NOTCH mutations leading to increased receptor signaling

Abstract

The present invention is based upon the identification of regions within the NOTCH-1 receptor that, when mutated, lead to increased receptor signaling. The mutations are associated with uncontrolled cellular growth and this growth may be arrested using agents that interfere with NOTCH-1 activity, such as inhibitors of gamma-secretase. Assays for the NOTCH-1 mutations may be used diagnostically or as part of a treatment regimen for cancer patients.

Description

CROSS REFERENCE TO RELATED APPLICATIONS

[0001] The present application claims priority to, and the benefit of, U.S. provisional applications 60/598,546, filed on Aug. 4, 2004 and 60/672,053, filed on Apr. 18, 2005. The contents of these prior applications are hereby incorporated by reference in their entirety.

FIELD OF THE INVENTION

[0003] The present invention is directed to human NOTCH receptors that have undergone mutations resulting in either increased responsiveness to biological stimuli or constitutive activation independent of the need for other stimuli. The mutations occur in specific, well-defined, regions of NOTCH receptors and are somatically acquired by cancer cells as part of the process of malignant transformation. The growth of these cancer cells can be slowed or stopped using agents that interfere with receptor signaling.

BACKGROUND OF THE INVENTION

[0004] NOTCH receptors 1-4 are transmembrane receptor proteins that signal through a pathway that relies on regulated proteolysis. Following ligand-binding, the receptor is sequentially: i) cleaved extracellularly by metalloproteases of the ADAM family (Brou, et al., Mol. Cell 5:207-216 (2000); Mumm, et al. Mol Cell 5:197-206 (2000)); ii) mono-ubiquitinated on a lysine residue lying just internal to the transmembrane domain (Gupta-Rossi, et al., J. Cell Biol. 166:73-83 (2004)); iii) endocytosed (Gupta-Rossi, et al., J. Cell Biol. 166:73-83 (2004)), and iv) proteolytically cleaved by a gamma-secretase enzyme (De Strooper, et al., Nature 398:518-522 (1999)). This final step in the activation process permits the intracellular portion of NOTCH receptors to translocate to the cell nucleus where it interacts with transcription factors to alter gene activity. NOTCH receptor signaling appears to play an important role in the differentiation and proliferation of cells and in controlling apoptosis, three processes that are important with respect to neoplastic transformation (see U.S. Pat. No. 6,703,221).

[0005] The NOTCH-1 gene was discovered through its involvement in a (7;9) chromosomal translocation found in fewer than 1% of T-cell acute lymphoblastic leukemias (T-ALLs) (Ellisen, et al., Cell 66:649-661 (1991)). NOTCH-1 is highly expressed in thymocytes (Ellisen, et al., Cell 66:649-661(1991)), where it induces common lymphoid progenitors to adopt a T cell fate (Radtke, et al., Immunity 10:547-558). Subsequently, it promotes the assembly of pre-T cell receptor complexes which play a critical role in driving a proliferative burst that accompanies maturation of CD4-/CD8-thymocytes to the CD4+/CD8+ developmental stage (Wolfer, et al., Immunity 16:869-879 (2002)).

[0006] In its resting state, mature NOTCH-1 is a heterodimeric receptor comprised of a ligand-binding extracellular subunit (NEC) and a non-covalently associated transmembrane subunit (NTM) (Rand, et al., Mol. Cell. Biol. 20:1825-1835; Logeat, et al., Proc. Natl. Acad. Sc.i USA 95:8108-8112 (1998)). NEC consists of a ligand-binding domain comprised of epidermal growth factor-like repeats, three iterated Lin12/NOTCH repeats, and a conserved 103 amino acid sequence (hereafter termed HD, for heterodimerization domain) that is sufficient for association with the extracellular portion of NTM.

[0007] Physiologic activation of NOTCH receptors occurs when a ligand of the Delta-Serrate-Lag2 (DSL) family binds to the NEC subunit and initiates a cascade of successive proteolytic cleavages in the NTM subunit. The final cleavage, which is catalyzed by .gamma.-secretase, a multiprotein complex containing presenilin-1 or -2, nicastrin, APH-1, and PEN-2 (Francis, et al., Dev. Cell 3:85-97 (2002); Kimberly, et al., Proc. Natl. Acad. Sci. USA 100:6382-6387 (2003)) releases the intracellular part of NTM (called intracellular NOTCH, or ICN) from the membrane, permitting it to translocate to the cell nucleus. There, it associates with the DNA-binding factor CSL and co-activators of the Mastermind family to form a short-lived transcriptional activation complex (Wallberg, et al., Mol. Cell Biol. 22:7812-7819 (2002); Fryer, et al., Genes Dev. 16:1397-1411 (2002); Nam, et al., J. Biol. Chem. 278:21232-21239 (2003)) Degradation and turnover of the complex is apparently regulated by F-box factors of the SEL-10 family (Oberg, et al., J. Biol. Chem. 276:35847-35853 (2001)).

[0008] The (7;9) translocation creates a NOTCH-T cell receptor .beta. fusion gene that encodes N-terminally-deleted, constitutively active NOTCH-1 polypeptides similar to the ICN (Ellisen, et al., Cell 66:649-661 (1991); Aster, et al., Cold Spring Harb. Symp. Quant. Biol. 59:125-136 (1994); Das, et al., J. Biol. Chem., epublished May 3, 2004)) and these truncated and constitutively active forms of NOTCH-1 induce T-ALL in mouse models (Aster, et al., Mol. Cell Biol. 20:7505-7515 (2000)). NOTCH-1 is also the site of frequent retroviral insertions that cooperate with the E2A-PBX1 and cMYC transgenes in multistep pathways leading to the development of murine T-ALL (Hoemann, et al., Mol. Cell. Biol. 20:3831-3842 (2000); Feldman, et al., Blood 96:1906-1913(2000)). Further, NOTCH inhibitors cause a G.sub.O/G.sub.I cell cycle arrest in cell lines derived from human and murine NOTCH-1-associated T-ALLs, indicating that NOTCH signaling is required for the sustained growth of these leukemias (Weng, et al., Mol. Cell Biol. 23:655-664 (2003)).

