 48 100 PRT
Chlamydia trachomatis 48 Met Lys Lys Leu Leu Lys Ser Val Leu Val
Phe Ala Ala Leu Ser Ser 1 5 10 15 Ala Ser Ser Leu Gln Ala Leu Pro
Val Gly Asn Pro Ala Glu Pro Ser 20 25 30 Leu Met Ile Asp Gly Ile
Leu Trp Glu Gly Phe Gly Gly Asp Pro Cys 35 40 45 Asp Pro Cys Thr
Thr Trp Cys Asp Ala Ile Ser Met Ile Met Gly Tyr 50 55 60 Tyr Gly
Asp Phe Val Phe Asp Arg Val Leu Lys Thr Asp Val Asn Lys 65 70 75 80
Glu Phe His Met Gly Asp Lys Pro Thr Ser Thr Thr Gly Asn Ala Thr 85
90 95 Ala Pro Thr Thr 100 49 100 PRT Chlamydia trachomatis 49 Met
Lys Lys Leu Leu Lys Ser Val Leu Val Phe Ala Ala Leu Ser Ser 1 5 10
15 Ala Ser Ser Leu Gln Ala Leu Pro Val Gly Asn Pro Ala Glu Pro Ser
20 25 30 Leu Met Ile Asp Gly Ile Leu Trp Glu Gly Phe Gly Gly Asp
Pro Cys 35 40 45 Asp Pro Cys Thr Thr Trp Cys Asp Ala Ile Ser Met
Ile Met Gly Tyr 50 55 60 Tyr Gly Asp Phe Val Phe Asp Arg Val Leu
Lys Thr Asp Val Asn Lys 65 70 75 80 Glu Phe His Met Gly Asp Lys Pro
Thr Ala Thr Thr Gly Asn Ala Ala 85 90 95 Ala Pro Ser Thr 100 50 101
PRT Chlamydia trachomatis 50 Met Lys Lys Leu Leu Lys Ser Val Leu
Val Phe Ala Ala Leu Ser Ser 1 5 10 15 Ala Ser Ser Leu Gln Ala Leu
Pro Val Gly Asn Pro Ala Glu Pro Ser 20 25 30 Leu Met Ile Asp Gly
Ile Leu Trp Glu Gly Phe Gly Gly Asp Pro Cys 35 40 45 Asp Pro Cys
Thr Thr Trp Cys Asp Ala Ile Ser Met Ile Met Gly Tyr 50 55 60 Tyr
Gly Asp Phe Val Phe Asp Arg Val Leu Lys Thr Asp Val Asn Lys 65 70
75 80 Glu Phe Lys Met Gly Glu Ala Leu Ala Gly Ser Thr Gly Asn Thr
Thr 85 90 95 Ser Thr Leu Ser Lys 100 51 103 PRT Chlamydia
trachomatis 51 Met Lys Lys Leu Leu Lys Ser Val Leu Val Phe Ala Ala
Leu Ser Ser 1 5 10 15 Ala Ser Ser Leu Gln Ala Leu Pro Val Gly Asn
Pro Ala Glu Pro Ser 20 25 30 Leu Met Ile Asp Gly Ile Leu Trp Glu
Gly Phe Gly Gly Asp Pro Cys 35 40 45 Asp Pro Cys Thr Thr Trp Cys
Asp Ala Ile Ser Met Val Met Gly Tyr 50 55 60 Tyr Gly Asp Phe Val
Phe Asp Arg Val Leu Lys Thr Asp Val Asn Lys 65 70 75 80 Glu Phe Gln
Met Gly Ala Ala Pro Thr Thr Ser Asp Val Ala Ala Gly 85 90 95 Leu
Gln Asn Asp Pro Thr Ile 100 52 102 PRT Chlamydia trachomatis 52 Met
Lys Lys Leu Leu Lys Ser Val Leu Val Phe Ala Ala Leu Ser Ser 1 5 10
15 Ala Ser Ser Leu Gln Ala Leu Pro Val Gly Asn Pro Ala Glu Pro Ser
20 25 30 Leu Met Ile Asp Gly Ile Leu Trp Glu Gly Phe Gly Gly Asp
Pro Cys 35 40 45 Asp Pro Cys Thr Thr Trp Cys Asp Ala Ile Ser Met
Arg Met Gly Tyr 50 55 60 Tyr Gly Asp Phe Val Phe Asp Arg Val Leu
Lys Thr Asp Val Asn Lys 65 70 75 80 Glu Phe Gln Met Gly Ala Ala Pro
Thr Thr Arg Asp Val Ala Gly Leu 85 90 95 Glu Lys Asp Pro Val Val
100 53 97 PRT Chlamydia trachomatis 53 Met Lys Lys Leu Leu Lys Ser
Val Leu Val Phe Ala Ala Leu Ser Ser 1 5 10 15 Ala Ser Ser Leu Gln
Ala Leu Pro Val Gly Asn Pro Ala Glu Pro Ser 20 25 30 Leu Met Ile
Asp Gly Ile Leu Trp Glu Gly Phe Gly Gly Asp Pro Cys 35 40 45 Ala
Pro Cys Thr Thr Trp Cys Asp Ala Ile Ser Met Val Met Gly Tyr 50 55
60 Tyr Gly Asp Phe Val Phe Asp Arg Val Leu Lys Thr Asp Val Asn Lys
65 70 75 80 Glu Phe Gln Met Gly Ala Ala Pro Thr Thr Asn Asp Ala Ala
Pro Lys 85 90 95 Thr 54 102 PRT Chlamydia trachomatis 54 Met Lys
Lys Leu Leu Lys Ser Val Leu Val Phe Ala Ala Leu Ser Ser 1 5 10 15
Ala Ser Ser Leu Gln Ala Leu Pro Val Gly Asn Pro Ala Glu Pro Ser 20
25 30 Leu Met Ile Asp Gly Ile Leu Trp Glu Gly Phe Gly Gly Asp Pro
Cys 35 40 45 Asp Pro Cys Thr Thr Trp Cys Asp Ala Ile Ser Met Val
Met Gly Tyr 50 55 60 Tyr Gly Asp Phe Val Phe Asp Arg Val Leu Lys
Thr Asp Val Asn Lys 65 70 75 80 Glu Phe Gln Met Gly Ala Glu Pro Thr
Thr Ser Asp Thr Ala Gly Leu 85 90 95 Ser Asn Asp Pro Thr Thr 100 55
100 PRT Chlamydia trachomatis 55 Met Lys Lys Leu Leu Lys Ser Val
Ala Val Phe Val Ala Gly Ser Ser 1 5 10 15 Ala Ser Ser Leu His Ala
Leu Pro Val Gly Asn Pro Ala Glu Pro Ser 20 25 30 Leu Met Ile Asp
Gly Ile Leu Trp Glu Gly Phe Gly Gly Asp Pro Cys 35 40 45 Asp Pro
Cys Thr Thr Trp Cys Asp Ala Ile Ser Met Arg Met Gly Leu 50 55 60
Tyr Leu Asp Phe Val Phe Asp Arg Val Leu Lys Thr Asp Val Asn Lys 65
70 75 80 Gln Phe Glu Met Gly Ala Ala Pro Thr Gly Asp Ala Asp Leu
Thr Thr 85 90 95 Ala Pro Thr Pro 100 56 99 PRT Chlamydia pneumoniae
56 Met Lys Lys Leu Leu Lys Ala Val Leu Ala Phe Ala Phe Ala Gly Ser
1 5 10 15 Val Gly Ser Leu Gln Ala Leu Pro Val Gly Asn Pro Ala Ser
Asp Ser 20 25 30 Leu Leu Ile Asp Gly Thr Ile Trp Glu Gly Ala Ala
Gly Asp Pro Cys 35 40 45 Asp Pro Ala Thr Thr Trp Cys Asp Ala Ile
Ser Leu Arg Ala Gly Phe 50 55 60 Tyr Gly Asp Phe Val Tyr Asp Ile
Val Leu Lys Val Asp Ala Pro Lys 65 70 75 80 Thr Phe Ser Met Gly Ala
Lys Pro Thr Thr Thr Gly Asn Gly Ser Ala 85 90 95 Ala Ala Asn 57 100
PRT Chlamydia trachomatis 57 Cys Thr Ala Arg Glu Asn Pro Ala Tyr
Gly Arg His Met Gln Asp Ala 1 5 10 15 Glu Met Phe Thr Asn Ala Ala
Tyr Met Ala Leu Ile Asn Trp Asp Arg 20 25 30 Phe Asp Val Phe Cys
Thr Leu Gly Ala Thr Ser Gly Tyr Leu Lys Gly 35 40 45 Asn Ser Ala
Ser Phe Asn Leu Val Gly Leu Phe Gly Asp Asn Glu Asn 50 55 60 His
Ala Thr Val Ser Asp Ser Lys Leu Val Pro Asn Met Ser Leu Asp 65 70
75 80 Gln Ser Val Val Glu Leu Tyr Thr Asp Thr Thr Phe Ala Trp Ser
Ala 85 90 95 Gly Ala Arg Ala 100 58 65 PRT Chlamydia trachomatis 58
Leu Thr Ala Arg Glu Asn Pro Ala Tyr Gly Arg His Met Gln Asp Ala 1 5
10 15 Glu Met Phe Thr Asn Cys Ala Tyr Met Ala Leu Ile Asn Trp Asp
Arg 20 25 30 Phe Asp Val Phe Cys Thr Leu Gly Ala Ser Ser Gly Tyr
Leu Lys Gly 35 40 45 Asn Ser Ala Ser Phe Asn Leu Val Gly Leu Phe
Gly Asn Asn Glu Asn 50 55 60 Gln 65 59 134 PRT Chlamydia
trachomatis 59 Cys Thr Ala Arg Glu Asn Pro Ala Tyr Gly Arg His Met
Gln Asp Ala 1 5 10 15 Glu Met Phe Thr Asn Cys Ala Tyr Met Ala Leu
Ile Asn Trp Asp Arg 20 25 30 Phe Asp Val Phe Cys Thr Leu Gly Ala
Thr Ser Gly Tyr Leu Lys Gly 35 40 45 Asn Ser Ala Ser Phe Asn Leu
Val Gly Leu Phe Gly Asp Asn Glu Asn 50 55 60 Gln Lys Thr Val Lys
Ala Glu Ser Val Pro Asn Met Ser Phe Asp Gln 65 70 75 80 Ser Val Val
Glu Leu Tyr Thr Asp Thr Thr Phe Ala Trp Ser Val Gly 85 90 95 Ala
Arg Ala Thr Lys Val Ser Asn Gly Thr Phe Val Pro Asn Met Ser 100 105
110 Leu Asp Gln Ser Val Val Glu Leu Tyr Thr Asp Thr Ala Phe Ala Trp
115 120 125 Ser Val Gly Ala Arg Ala 130 60 99 PRT Chlamydia
trachomatis 60 Leu Thr Ala Arg Glu Asn Pro Ala Tyr Gly Arg His Met
Gln Asp Ala 1 5 10 15 Glu Met Phe Thr Asn Cys Ala Tyr Met Ala Leu