[0009] The (7;9) translocation has only been associated with a very small percentage of patients having T-ALL. The identification of additional mutations associated with NOTCH-1 would have important implications for the pathogenesis of this type of cancer. Although mutations in NOTCH-2, NOTCH-3, and NOTCH-4 have not been identified in human cancer, it is known that abnormal increases in function of these NOTCH receptors in other mammals can cause T-ALL (NOTCH-2 and -3, Bellavia, et al., Embo J. 19:3337-3348 (2000); Rohn, J. Virol. 70:8071-8080 (1996); Weng, et al., Mol. Cell. Biol. 23:655-664)) and breast cancer (NOTCH-4, Callahan and Rafat, J. Mammary Gland Biol Neoplasia 6:23-36 (2001)). Identification of novel mutations in human tumors should be useful diagnostically in helping to identify the presence of cancer and in identifying cancer cells that respond to inhibitors of NOTCH signaling, thereby making it possible to direct rational cancer treatment with NOTCH signaling pathway inhibitors.

SUMMARY OF THE INVENTION

[0010] The present invention is based upon the discovery that mutations in two specific regions of the NOTCH-1 receptor lead to increased receptor signaling and are associated with uncontrolled cellular growth. The first is a region within the heterodimerization domain that is characterized by the presence of many amino acids that are highly conserved between species. The amino acids (residues 1571-1622 in SEQ ID NO:1, see FIGS. 1 and 3 for sequence identification numbers) are encoded by exon 26 and are found within the NEC subunit (i.e. the extracellular portion) of the NOTCH-1 protein. In intact NOTCH-1, they are non-covalently bound to amino acids encoded by exon 27 (amino acids 1674-1730) which lie on the NTM subunit (i.e., the transmembrane portion of the receptor). It is believed that mutations, especially non-conservative mutations, in these "hotspots" of the HD domain, i.e., 1571-1622 of the NEC subunit and 1674-1730 of the NTM subunit, destabilize the NOTCH-1 heterodimer. This leads to dissociation of NEC and successive metalloprotease cleavage, ubiquination, endocytosis, and cleavage by gamma-secretase, permitting nuclear translocation of the intracellular portion of NOTCH-1 and activation of target genes.

[0011] A second region involved by mutations that lead to uncontrolled cellular growth is the C-terminal portion of NOTCH-1 (amino acid 2311 to 2556), which includes the PEST region (amino acid 2423 to amino acid 2556). These mutations appear to reduce the rate at which the receptor is degraded intracellularly after activation, i.e., after it has been released due to gamma-secretase cleavage. As a result, the transcription of NOTCH-1-responsive genes continues for an inappropriately long period of time, again, leading to increased signaling and, ultimately, uncontrolled growth.

[0012] It has been discovered that mutations in the HD domain and in the PEST domain are often found in leukemic cell lines and in biological samples obtained from patients with T cell acute lymphoblastic leukemia (T-ALL). Uncontrolled growth in these cells may be arrested using agents that interfere with NOTCH signaling, such as gamma-secretase inhibitors. Overall, the experiments performed have led to the conclusion that there is a group of cancer cells that depend upon abnormal NOTCH activity to maintain uncontrolled growth and that mutations in the HD domain and in the PEST domain of NOTCH-1 may be used diagnostically to identify cancer cells likely to respond to gamma-secretase inhibitors or other factors that diminish NOTCH activity.

[0013] In its first aspect, the invention is directed to a substantially purified mutant NOTCH-1 receptor which has the amino acid sequence of wild type NOTCH-1, shown herein as SEQ ID NO:1, except for mutations within the hotspots of the HD domain. These mutations lead to increased receptor signaling relative to unmutated NOTCH-1, due to increased susceptibility to cleavage by gamma-secretase. The specific hotspot regions of SEQ ID NO:1 in which the mutations occur are amino acids 1571-1622 and/or amino acids 1674-1730.

[0014] As used herein, the term "substantially purified" means that the receptor protein (or the polynucleotide encoding the protein) has been separated from the biological components associated with NOTCH-1 as found in nature. Typically, a substantially pure protein (or polynucleotide) will comprise at least 85% of a sample, with greater percentages being preferred. Many means are available for assessing the purity of protein or nucleic acid samples, including analysis by electrophoresis, chromatography and analytical centrifugation. The term "unmutated NOTCH-1" refers to NOTCH-1 receptors having the sequence of SEQ ID NO:1 and to polynucleotides that encode receptors having this sequence. The term also includes allelic variations in these sequences that are normally found in cells and which are not associated with abnormal NOTCH-1 signaling or with abnormal, uncontrolled cellular growth.

[0015] In order to determine whether a mutation leads to increased gamma-secretase signaling, the assays described herein in the Examples section may be used. The most preferred method involves the use of cells transfected with a reporter gene, e.g., a polynucleotide encoding luciferase, that is under the control of a regulatory element turned on in response to NOTCH-1 signaling. Activation of the reporter gene upon exposure of cells to gamma-secretase can be readily measured and reflects NOTCH activity (see, e.g., Hsieh, et al., Mol. Cell. Biol. 16:952-959 (1996)).

[0016] Comparison of HD amino acid sequences from a variety of species has resulted in the identification of certain specific amino acid positions that are highly conserved in hotspots. Mutations in these positions, particularly non-conservative mutations, are particularly likely to destabilize the interaction between the NEC and NTM subunits of NOTCH-1. The term "non-conservative mutation" refers to a mutation in which wild type amino acids are replaced with amino acids having different chemical characteristics. Examples of a non-conservative substitution include the substitution of a proline for other amino acids within a well-structured protein domain (i.e., the HD domain), the substitution of a basic amino acid for a non-basic amino acid, the substitution of an acidic amino acid for a non-acidic amino acid, the substitution of a polar amino acid for a non-polar amino acid, etc. The distinction between conservative and non-conservative mutations is well recognized by those of skill in the art. Conserved amino acids in the HD domain where mutations are associated with increased NOTCH-1 signaling are found at positions: 1571; 1573; 1575; 1576; 1578; 1579; 1583; 1586; 1588; 1590; 1593; 1594; 1595; 1598; 1600-1607; 1610-1618; 1674; 1675; 1677; 1679-1684; 1686; 1695; 1696; 1703; 1706; 1715; 1720; 1725; and 1728. All of these positions are identified with reference to SEQ ID NO:1.