Ile Asn Trp Asp Arg 20 25 30 Phe Asp Val Phe Cys Thr Leu Gly Ala
Ser Ser Gly Tyr Leu Lys Gly 35 40 45 Asn Ser Ala Ser Phe Asn Leu
Val Gly Leu Phe Gly Asp Asn Glu Asn 50 55 60 Gln Ser Thr Val Lys
Thr Asn Ser Val Pro Asn Met Ser Leu Asp Gln 65 70 75 80 Ser Val Val
Glu Leu Tyr Thr Asp Thr Ala Phe Ser Trp Ser Val Gly
85 90 95 Ala Arg Ala 61 99 PRT Chlamydia trachomatis 61 Cys Thr Ala
Arg Glu Asn Pro Ala Tyr Gly Arg His Met Gln Asp Ala 1 5 10 15 Glu
Met Phe Thr Asn Ala Ala Tyr Met Ala Leu Ile Asn Trp Asp Arg 20 25
30 Phe Asp Val Phe Cys Thr Leu Gly Ala Thr Ser Gly Tyr Leu Lys Gly
35 40 45 Asn Ser Ala Ser Phe Asn Leu Val Gly Leu Phe Gly Asp Asn
Glu Asn 50 55 60 Gln Ser Thr Val Lys Lys Asp Ala Val Pro Asn Met
Ser Phe Asp Gln 65 70 75 80 Ser Val Val Glu Leu Tyr Thr Asp Thr Thr
Phe Ala Trp Ser Val Gly 85 90 95 Ala Arg Ala 62 99 PRT Chlamydia
trachomatis 62 Leu Val Glu Arg Thr Asn Pro Ala Tyr Gly Lys His Met
Gln Asp Ala 1 5 10 15 Glu Met Phe Thr Asn Cys Ala Tyr Thr Ala Leu
Ile Asn Trp Asp Arg 20 25 30 Phe Asp Val Phe Cys Thr Leu Gly Ala
Thr Ser Gly Tyr Leu Lys Gly 35 40 45 Asn Ser Ala Ser Phe Asn Leu
Val Gly Leu Phe Gly Asp Gly Val Asn 50 55 60 Ala Thr Lys Pro Ala
Ala Asp Ser Ile Pro Asn Val Gln Leu Asn Gln 65 70 75 80 Ser Val Val
Glu Leu Tyr Thr Asp Thr Thr Phe Ala Trp Ser Val Gly 85 90 95 Ala
Arg Ala 63 100 PRT Chlamydia trachomatis 63 Asn Val Ala Arg Pro Asn
Pro Ala Tyr Gly Lys His Met Gln Asp Ala 1 5 10 15 Glu Met Phe Thr
Asn Ala Ala Tyr Met Ala Leu Ile Asn Trp Asp Arg 20 25 30 Phe Asp
Val Phe Cys Thr Leu Gly Ala Thr Thr Gly Tyr Leu Lys Gly 35 40 45
Asn Ser Ala Ser Phe Asn Leu Val Gly Leu Phe Gly Thr Lys Thr Gln 50
55 60 Ser Ser Ser Phe Asn Thr Ala Lys Leu Ile Pro Asn Thr Ala Leu
Asp 65 70 75 80 Gln Ser Val Val Glu Leu Tyr Ile Asn Thr Thr Phe Ala
Trp Ser Val 85 90 95 Gly Ala Arg Ala 100 64 100 PRT Chlamydia
trachomatis 64 Asn Val Ala Arg Pro Asn Pro Ala Tyr Gly Lys His Met
Gln Asp Ala 1 5 10 15 Glu Met Phe Thr Asn Ala Ala Tyr Met Ala Leu
Ile Asn Trp Asp Arg 20 25 30 Phe Asp Val Phe Cys Thr Leu Gly Ala
Thr Thr Gly Tyr Leu Lys Gly 35 40 45 Asn Ser Ala Ser Phe Asn Leu
Val Gly Leu Phe Gly Thr Lys Thr Gln 50 55 60 Ser Ser Gly Phe Asp
Thr Ala Asn Ile Val Pro Asn Thr Ala Leu Asn 65 70 75 80 Gln Ala Val
Val Glu Leu Tyr Thr Asp Thr Thr Phe Ala Trp Ser Val 85 90 95 Gly
Ala Arg Ala 100 65 100 PRT Chlamydia trachomatis 65 Asn Val Ala Arg
Pro Asn Pro Ala Tyr Gly Lys His Met Gln Asp Ala 1 5 10 15 Glu Met
Phe Thr Asn Ala Ala Tyr Met Ala Leu Ile Asn Trp Asp Arg 20 25 30
Phe Asp Val Phe Cys Thr Leu Gly Ala Thr Thr Gly Tyr Leu Lys Gly 35
40 45 Asn Ser Ala Ser Phe Asn Leu Val Gly Leu Phe Gly Thr Lys Thr
Lys 50 55 60 Ser Ser Asp Phe Asn Thr Ala Lys Leu Val Pro Asn Ile
Ala Leu Asn 65 70 75 80 Arg Ala Val Val Glu Leu Tyr Thr Asp Thr Thr
Phe Ala Trp Ser Val 85 90 95 Gly Ala Arg Ala 100 66 100 PRT
Chlamydia trachomatis 66 Asn Val Ala Arg Pro Asn Pro Ala Tyr Gly
Lys His Met Gln Asp Ala 1 5 10 15 Glu Met Phe Thr Asn Ala Ala Tyr
Met Ala Leu Ile Asn Trp Asp Arg 20 25 30 Phe Asp Val Phe Cys Thr
Leu Gly Ala Thr Thr Gly Tyr Leu Lys Gly 35 40 45 Asn Ser Ala Ser
Phe Asn Leu Val Gly Leu Phe Gly Thr Lys Thr Gln 50 55 60 Ser Thr
Asn Phe Asn Thr Ala Lys Leu Val Pro Asn Thr Ala Leu Asn 65 70 75 80
Gln Ala Val Val Glu Leu Tyr Thr Asp Thr Thr Phe Ala Trp Ser Val 85
90 95 Gly Ala Arg Ala 100 67 96 PRT Chlamydia trachomatis 67 Ala
Ser Arg Glu Asn Pro Ala Tyr Gly Lys His Met Gln Asp Ala Glu 1 5 10
15 Met Phe Thr Asn Ala Ala Tyr Met Ala Leu Ile Asn Trp Asp Arg Phe
20 25 30 Asp Val Phe Cys Thr Leu Gly Ala Thr Ser Gly Tyr Leu Lys
Gly Asn 35 40 45 Ser Ala Ala Phe Asn Leu Val Gly Leu Phe Gly Arg
Asp Glu Thr Ala 50 55 60 Val Ala Ala Asp Asp Ile Pro Asn Val Ser
Leu Ser Gln Ala Val Val 65 70 75 80 Glu Leu Tyr Thr Asp Thr Ala Phe
Ala Trp Ser Val Gly Ala Arg Ala 85 90 95 68 100 PRT Chlamydia
pneumoniae 68 Tyr Thr Thr Ala Val Asp Arg Pro Asn Pro Ala Tyr Asn
Lys His Leu 1 5 10 15 His Asp Ala Glu Trp Phe Thr Asn Ala Gly Ile
Phe Ala Leu Ile Asn 20 25 30 Trp Asp Arg Phe Asp Val Phe Cys Thr
Leu Gly Ala Ser Asn Gly Ile 35 40 45 Arg Lys Gly Asn Ser Thr Ala
Phe Asn Leu Val Gly Leu Phe Gly Val 50 55 60 Lys Gly Thr Thr Val
Asn Ala Asn Glu Leu Pro Asn Val Ser Leu Ser 65 70 75 80 Asn Gly Val
Val Glu Leu Tyr Thr Asp Thr Ser Phe Ser Trp Ser Val 85 90 95 Gly
Ala Arg Ala 100 69 100 PRT Chlamydia trachomatis 69 Ala Leu Trp Glu
Cys Gly Cys Ala Thr Leu Gly Ala Ser Phe Gln Tyr 1 5 10 15 Ala Gln
Ser Lys Pro Lys Val Glu Glu Leu Asn Val Leu Cys Asn Ala 20 25 30
Ala Glu Phe Thr Ile Asn Lys Pro Lys Gly Tyr Val Gly Gln Glu Phe 35
40 45 Pro Leu Asp Leu Lys Ala Gly Thr Asp Gly Val Thr Gly Thr Lys
Asp 50 55 60 Ala Ser Ile Asp Tyr His Glu Trp Gln Ala Ser Leu Ala
Leu Ser Tyr 65 70 75 80 Arg Leu Asn Met Phe Thr Pro Tyr Ile Gly Val
Lys Trp Ser Arg Ala 85 90 95 Ser Phe Asp Ala 100 70 100 PRT
Chlamydia trachomatis 70 Ala Leu Trp Glu Cys Gly Cys Ala Thr Leu
Gly Ala Ser Phe Gln Tyr 1 5 10 15 Ala Gln Ser Lys Pro Lys Val Glu
Glu Leu Asn Val Leu Cys Asn Ala 20 25 30 Ala Glu Phe Thr Ile Asn
Lys Pro Lys Gly Tyr Val Gly Lys Glu Leu 35 40 45 Pro Leu Asp Leu
Thr Ala Gly Thr Asp Ala Ala Thr Gly Thr Lys Asp 50 55 60 Ala Ser
Ile Asp Tyr His Glu Trp Gln Ala Ser Leu Ala Leu Ser Tyr 65 70 75 80
Arg Leu Asn Met Phe Thr Pro Tyr Ile Gly Val Lys Trp Ser Arg Ala 85
90 95 Ser Phe Asp Ala 100 71 100 PRT Chlamydia trachomatis 71 Ala
Leu Trp Glu Cys Gly Cys Ala Thr Leu Gly Ala Ser Phe Gln Tyr 1 5 10
15 Ala Gln Ser Lys Pro Lys Val Glu Glu Leu Asn Val Leu Cys Asn Ala
20 25 30 Ala Glu Phe Thr Ile Asn Lys Pro Lys Gly Tyr Val Gly Lys
Glu Phe 35 40 45 Pro Leu Asp Leu Thr Ala Gly Thr Asp Ala Ala Thr
Gly Thr Lys Asp 50 55 60 Ala Ser Ile Asp Tyr His Glu Trp Gln Ala
Ser Leu Ala Leu Ser Tyr 65 70 75 80 Arg Leu Asn Met Phe Thr Pro Tyr
Ile Gly Val Lys Trp Ser Arg Ala 85 90 95 Ser Phe Asp Ala 100 72 100
PRT Chlamydia trachomatis 72 Ala Leu Trp Glu Cys Gly Cys Ala Thr
Leu Gly Ala Ser Phe Gln Tyr 1 5 10 15 Ala Gln Ser Lys Pro Lys Val
Glu Glu Leu Asn Val Leu Cys Asn Ala 20 25 30 Ala Glu Phe Thr Ile
Asn Lys Pro Lys Gly Tyr Val Gly Gln Glu Phe 35 40 45 Pro Leu Ala
Leu Ile Ala Gly Thr Asp Ala Ala Thr Gly Thr Lys Asp 50 55 60 Ala