[0017] The invention includes specific mutations that are associated with the uncontrolled growth of cancer cells. With respect to the HD domain, this occurs in cases where amino acids 1571-1618 are replaced with a sequence selected from the group consisting of SEQ ID NO:12-SEQ ID NO:22, SEQ ID NO:102-118 or SEQ ID NO:145. Other mutations associated with the growth of cancer cells include the replacement of amino acids 1571-1622 with a sequence selected from the group consisting of SEQ ID NO:23-SEQ ID NO:29 or SEQ ID NO:119-127. Similarly, the sequence from amino acid 1674 to amino acid 1730 may be replaced with a sequence selected from the group consisting of SEQ ID NO:70-SEQ ID NO:75 or SEQ ID NO:128-137.

[0018] The inventors have also found that mutations in SEQ ID NO:1 occurring in the C-terminal portion of NOTCH-1, i.e., from amino acid 2311 to amino acid 2556, lead to increased NOTCH-1 signaling in response to exposure of cells to gamma-secretase. All of these mutations cause the appearance of a premature stop codon or a shift in the translational reading frame, and thus lead to the loss of some or all of the sequences corresponding to the PEST region (amino acid 2423 to 2556). It is believed that these mutations lead to a reduced rate at which receptor is degraded. Specific mutations include the replacement of the portion of SEQ ID NO:1 corresponding to amino acids 2311-2556 with a sequence selected from the group consisting of SEQ ID NO:30-SEQ ID NO:59 or SEQ ID NO:139-144.

[0019] Mutations in the HD domain and the PEST domain may occur independently of one another or may be present in a single NOTCH-1 receptor. For example, a substantially purified mutant NOTCH-1 receptor, may comprise or consist of the amino acid sequence of SEQ ID NO:1, but wherein: [0020] a) the receptor has at least one of the following mutations: [0021] i) the portion of said amino acid sequence of SEQ ID NO:1 from amino acid 1571 to amino acid 1618 is replaced with a sequence selected from the group consisting of SEQ ID NO:12-SEQ ID NO:22, SEQ ID NO:102-118, or SEQ ID NO:145; [0022] ii) the portion of the amino acid sequence of SEQ ID NO:1 from amino acid 1571 to amino acid 1622 is replaced with a sequence selected from the group consisting of SEQ ID NO:23-SEQ ID NO:29, SEQ ID NO:119-127; [0023] iii) the portion of the amino acid sequence of SEQ ID NO:1from amino acid 1674 to amino acid 1730 is replaced with a sequence selected from the group consisting of SEQ ID NO:70-SEQ ID NO:75 or SEQ ID NO:128-137; and [0024] b) and, in addition, the receptor is further mutated such that the portion of the amino acid sequence of SEQ ID NO:1 from amino acid 2311 to amino acid 2556 is replaced with a sequence selected from the group consisting of SEQ ID NO:30-SEQ ID NO:59 or SEQ ID NO:138-144.

[0025] Alternatively, a substantially purified mutant NOTCH-1 receptor, may comprise or consist of the amino acid sequence of SEQ ID NO:1, but wherein: [0026] a) the amino acid sequence of SEQ ID NO:1 is mutated in one or more amino acid positions selected from the group consisting of: 1571; 1573; 1575; 1576; 1578; 1579; 1583; 1586; 1588; 1590; 1593; 1594; 1595; 1598; 1600-1607; 1610-1618; 1674; 1675; 1677; 1679-1684; 1686; 1695; 1696; 1703; 1706; 1715; 1720; 1725; and 1728; and [0027] b) the amino acid sequence of SEQ ID NO:1 is further mutated such that the portion of said amino acid sequence from amino acid 2311 to amino acid 2556 is replaced with a sequence selected from the group consisting of SEQ ID NO:30-SEQ ID NO:59. When mutations are present in both the HD and PEST regions, this leads to receptors in which signaling in response to gamma-secretase is especially enhanced.

[0028] In another aspect, the invention includes polynucleotides that encode any of the mutant NOTCH-1 receptors described above. Nucleic acid sequences coding for the mutant receptors may be incorporated into a vector in which they are operably linked to a promoter. The term "operably linked" refers to genetic elements that are joined in a manner that enables them to carry out their normal functions. For example, a gene is operably linked to a promoter when its transcription is under the control of the promoter and the protein normally encoded by the gene is ultimately produced. The invention includes the vectors coding for the mutant receptors as well as host cells transformed with the vectors.

[0029] In another aspect, the invention is directed to methods of diagnostically testing a biological sample for the presence of cancer cells by assaying the sample for any of the mutant NOTCH-1 receptors described above. The term biological sample, as used herein includes all of the different types of samples commonly used in the art of medicine in making diagnoses. Thus, the term encompasses, biopsy samples of tissue or fluid, blood, plasma, serum, lymph, urine, etc. Depending on the type of cancer being considered, certain types of samples will be the ones predominantly used. For example, for leukemia, samples of blood, plasma, or serum would tend to be favored. The choice of appropriate samples for testing is routine in clinical medicine

[0030] Also included are the same methods applied to the NOTCH-2, NOTCH-3 and NOTCH-4 receptors, which contain regions with high levels of identity to the regions with mutations described for NOTCH-1. The NOTCH-1, -2, -3 and -4 receptors are very highly related and likely perform very similar or identical functions in a variety of cell types, differing mainly in that different members of this receptor family are expressed and used preferentially by different tissues and cell types. Cancers arising in different tissues may thus acquire very similar types of mutations that affect any of the NOTCH-1, -2, -3 or -4 receptors with identical consequences in terms of promoting tumor cell growth, depending on which of the receptor subtypes are expressed by the tissues from which these cancers arise. Thus, in this application, unless otherwise indicated, the term "NOTCH" receptor will be used to refer to diagnostic applications applied collectively to each of the NOTCH-1, -2, -3, and -4 receptors.

[0031] The presence of mutant receptor may be detected either by assays directed at the protein or by assays directed at the gene encoding the protein. With respect to assays directed at genes, the preferred method of testing is to amplify either the full-length polynucleotide encoding a mutant NOTCH receptor, or a portion of this polynucleotide, using the polymerase chain reaction (PCR). Oligonucleotide primers that can be used for amplification are described in the Examples section, but one of skill in the art can readily identify other appropriate primers based upon sequence information included herewith and known in the art.