Ser Ile Asp Tyr His Glu Trp Gln Ala Ser Leu Ala Leu Ser Tyr 65 70
75 80 Arg Leu Asn Met Phe Thr Pro Tyr Ile Gly Val Lys Trp Ser Arg
Ala 85 90 95 Ser Phe Asp Ala 100 73 100 PRT Chlamydia trachomatis
73 Ala Leu Trp Glu Cys Gly Cys Ala Thr Leu Gly Ala Ser Phe Gln Tyr
1 5 10 15 Ala Gln Ser Lys Pro Lys Val Glu Glu Leu Asn Val Leu Cys
Asn Ala 20 25 30 Ala Glu Phe Thr Ile Asn Lys Pro Lys Gly Tyr Val
Gly Lys Glu Phe 35 40 45 Pro Leu Asp Leu Thr Ala Gly Thr Asp Ala
Ala Thr Gly Thr Lys Asp 50 55 60 Ala Ser Ile Asp Tyr His Glu Trp
Gln Ala Ser Leu Ala Leu Ser Tyr 65 70 75 80 Arg Leu Asn Met Phe Thr
Pro Tyr Ile Gly Val Lys Trp Ser Arg Ala 85 90 95 Ser Phe Asp Ala
100 74 100 PRT Chlamydia trachomatis 74 Ala Leu Trp Glu Cys Gly Cys
Ala Thr Leu Gly Ala Ser Phe Gln Tyr 1 5 10 15 Ala Gln Ser Lys Pro
Lys Ile Glu Glu Leu Asn Val Leu Cys Asn Ala 20 25 30 Ala Glu Phe
Thr Ile Asn Lys Pro Lys Gly Tyr Val Gly Lys Glu Phe 35 40 45 Pro
Leu Asp Leu Thr Ala Gly Thr Asp Ala Ala Thr Gly Thr Lys Asp 50 55
60 Ala Ser Ile Asp Tyr His Glu Trp Gln Ala Ser Leu Ser Leu Ser Tyr
65 70 75 80 Arg Leu Asn Met Phe Thr Pro Tyr Ile Gly Val Lys Trp Ser
Arg Ala 85 90 95 Ser Phe Asp Ser 100 75 100 PRT Chlamydia
trachomatis 75 Ala Leu Trp Glu Cys Gly Cys Ala Thr Leu Gly Ala Ser
Phe Gln Tyr 1 5 10 15 Ala Gln Ser Lys Pro Lys Val Glu Glu Leu Asn
Val Leu Cys Asn Ala 20 25 30 Ser Glu Phe Thr Ile Asn Lys Pro Lys
Gly Tyr Val Gly Ala Glu Phe 35 40 45 Pro Leu Asn Ile Thr Ala Gly
Thr Glu Ala Ala Thr Gly Thr Lys Asp 50 55 60 Ala Ser Ile Asp Tyr
His Glu Trp Gln Ala Ser Leu Ala Leu Ser Tyr 65 70 75 80 Arg Leu Asn
Met Phe Thr Pro Tyr Ile Gly Val Lys Trp Ser Arg Val 85 90 95 Ser
Phe Asp Ala 100 76 100 PRT Chlamydia trachomatis 76 Ala Leu Trp Glu
Cys Gly Cys Ala Thr Leu Gly Ala Ser Phe Gln Tyr 1 5 10 15 Ala Gln
Ser Lys Pro Lys Val Glu Glu Leu Asn Val Leu Cys Asn Ala 20 25 30
Ser Glu Phe Thr Ile Asn Lys Pro Lys Gly Tyr Val Gly Ala Glu Phe 35
40 45 Pro Leu Asp Ile Thr Ala Gly Thr Glu Ala Ala Thr Gly Thr Lys
Asp 50 55 60 Ala Ser Ile Asp Tyr His Glu Trp Gln Ala Ser Leu Ala
Leu Ser Tyr 65 70 75 80 Arg Leu Asn Met Phe Thr Pro Tyr Ile Gly Val
Lys Trp Ser Arg Val 85 90 95 Ser Phe Asp Ala 100 77 100 PRT
Chlamydia trachomatis 77 Ala Leu Trp Glu Cys Gly Cys Ala Thr Leu
Gly Ala Ser Phe Gln Tyr 1 5 10 15 Ala Gln Ser Lys Pro Lys Val Glu
Glu Leu Asn Val Leu Cys Asn Ala 20 25 30 Ser Glu Phe Thr Ile Asn
Lys Pro Lys Gly Tyr Val Gly Ala Glu Phe 35 40 45 Pro Leu Asp Ile
Thr Ala Gly Thr Glu Ala Ala Thr Gly Thr Lys Asp 50 55 60 Ala Ser
Ile Asp Tyr His Glu Trp Gln Ala Ser Leu Ala Leu Ser Tyr 65 70 75 80
Arg Leu Asn Met Phe Thr Pro Tyr Ile Gly Val Lys Trp Ser Arg Val 85
90 95 Ser Phe Asp Ala 100 78 100 PRT Chlamydia trachomatis 78 Ala
Leu Trp Glu Cys Gly Cys Ala Thr Leu Gly Ala Ser Phe Gln Tyr 1 5 10
15 Ala Gln Ser Lys Pro Lys Val Glu Glu Leu Asn Val Leu Cys Asp Ala
20 25 30 Ser Glu Phe Thr Ile Asn Lys Pro Lys Gly Tyr Val Gly Ala
Glu Phe 35 40 45 Pro Leu Asp Ile Thr Ala Gly Thr Glu Ala Ala Thr
Gly Thr Lys Asp 50 55 60 Ala Ser Ile Asp Tyr His Glu Trp Gln Ala
Ser Leu Ala Leu Ser Tyr 65 70 75 80 Arg Leu Asn Met Phe Thr Pro Tyr
Ile Gly Val Lys Trp Ser Arg Val 85 90 95 Ser Phe Asp Ala 100 79 100
PRT Chlamydia trachomatis 79 Ala Leu Trp Glu Cys Gly Cys Ala Thr
Leu Gly Ala Ser Phe Gln Tyr 1 5 10 15 Ala Gln Ser Lys Pro Lys Val
Glu Glu Leu Asn Val Leu Cys Asn Ala 20 25 30 Ala Glu Phe Thr Ile
Asn Lys Pro Lys Gly Tyr Val Gly Gln Glu Phe 35 40 45 Pro Leu Asn
Ile Lys Ala Gly Thr Val Ser Ala Thr Asp Thr Lys Asp 50 55 60 Ala
Ser Ile Asp Tyr His Glu Trp Gln Ala Ser Leu Ala Leu Ser Tyr 65 70
75 80 Arg Leu Asn Met Phe Thr Pro Tyr Ile Gly Val Lys Trp Ser Arg
Ala 85 90 95 Ser Phe Asp Ala 100 80 100 PRT Chlamydia pneumoniae 80
Gly Leu Trp Glu Cys Gly Cys Ala Thr Leu Gly Glu Ser Phe Gln Tyr 1 5
10 15 Ala Gln Ser Lys Pro Lys Val Glu Glu Leu Asn Val Ile Cys Asn
Val 20 25 30 Ser Gln Phe Ser Val Asn Lys Pro Lys Gly Tyr Lys Gly
Val Ala Phe 35 40 45 Pro Leu Pro Thr Asp Ala Gly Val Ala Thr Ala
Thr Gly Thr Lys Ser 50 55 60 Ala Thr Ile Asn Tyr His Glu Trp Gln
Val Gly Ala Ser Leu Ser Tyr 65 70 75 80 Arg Leu Asn Ser Leu Val Pro
Tyr Ile Gly Val Gln Trp Ser Arg Ala 85 90 95 Thr Phe Asp Ala 100 81
94 PRT Chlamydia trachomatis 81 Asp Thr Ile Arg Ile Ala Gln Pro Lys
Ser Ala Thr Thr Val Phe Asp 1 5 10 15 Val Thr Thr Leu Asn Pro Thr
Ile Ala Gly Ala Gly Asp Val Lys Ala 20 25 30 Ser Ala Glu Gly Gln
Leu Gly Asp Thr Met Gln Ile Val Ser Leu Gln 35 40 45 Leu Asn Lys
Met Lys Ser Arg Lys Ser Cys Gly Ile Ala Val Gly Thr 50 55 60 Thr
Ile Val Asp Ala Asp Lys Tyr Ala Val Thr Val Glu Thr Arg Leu 65 70
75 80 Ile Asp Glu Arg Ala Ala His Val Asn Ala Gln Phe Arg Phe 85 90
82 92 PRT Chlamydia trachomatis 82 Asp Thr Ile Arg Ile Ala Gln Pro
Lys Ser Ala Glu Thr Ile Phe Asp 1 5 10 15 Val Thr Thr Leu Asn Pro
Thr Ile Ala Gly Ala Gly Asp Val Lys Thr 20 25 30 Ser Ala Glu Gly
Gln Leu Gly Asp Thr Met Gln Ile Val Ser Leu Gln 35 40 45 Leu Asn
Met Lys Ser Arg Lys Cys Gly Ile Ala Val Gly Thr Thr Ile 50 55 60
Val Asp Ala Asp Lys Tyr Ala Ile Thr Val Glu Thr Arg Leu Ile Asp 65
70 75 80 Glu Arg Ala Ala His Val Asn Ala Gln Phe Arg Phe 85 90 83
94 PRT Chlamydia trachomatis 83 Asp Thr Ile Arg Ile Ala Gln Pro Lys
Ser Ala Thr Ala Ile Phe Asp 1 5 10 15 Thr Thr Thr Leu Asn Pro Thr
Ile Ala Gly Ala Gly Asp Val Lys Thr 20 25 30 Gly Thr Glu Gly Gln
Leu Gly Asp Thr Met Gln Ile Val Ser Leu Gln 35 40 45 Leu Asn Lys
Met Lys Ser Arg Lys Ser Cys Gly Ile Ala Val Gly Thr 50 55 60 Thr
Ile Val Asp Ala Asp Lys Tyr Ala Val Thr Val Glu Thr Arg Leu 65 70
75 80 Ile Asp Glu Arg Ala Ala His Val Asn Ala Gln Phe Arg Phe 85 90
84 94 PRT Chlamydia trachomatis 84 Asp Thr Ile Arg Ile Ala Gln Pro
Lys Ser Ala Thr Ala Ile Phe Asp 1 5 10 15 Thr Thr Thr Leu Asn Pro
Thr Ile Ala Gly Ala Gly Asp Val Lys Ala 20 25 30 Ser Ala Glu Gly
Gln Leu Gly Asp Thr Met Gln Ile Val Ser Leu Gln 35 40
45 Leu Asn Lys Met Lys Ser Arg Lys Ser Cys Gly Ile Ala Val Gly Thr
50 55 60 Thr Ile Val Asp Ala Asp Lys Tyr Ala Val Thr Val Glu Thr
Arg Leu 65 70 75 80 Ile Asp Glu Arg Ala Ala His Val Asn Ala Gln Phe
Arg Phe 85 90 85 94 PRT Chlamydia trachomatis 85 Asp Thr Ile Arg