[0032] Alternatively, gene mutations may be detected by performing hybridizations using probes under conditions that permit hybrids to form with mutant, but not wild type, nucleic acid. Both the assays and the oligonucleotides used in the assays are encompassed by the invention. The oligonucleotides should be at least 15 bases in length and hybridization assays should be carried out under stringent conditions. As used herein, the term "stringent conditions" refers to hybridizations carried out at approximately 60-70.degree. C. and at a low salt concentration (equivalent to, for example, 0.02-0.15 M NaCl).

[0033] Assays designed to detect the mutant protein per se may be carried out using antibodies that bind to mutant NOTCH receptors with at least a hundredfold greater affinity than to unmutated NOTCH receptors. These antibodies may be either monoclonal or polyclonal in nature and can be made by injecting an antibody-producing animal with peptides derived from regions of NOTCH receptor containing mutations. Antibodies made in this manner and which preferentially bind to mutant receptor are also a part of the invention.

[0034] The invention is also directed to methods for slowing or stopping the growth of cells with the mutant receptors described above by exposing the cells to an effective amount of an agent, such as a gamma-secretase inhibitor, that interferes with NOTCH receptor signaling. Examples of appropriate inhibitors include III-31-C, N-[N-(3,5-5 difluorophenacetyl)-L-alanyl]S-phenylglycine t-butyl ester) (DAPT), compound E, D-helical peptide 294, isocoumarins, BOC-Lys(Cbz)Ile-Leu-epoxide, and (Z-LL).sub.2-ketone (see Kornilova, et al., J. Biol. Chem. 278:16479-16473 (2003)). The term "effective amount" indicates a sufficient concentration of inhibitor or other agent to significantly reduce NOTCH-1 activity, for example, a sufficient amount to reduce activity by at least 20%. The presence of an effective amount of an agent that inhibits Notch receptor activity should also be reflected clinically. For example, the rate growth of cancer cells should be slowed to a significant extent (e.g., by at least 20%) relative to the rate of growth occurring prior to treatment.

[0035] Although gamma-secretase inhibitors have been found to be effective as therapeutics, other agents that inhibit NOTCH activity may also be used. The mutations described herein should cause several effects, including: i) increased metalloprotease cleavage; ii) increased mono-ubiquitination (an enzymatic step carried out by an E3 ligase); iii) increased endocytosis, iv) increased gamma-secretase cleavage (as already discussed); v) increased nuclear translocation; and vi) increased formation of a nuclear NOTCH transcriptional activation complex. Drugs that act at any of these steps should stop the growth of cancer cells. One agent, an inhibitor of the nuclear complex, a dominant negative form of a protein called mastermind-like-1, has been previously described and inhibits all four NOTCH receptor subtypes (Weng, et al., Mol Cell Biol 23:655-664 (2003)).

[0036] Also encompassed by the invention are methods of assaying a test compound for its ability to inhibit abnormal cellular growth induced by the presence of a mutated NOTCH receptor. This involves incubating the test compound with cells that express any of the mutant NOTCH-1 receptors described above. Such cells may either be isolated from a biological sample or created by transfecting a cell with an expression vector encoding the mutant receptor. The incubation is carried out in the presence of gamma-secretase and the amount of NOTCH receptor activity that occurs is determined. Any method for measuring receptor activity may be used, but the luciferase reporter gene assay described in Hsieh, et al. (Mol. Cell. Biol. 16:952-959 (1996)) is preferred. The results obtained are then compared with those from assays carried out under essentially the same conditions but in the absence of the test compound. A reduction in activity caused by the test compound suggests that it is a NOTCH receptor inhibitor and may be useful in limiting the growth of cells expressing the receptor. This can be confirmed directly by comparing the growth of such cells in the presence and the absence of increasing concentrations of the test compound.

[0037] In addition to being of value in assays designed to evaluate the ability of a test compound to inhibit NOTCH receptor activity, polynucleotides encoding mutant NOTCH receptors may serve as a source for probes that can be used in the hybridization assays discussed above and as controls in diagnostic assays. The mutant receptor protein can be used to generate antibodies specific for mutated forms of NOTCH and can serve as a control in immunoassays designed to either quantitate the amount of NOTCH receptor present in a sample or to distinguish between normal and mutated forms of receptor. Both the mutant proteins and the polynucleotides encoding the mutant proteins should also be of interest to researchers studying biological processes such as differentiation and apoptosis.

[0038] As discussed above the invention includes isolated nucleic acid molecules encoding mutant Notch-1 receptors. Such nucleic acid molecules may comprise a nucleotide sequence selected from the group consisting of: [0039] a) a sequence of nucleotides that encodes a mutant Notch-1 receptor comprising any of the mutant amino acid sequences described herein and wherein, except for the codons encoding the mutant amino acid(s), the nucleotide sequence is that of SEQ ID NO:76; [0040] b) a sequence of nucleotides that encodes a mutant Notch-1 receptor having any of the amino acid sequences described herein; [0041] c) a sequence of nucleotides degenerate with the mutant Notch-1 encoding sequence of (a) or (b).

[0042] In another aspect, the invention includes a method of determining the oncogenic potential of a biological sample by assaying the sample for the presence any of one or more mutant Notch-1 receptors as described herein. The presence of mutant receptors indicates increased oncogenic potential relative to a sample in which such mutant NOTCH-1 receptors are not present.

[0043] In addition, the invention includes a method for following the progress of a therapeutic regime designed to alleviate a condition characterized by abnormal expression of a mutant NOTCH-1 receptor comprising: [0044] a) assaying a sample from a subject to determine the level of mutant NOTCH-1 receptor (or nucleic acid encoding a mutant Notch-1 receptor) at a first time point; [0045] b) assaying the level of the same receptor (or nucleic acid) at a second time point; and [0046] c) comparing the level at the second time point to the level determined at the first time point. Decreasing levels of mutant protein or nucleic acid is an indication that a therapy is working. Increasing levels are an indication that a therapy is not working.