Ile Ala Gln Pro Lys Leu Ala Thr Ala Ile Phe Asp 1 5 10 15 Thr Thr
Thr Leu Asn Pro Thr Ile Ala Gly Ala Gly Asp Glu Lys Ala 20 25 30
Asn Ala Glu Gly Gln Leu Gly Asp Thr Met Gln Ile Val Ser Leu Gln 35
40 45 Leu Asn Lys Met Lys Ser Arg Lys Ser Cys Gly Ile Ala Val Gly
Thr 50 55 60 Thr Ile Val Asp Ala Asp Lys Tyr Ala Val Thr Val Glu
Thr Arg Leu 65 70 75 80 Ile Asp Glu Arg Ala Ala His Val Asn Ala Gln
Phe Arg Phe 85 90 86 95 PRT Chlamydia trachomatis 86 Asp Thr Ile
Arg Ile Ala Gln Pro Arg Leu Val Thr Pro Val Val Asp 1 5 10 15 Ile
Thr Thr Leu Asn Pro Thr Ile Ala Gly Ala Cys Asp Ser Lys Ala 20 25
30 Gly Asn Thr Glu Gly Gln Ile Ser Asp Thr Met Gln Ile Val Ser Leu
35 40 45 Gln Leu Asn Lys Met Lys Ser Arg Lys Ser Cys Gly Ile Ala
Val Gly 50 55 60 Thr Thr Ile Val Asp Ala Asp Lys Tyr Ala Val Thr
Val Glu Thr Arg 65 70 75 80 Leu Ile Asp Glu Arg Ala Ala His Val Asn
Ala Gln Phe Arg Phe 85 90 95 87 95 PRT Chlamydia trachomatis 87 Asp
Thr Ile Arg Ile Ala Gln Pro Lys Leu Ala Glu Ala Ile Leu Asp 1 5 10
15 Val Thr Thr Leu Asn Arg Thr Thr Ala Gly Lys Gly Ser Val Val Ser
20 25 30 Ala Gly Thr Asp Asn Glu Leu Ala Asp Thr Met Gln Ile Val
Ser Leu 35 40 45 Gln Leu Asn Lys Met Lys Ser Arg Lys Ser Cys Gly
Ile Ala Val Gly 50 55 60 Thr Thr Ile Val Asp Ala Asp Lys Tyr Ala
Val Thr Val Glu Ala Arg 65 70 75 80 Leu Ile Asp Glu Arg Ala Ala His
Val Asn Ala Gln Phe Arg Phe 85 90 95 88 94 PRT Chlamydia
trachomatis 88 Asp Thr Ile Arg Ile Ala Gln Pro Lys Leu Ala Lys Pro
Val Leu Asp 1 5 10 15 Thr Thr Thr Leu Asn Pro Thr Ile Ala Gly Lys
Gly Thr Val Val Ser 20 25 30 Ser Ala Glu Asn Glu Leu Ala Asp Thr
Met Gln Ile Val Ser Leu Gln 35 40 45 Leu Asn Lys Met Lys Ser Arg
Lys Ser Cys Gly Ile Ala Val Gly Thr 50 55 60 Thr Ile Val Asp Ala
Asp Lys Tyr Ala Val Thr Val Glu Thr Arg Leu 65 70 75 80 Ile Asp Glu
Arg Ala Ala His Val Asn Ala Gln Phe Arg Phe 85 90 89 94 PRT
Chlamydia trachomatis 89 Asp Thr Ile Arg Ile Ala Gln Pro Lys Leu
Ala Glu Ala Ile Leu Asp 1 5 10 15 Val Thr Thr Leu Asn Pro Thr Ile
Ala Gly Lys Gly Thr Val Val Ala 20 25 30 Ser Gly Ser Asp Asn Asp
Leu Ala Asp Thr Met Gln Ile Val Ser Leu 35 40 45 Gln Leu Asn Lys
Met Lys Ser Arg Lys Ser Cys Gly Ile Ala Val Gly 50 55 60 Thr Thr
Ile Val Asp Ala Asp Lys Tyr Ala Val Thr Val Glu Thr Arg 65 70 75 80
Leu Ile Asp Glu Arg Ala Ala His Val Asn Ala Gln Phe Arg 85 90 90 95
PRT Chlamydia trachomatis 90 Asp Thr Ile Arg Ile Ala Gln Pro Lys
Leu Ala Glu Ala Val Leu Asp 1 5 10 15 Val Thr Thr Leu Asn Pro Thr
Ile Ala Gly Lys Gly Ser Val Val Ala 20 25 30 Ser Gly Ser Glu Asn
Glu Leu Ala Asp Thr Met Gln Ile Val Ser Leu 35 40 45 Gln Leu Asn
Lys Met Lys Ser Arg Lys Ser Cys Gly Ile Ala Val Gly 50 55 60 Thr
Thr Ile Val Asp Ala Asp Lys Tyr Ala Val Thr Val Glu Thr Arg 65 70
75 80 Leu Ile Asp Glu Arg Ala Ala His Val Asn Ala Gln Phe Arg Phe
85 90 95 91 91 PRT Chlamydia trachomatis 91 Asp Thr Ile Arg Ile Ala
Gln Pro Lys Leu Glu Thr Ser Ile Leu Lys 1 5 10 15 Met Thr Thr Trp
Asn Pro Thr Ile Ser Gly Ser Gly Ile Asp Val Asp 20 25 30 Thr Lys
Ile Thr Asp Thr Leu Gln Ile Val Ser Leu Gln Leu Asn Lys 35 40 45
Met Lys Ser Arg Lys Ser Cys Leu Ile Ala Ile Gly Thr Thr Ile Val 50