[0047] The invention also encompasses a method for determining regression, progression or onset of a pathological disorder characterized by dysfunctional NOTCH-1 signal transduction. The method involves incubating a sample obtained from a patient having or suspected of having the disorder with a complementary nucleic acid hybridization probe having a sequence of nucleotides that is substantially homologous to that encoding one or more mutant NOTCH-1 receptors as described herein. The probe should generally be designed to interact with nucleic acid encoding the mutant form of the receptor and not the normal form as shown in SEQ ID NO:1. Binding between the probe and any complimentary MRNA that may be present in the sample is used to indicate the regression, progression or onset of the pathological disorder in said patient. Alternatively, assays may be performed using antibody (preferably monoclonal antibody) specific for a mutant form of the receptor. As the level of mutant receptor or nucleic acid encoding mutant receptor increases the likelihood that the disorder is present or progressing also increases. Reduced or absent levels of mutant receptor or nucleic acid is an indication of disorder regression or absence.

BRIEF DESCRIPTION OF THE DRAWINGS

[0048] FIG. 1-1 to 1-7: Aligned Full length amino acid sequences of human NOTCH receptors. The sequences shown are: NOTCH-1 (SEQ ID NO:1); NOTCH-2 (SEQ ID NO:2); NOTCH-3 (SEQ ID NO:3); and NOTCH-4 (SEQ ID NO:4).

[0049] FIG. 2A-2C. Identification of T-ALL cell lines that require NOTCH signals for growth. (2A) Effect of the .gamma.-secretase inhibitor compound E (GSI, 1 .mu.M) or vehicle alone (mock) on the cell cycle distribution of sensitive cell lines. Cells were treated for 4-8 days, fixed in cold 70% ethanol, stained with propidium iodide, and analyzed by flow cytometry. Live cells were gated by forward/side scatter criteria. (2B) Rescue of GSI-induced cell cycle arrest by MSCV-GFP-ICN1 retrovirus. (2C) Induction of cell cycle arrest by dominant negative MSCV-GFP-Mastermind-like-1 retrovirus. In (2B) and (2C), control cultures were transduced with control empty MSCV-GFP retrovirus or the indicated test virus. Two days post-transduction, cultures were split in two and treated with either compound E (GSI) or DMSO vehicle (mock) for 7-10 days. Cultures were then harvested and stained with the DNA dye DRAQ5, which permits flow cytometric measurement of DNA content in unsorted GFP- and GFP+ subpopulations. A minimum of 15,000 events was collected to create each DNA content histogram.

[0050] FIG. 3A-3G2. HD and PEST domain mutations in .gamma.-secretase inhibitor-sensitive T-ALL cell lines and in primary human T-ALL samples. (3A) sequence conservation of the exon 26 HD domain mutational "hotspot" region among vertebrate NOTCH-1 receptors. Positions of recurrent mutations are in bold and italics. (3B) missense mutations and deletions identified in cell lines (underlined) and primary T-ALLs. One mutation SEQ ID NO:145, was associated with acute myelogenous leukemia. Mutation sites are in bold and italics. (3C) in-frame insertions in the HD domain identified in primary T-ALLs. (3D) PEST domain mutations. The amino acid sequences of mutated NOTCH-1 alleles in cell lines (underlined) and primary T-ALLs are shown. Stop codons are indicated by asterisks. (3E) Schematic representation of PEST domain mutations. The circles (white, primary T-ALLs; black, cell lines) correspond to the positions at which the NOTCH-1 polypeptide sequence terminates. Key: NEC, NOTCH-1 extracellular domain; LNR, Lin/NOTCH repeats; HD, heterodimerization domain; NTM, NOTCH transmem-brane subunit; TM, transmembrane domain; ANK, ankyrin repeat domain; TAD, transcriptional activation domain. In (A-D), amino acid residues correspond to those of human pro-NOTCH-1. (3F) Western blot analysis of lysates from NOTCH-dependent T-ALL cell lines. Whole cell extracts were analyzed with a NOTCH-1 antibody raised against the intracellular transcriptional activation domain (Aster, et al., Mol. Cell. Biol. 20:7505-15). Positions of pro-NOTCH-1 and the normal NTM subunit are denoted. The lower panel is a longer exposure of the same blot that accentuates the presence of additional NOTCH-1 polypeptides of smaller size than the normal NTM polypeptide. Each lane was loaded with 25 .mu.g of total protein. (3G) Sequence conservation and mutations in exon 27, encoding the HD domain of the NOTCH-1 TMT subunit. This subunit of NOTCH is noncovalently bound to exon 26, the HD domain of the NEC subunit. Mutation sites are in bold and italics. For TALL 04.36 the i in the sequence indicates the site of insertion of the 14 amino acids shown immediately below.

[0051] FIG. 4A-4B. HD and PEST domain mutations activate NOTCH-1 signaling synergistically. (4A) Reporter gene assays. U2OS cells were transiently co-transfected in 24-well format with the indicated pcDNA3 plasmids, a NOTCH-responsive luciferase reporter gene, and an internal Renilla luciferase internal control plasmid, as described previously (Aster, et al., Mol. Cell. Biol. 20:7505-15). Twenty-five ng of pcDNA3 plasmid was used per well, except for experiments with pcDNA3-ICN1, in which 5 ng of plasmid per well were used. Normalized luciferase activities in whole cell lysates were determined in triplicate and expressed relative to the activity in lysates prepared from cells transfected with the empty vector control. (4B) Model for synergistic NOTCH-1 activation by dual mutations involving both the HD and PEST domains.

[0052] FIG. 5-1 to 5-11: Full length human NOTCH-1 gene sequence (SEQ ID NO:76) and encoded amino acid sequence (SEQ ID NO:1).

[0053] FIG. 6-1 to 6-3: Compilation of HD and PEST domain mutations.

DETAILED DESCRIPTION INVENTION

[0054] The present invention is concerned with the identification of mutations in the human NOTCH-1 receptor that lead to an increase in receptor signaling. The mutations are associated with cancer cells that respond positively to treatment with agents that interfere with NOTCH-1 activation, such as gamma-secretase inhibitors and are located within two specific regions of the NOTCH-1 protein, the HD domain and the PEST domain. It is expected that analogous mutations in corresponding regions of NOTCH-2, NOTCH-3 and NOTCH-4 receptor subtypes should also lead to increased signaling and abnormal growth.

[0055] Assays designed to detect the presence of NOTCH-1 mutations, e.g., assays based upon PCR amplification, may be used diagnostically to identify tissue samples with cells in which abnormal NOTCH-1 receptor signaling is occurring and which are, as a result, more likely to exhibit uncontrolled growth. Cancers characterized by this particular abnormality should be amenable to treatment with agents that reduce NOTCH-1 signaling, such as gamma-secretase inhibitors. Any of the inhibitors described herein or which have been described in the art (for example, in connection with potential use in the treatment of Alzheimer's disease) may be used in treatment regimens. The diagnostic assays and therapeutic methods may be applied to other types of cancer besides T-ALL. Such cancers include but are not limited to: carcinomas of the breast, prostate, lung, pancreas, ovary, colon, bladder, kidney and GI tract, B cell leukemias, B and T cell lymphomas, Hodgkin's lymphoma, acute myelogenous leukemias, melanomas, neuroblastomas, medulloblastomas, glioblastoma multiforme and other glial cancers of the brain, and mesotheliomas.

I. Production of Polynucleotides Encoding Mutant NOTCH-1 Receptors

[0056] The full length amino acid sequence of human NOTCH-1 is shown in FIG. 1 and has been given the sequence identification number SEQ ID NO:1. This is the normnal, unmutated form of receptor. Mutations in the HD domain and PEST domain that have been associated with samples from cancer patients or cancer cell lines are shown in FIG. 3. It should be noted that certain positions in the hotspots of the HD domain are highly conserved among different species (see FIG. 3A) and are especially sensitive to mutations that lead to increased gamma-secretase-generated activity.

[0057] Many methods are available for producing polynucleotides that encode NOTCH-1 receptors having the mutated sequences shown herein (see, e.g., Sambrook, et al., Molecular Cloning: A Laboratory Manual, 2.sup.nd ed., Cold Spring Harbor Press (1989)). For example, a cDNA library prepared by reverse transcribing MRNA isolated from cells that express the mutant NOTCH gene may be screened using probes synthesized based upon the sequences shown in FIG. 1. Alternatively, the desired sequences may be obtained by PCR amplifying reverse transcribed RNA. Primers for PCR may be constructed using the sequences shown herein and specific oligonucleotides that have been found to be suitable for amplification are described in the Examples section. Confirmation that the correct sequence has been amplified and identification of the presence of specific mutations may be obtained by sequencing amplification products.

[0058] Alternatively, nucleic acids may be produced using standard chemical synthetic methods well known in the art. Once obtained, the polynucleotides can be used in recombinant methods for making receptor protein, expressed in cells for the purpose of screening for potential therapeutic agents, or used as controls in diagnostic assays.

II. Production of Mutant Receptor Protein

[0059] NOTCH-1 receptor protein can be synthesized using chemical methods or produced using the polynucleotides described above. In the latter case, the DNA sequence encoding the mutant receptor may be placed in a vector containing transcriptional and translational signals recognizable by an appropriate host. The cloned receptor sequences, in double-stranded form, are inserted into the expression vector in an operable linkage. Appropriate vectors and techniques for cloning and expressing receptor are well known in the art of molecular biology.

[0060] Expression vectors may be introduced into host cells, preferably mammalian host cells, by methods such as calcium phosphate precipitation, microinjection, electroporation or viral transfer. Cells expressing the recombinant mutant receptor can be selected using standard, well established methods. One simple method for confirming the presence of the mutant receptor nucleic acid in cells is to perform PCR amplification using primers known to flank the receptor-encoding nucleotide sequence. The presence of receptor may also be confirmed using functional assays of NOTCH-1 such as the luciferase assay described in the Examples section and in the art (see, Hsieh, et al., Mol. Cell. Biol. 16:952-959 (1996)).

[0061] Once cells producing recombinant mutant NOTCH-1 receptor have been identified, they may be used in assays designed to identify agents capable of reducing signaling, for example, by incubation with gamma-secretase, or in assays that measure the ability of NOTCH-1 inhibitors to arrest uncontrolled cellular growth. The NOTCH-1 receptor itself may be used in diagnostic assays or to generate antibodies specific for mutant receptor that can be used in such assays.

III. Antibodies to Mutant NOTCH Receptor

[0062] The present invention encompasses antibodies that bind specifically to mutant forms of NOTCH receptors and processes for producing such antibodies. Antibodies that "bind specifically to mutant NOTCH receptors" are defined as those that have at least a hundredfold greater affinity for the mutant form of the receptor than for the unmutated form. The process for producing such antibodies may involve either injecting the full length mutant receptor protein into an appropriate animal or, preferably, injecting short peptides that include regions where mutations occur. The peptides should be at least five amino acids in length and may be injected either individually or in combinations.

[0063] Methods for making and selecting antibodies are well known to those of skill in the art as evidenced by standard reference works such as Harlow, et al., Antibodies, A Laboratory Manual, Cold Spring Harbor Laboratory, N.Y. (1988); Klein, Immunology: The Science of Self-Nonself Discrimination (1982); Kennett, et al., Monoclonal Antibodies and Hybridomas: A New Dimension in Biological Analyses (1980); and Campbell, "Monoclonal Antibody Technology," in: Laboratory Techniques in Biochemistry and Molecular Biology (1984).

[0064] The term "antibody," as used herein, is meant to include intact molecules as well as fragments that retain their ability to bind antigen such as Fab and F(ab').sub.2 fragments. The term "antibody" is also defined as referring to both monoclonal antibodies and polyclonal antibodies. Polyclonal antibodies are derived from the sera of animals immunized with an appropriate antigen. Monoclonal antibodies can be prepared using hybridoma technology as taught by references such as: Hammerling, et al., in Monoclonal Antibodies and T-Cell Hybridomas, Elsevier, N.Y., pp. 563-681 (1981). In general, this technology involves immunizing an immunocompetent animal, typically a mouse, with either intact protein or a fragment derived therefrom. Splenocytes are then extracted from the immunized animal and are fused with suitable myeloma cells, such as SP.sub.2O cells. Thereafter, the resulting hybridoma cells are selectively maintained in HAT medium and then cloned by limiting dilution (Wands, et al., Gastroenterology 80:225-232 (1981)). The cells obtained through such selection may then be assayed to identify clones that secrete antibodies binding preferentially to mutant forms of NOTCH-1 or mutant forms of other NOTCH receptors.