55 60 Asp Ala Asp Lys Tyr Ala Val Thr Val Glu Thr Arg Leu Ile Asp
Glu 65 70 75 80 Arg Ala Ala His Val Asn Ala Gln Phe Arg Phe 85 90
92 93 PRT Chlamydia pneumoniae 92 Asp Asn Ile Arg Ile Ala Gln Pro
Lys Leu Pro Thr Ala Val Leu Asn 1 5 10 15 Leu Thr Ala Trp Asn Pro
Ser Leu Leu Gly Asn Ala Thr Ala Leu Ser 20 25 30 Thr Thr Asp Ser
Phe Ser Asp Phe Met Gln Ile Val Ser Cys Gln Ile 35 40 45 Asn Lys
Phe Lys Ser Arg Lys Ala Cys Val Thr Ala Val Ala Thr Leu 50 55 60
Ile Val Asp Ala Asp Lys Trp Ser Leu Thr Ala Glu Ala Arg Leu Asn 65
70 75 80 Asp Glu Arg Ala Ala His Ser Gly Ala Gln Phe Arg Phe 85 90
93 17 PRT Chlamydia pneumoniae 93 Cys Thr Gly Ser Ala Ala Ala Asn
Tyr Thr Thr Ala Val Asp Arg Pro 1 5 10 15 Asn 94 18 PRT Chlamydia
trachomatis 94 Cys Thr Gly Asp Ala Asp Leu Thr Thr Ala Pro Thr Pro
Ala Ser Arg 1 5 10 15 Glu Asn 95 19 PRT Chlamydia trachomatis 95
Cys Thr Thr Ala Thr Gly Asn Ala Ala Ala Pro Ser Thr Cys Thr Ala 1 5
10 15 Arg Glu Asn 96 17 PRT Chlamydia psitacci 96 Cys Ala Ser Gly
Thr Ala Ser Asn Thr Thr Val Ala Ala Asp Arg Ser 1 5 10 15 Asn 97 15
PRT Chlamydia pneumoniae 97 Cys Phe Gly Val Lys Gly Thr Thr Val Asn
Ala Asn Glu Leu Pro 1 5 10 15 98 15 PRT Chlamydia trachomatis 98
Cys Phe Gly Arg Asp Glu Thr Ala Val Ala Ala Asp Asp Ile Pro 1 5 10
15 99 18 PRT Chlamydia trachomatis 99 Cys Phe Gly Asp Asn Glu Asn
His Ala Thr Val Ser Asp Ser Lys Leu 1 5 10 15 Val Pro 100 14 PRT
Chlamydia psitacci 100 Cys Ile Gly Leu Ala Gly Thr Asp Phe Ala Asn
Gln Arg Pro 1 5 10 101 13 PRT Chlamydia pneumoniae 101 Cys Gln Ile
Asn Lys Phe Lys Ser Arg Lys Ala Cys Gly 1 5 10 102 13 PRT Chlamydia
trachomatis 102 Cys Gln Ile Asn Lys Met Lys Ser Arg Phe Ala Cys Gly
1 5 10 103 13 PRT Chlamydia trachomatis 103 Cys Gln Leu Asn Lys Met
Lys Ser Arg Lys Ala Cys Gly 1 5 10 104 13 PRT Chlamydia psitacci
104 Cys Gln Ile Asn Lys Phe Lys Ser Arg Phe Ala Cys Gly 1 5 10 105
6 PRT Chlamydia pneumoniae 105 Arg Lys Lys Glu Arg Ser 1 5 106 10
PRT Chlamydia pneumoniae 106 Ser Thr Glu Cys Asn Ser Gln Ser Pro
Gln 1 5 10 107 13 PRT Ebola virus 107 Asn Pro Asn Leu His Tyr Trp
Thr Thr Gln Asp Glu Gly 1 5 10 108 22 PRT Ebola virus 108 Ser Gly
Gln Ser Pro Ala Arg Thr Ser Ser Asp Pro Gly Thr Asn Thr 1 5 10 15
Thr Thr Glu Asp His Lys 20 109 9 PRT Ebola virus 109 Thr Gly Gly
Arg Arg Thr Arg Arg Glu 1 5 110 10 PRT Ebola virus 110 Arg Asp Arg
Phe Lys Arg Thr Ser Phe Phe 1 5 10 111 12 PRT Ebola virus 111 Glu
Gln His His Arg Arg Thr Asp Asn Asp Ser Thr 1 5 10 112 11 PRT Ebola
virus 112 Glu Asn Thr Asn Thr Ser Lys Ser Thr Asp Phe 1 5 10 113 12
PRT Ebola virus 113 Tyr Thr Ser Gly Lys Arg Ser Asn Thr Thr Gly Lys
1 5 10 114 11 PRT Ebola virus 114 Thr Thr Thr Ser Pro Gln Asn His
Ser Glu Thr 1 5 10 115 11 PRT Ebola virus 115 Pro Asp Gln Gly Asp
Asn Asp Asn Trp Trp Thr 1 5 10 116 12 PRT Ebola virus 116 Thr Ile
Ser Thr Ser Pro Gln Ser Leu Thr Thr Lys 1 5 10 117 16 PRT Ebola
virus 117 Thr Glu Asp Pro Ser Ser Gly Tyr Tyr Ser Thr Thr Ile Arg
Tyr Gln 1 5 10 15 118 15 PRT Ebola virus 118 Thr His His Gln Asp
Thr Gly Glu Glu Ser Ala Ser Ser Gly Lys 1 5 10 15
* * * * *