[0065] The antibodies, or fragments of antibodies, of the present invention may be used to detect the presence of mutant NOTCH receptor proteins using a variety of immunoassays. For example, the antibodies may be used in radioimmunoassays or immunometric assays, also known as "two site" or "sandwich" assays (see Chard, "An Introduction to Radioimmune Assay and Related Techniques" in: Laboratory Techniques in Biochemistry and Molecular Biology, North Holland Publishing Co., N.Y. (1978)). In a typical immunometric assay, a quantity of unlabeled antibody is bound to a solid support that is insoluble in the fluid being tested, e.g., blood, lymph, cellular extracts, etc. After the initial binding of antigen to immobilized antibody, a quantity of detectably labeled second antibody (which may or may not be the same as the first) is added to permit detection and/or quantitation of bound antigen (see, e.g., Kirkham, et al., Radioimmune Assay Methods, pp. 199-206 (1970)). Many variations of these types of assays are known in the art and may be employed for the detection of mutant NOTCH receptors.

[0066] Antibodies to mutant NOTCH receptors may also be used in the purification of either the intact receptor or fragments of these receptors (see generally, Dean, et al., Affinity Chromatograph A Practical Approach, IRLP Press (1986)). Typically, antibody is immobilized on a chromatographic matrix such as Sepharose 4B. The matrix is then packed into a column and a preparation containing mutant receptor is passed through the column under conditions that promote binding, e.g., under conditions of low salt. The column is then washed and bound receptor is eluted using a buffer that promotes dissociation from antibody (e.g., a buffer having an altered pH or salt concentration). The eluted receptor protein may be transferred into a buffer of choice, e.g., by dialysis and either stored or used directly. Purified receptor may be used in the immunoassays described above or for the generation of antibodies for use in assays.

IV. Assay Methods

[0067] Diagnostic assays for determining whether a biological sample contains cancer cells in which the mutations described herein contribute to uncontrolled growth may be performed either using the immunoassays described above to analyze protein or, preferably, by assaying a sample for the presence of nucleic acid sequences encoding a mutant form of receptor. The preferred method for nucleic acid analysis is by PCR amplifying polynucleic acid that has been extracted from cells using methods well known in the art. Amplification 5 may be performed using the oligonucleotide primers described in the Examples section below or using other oligonucleotides based upon the sequence of NOTCH-1 shown as SEQ ID NO:1. The amplified NOTCH receptor product may then be sequenced to determine whether it carries a mutation leading to increased signaling.

[0068] Alternatively, hybridizations may be performed under stringent conditions using probes that bind only to mutant sequences. The probes should be a minimum of 15 nucleotides in length and must cover the portion of the polynucleotide sequence that encodes the mutated region of NOTCH-1. Stringent conditions would typically involve hybridization at a temperature of 60-70.degree. C. with a low salt concentration (e.g., 0.02 M to 0.15 M NaCl). Procedures for identifying and labeling probes as well as for carrying out hybridizations and analyzing results are well known in the art of molecular biology.

[0069] Host cells transformed with vectors encoding recombinant mutant NOTCH receptors may also be used in assays designed to identify test compounds that reduce inappropriate signaling by the receptor. The assays will typically involve incubating cells in the presence of gamma-secretase and test compound. NOTCH receptor signaling can then be quantitated using assays such as those described herein or using other assays that are known in the art (see also, Hsieh, et al., Mol. Cell. Biol. 16:952-959 (1996)). As a simple alternative, cells known to be expressing mutated forms of NOTCH-1 that lead to uncontrolled growth may be cultured in the presence and absence of gamma-secretase and test compound to determine the extent to which the compound is capable of arresting growth.

V. Treatment Methods

[0070] Gamma-secretase inhibitors have been developed by pharmaceutical companies interested in their potential use as a treatment for Alzheimer's disease. These same inhibitors can be used to treat cancer cells in cases in which uncontrolled growth is associated with the NOTCH receptor mutations described herein.

[0071] The therapeutic regimen involves first testing a biological sample containing cancer cells removed from a patient to determine whether a mutated form of a NOTCH receptor is present. Patients whose samples evidence the presence of mutations would then be treated using a gamma-secretase inhibitor or another agent that interferes with NOTCH receptor activity. The dosage administered will depend upon the particular condition being treated, the route of administration and clinical considerations that are well known in the art. Dosages can be gradually increased until a beneficial effect, e.g., a slowing of tumor growth, is detected. Agents may be provided in either single or multiple dosage regimens and may be given either alone or in conjunction with other therapeutic agents.

[0072] Treatment of mutant NOTCH receptor-associated cancers is compatible with any route of administration and dosage form. Depending upon the particular condition being treated, certain dosage forms will tend to be more convenient or effective than others. For example, topical administration may be preferred in treating skin cancers, whereas parenteral administration might be preferred for leukemias. Apart from parenteral and topical preparations, agents may be administered orally, perorally, internally, intranasally, rectally, vaginally, lingually and transdermally. Specific dosage forms include tablets, pills, capsules, powders, aerosols, suppositories, skin patches, parenterals and oral liquids including suspensions, solutions and emulsions. Sustained release dosage forms may also be used. All dosage forms may be prepared using methods that are standard in the art (see, e.g., Remington's Pharmaceutical Sciences, 16.sup.th ed., Easton, Pa. (1980)).

EXAMPLES

[0073] The present example provides results suggesting that approximately 65% of human T cell acute lymphoblastic leukemias (T-ALLs), including tumors from all major molecular oncogenic subtypes and those occurring in children and adults, have NOTCH-1 mutations.

[0074] Most of these mutations occur in regions encoding an extracellular heterodimerization domain and as truncations removing a C-terminal PEST destruction box. The mutations cause increased NOTCH-1 signaling, and T-ALL cell lines bearing such mutations are growth arrested by NOTCH-1 pathway inhibitors.

[0075] Introduction

[0076] T cell acute lymphoblastic leukemia (T-ALL) is an aggressive neoplasm that preferentially affects children and adolescents. It is commonly associated with acquired chromosomal translocations and other genetic or epigenetic abnormalities, which lead to aberrant expression of a select group of transcription factors, including the helix-loop-helix proteins TAL1, TAL2, LYL1, and BHLHB1, the orphan homeobox proteins HOX 11 and HOX11L2, and the fusion proteins MLL-ENL and CALM-AF10. Mis-expression of these proteins, alone or in combination, perturbs normal pathways of T cell development, growth, and survival, and defines subgroups of T-ALL with distinct gene expression profiles. Nevertheless, enforced expression of any one of these proteins alone in murine models is not sufficient to cause T-ALL, emphasizing the multistep molecular pathogenesis of this disease.

[0077] Materials and Methods

[0078] Cell Lines

[0079] All T-ALL cell lines were cultured in RPMI (Invitrogen) supplemented with 10% fetal bovine serum (Hyclone), 1 mM sodium pyruvate, 2 mM L-glutamine, 100 U/ml penicillin G, and 100 .mu./ml streptomycin at 37.degree. C. under 5% CO.sub.2. U2OS cells were grown in Dulbecco's modified Eagle's medium (DMEM) supplemented with 10% fetal bovine serum (Hyclone), 2 mM L-glutamine, 100 U/ml penicillin G, and 100 .mu./ml streptomycin at 37.degree. C. under 5% CO.sub.2.

[0080] Gamma-Secretase Inhibitor Treatment of Cell Lines

[0081] Gamma-secretase activity was inhibited by addition of compound E to growing cells to a final concentration of 1 .mu./M. Mock-treated cultures were exposed to vehicle only, DMSO at a final concentration of 0.01%. The following human T-ALL cell lines were tested for sensitivity to compound E: T-ALL-1, ALL-SIL, KOPT-K1, HPB-ALL, DND-41, CCRF-HSB2, RPMI8402, THP-6, BE-13, JM-Jurkat, Jurkat-E6, REX, CEM, LOUCY, DU-528, KE-37, SKW-3, MKB-1, MOLT-3, MOLT-4, MOLT-13, MOLT-15, MOLT-16, SUPT-7, SUPT-11, SUPT-13, P12-Ichikawa, KARPAS-45, DSMZ-PEER and PF-382. The murine cell line T-6E, which was derived from a T-ALL induced with a membrane-tethered form of truncated NOTCH-1 that requires gamma-secretase cleavage for activation, was used as a positive control.

[0082] Retroviruses

[0083] Construction of retroviral expression plasmids (MSCV-GFP-ICN1 and MSCV-dominant negative Mastermind-like-1-GFP), production of high-titer pseudotyped, amphotropic, replication-defective retroviruses and spinoculation procedure were performed as described previously (Weng, et al., Mol. Cell Biol. 23:655-664).

[0084] Cell Cycle Analysis

[0085] Cells were stained with either propidium iodide after overnight fixation in cold 70% ethanol, or DRAQ5 (Biostatus, Leicester, UK) in complete medium (as suggested by the manufacturer) in instances where GFP fluorescence and DNA content measurement were measured simultaneously. DNA content was ascertained by flow cytometry and cell cycle factors determined using multicycle software (Phoenix Flow Systems, San Diego).

[0086] NOTCH-1 Expression Plasmids

[0087] NOTCH-1 expression plasmids in pcDNA3 have been described previously. To create a generic PEST deletion, the insertion mutation in the PEST domain of the T-ALL cell line ALL-SIL was amplified by PCR, identified by sequencing, and then sub-cloned into pcDNA3-NOTCH-1. Site-directed mutagenesis of the HD domain was conducted using the QuikChange kit (Stratagene). Mutagenic PCR primers were designed using the Stratagene web tool.

[0088] Receptor Gene Assays

[0089] NOTCH-1 expression plasmids (pcDNA3) were transiently transfected with U2OS cells with (i) an artificial luciferase reporter gene under the control of a promoter containing CSL binding sites and (ii) an internal Renilla luciferase control gene, as described previously (Aster, et al., Mol. Cell Biol. 20:7505-7515 (2000)), using Lipofectarnine Plus (Invitrogen). Total DNA was kept constant by adding empty vector as needed. All transfections were carried out in triplicate, and each experiment was repeated at least three times. Cell lysates were harvested 44-48 hours post-transfection and luciferase assays were carried out using the Dual Luciferase Assay System (Promega) on a Turner Systems luminometer, as per the manufacturer's suggestions.

[0090] PCR Amplification of NOTCH-1 cDNAs

[0091] RNA isolated from T-ALL cell lines (1).mu./g was reverse transcribed into cDNA using oligo-dT priming and Superscript reverse transcriptase (Invitrogen) in 20 .mu.l reactions per the manufacturer's suggested conditions. PCR amplifications were performed using 1 .mu.l of cDNA as template, 20 pMol of 5'-TTTGAATTCGGGCTGGACTGTGCGGAGCA TGTACCCGA (SEQ ID NO:78) and 5'-TTTGGATCCTCCGGAATGCGGGCGATCTG GGACTGCA (SEQ ID NO:79) NOTCH-1-specific primers, 0.25 .mu.l of Ex-Taq Takara, Japan), 1.times. reaction buffer, 1.times.dNTPs and 7.5% DMSO in a 50 .mu.l reaction volume. Cycle parameters were: 94.degree. C. for three minutes; 30 cycles of 94.degree. C. for one minute; 60.degree. C. for thirty seconds; 72.degree. C. for two minutes; and 72.degree. C. for eight minutes. PCR products were purified on Qiagen columns, digested with Bam HI and Eco RI, ligated into pSP72 and used to transform DH5 alpha cells. Miniprep DNA from individual colonies was sequenced using T7 and SP6 primers.

[0092] Patient Material

[0093] Samples of cryopreserved lymphoblasts from 96 children and young adults with T-ALL treated at St. Jude Children's Research Hospital and Dana Farber Cancer Institute were obtained with informed consent at the time of diagnosis. Genomic DNA from each sample was extracted with a commercial kit (GENTRA) following the manufacturer's instructions. RNA was prepared from cryopreserved lymphoblasts with RNAaqueous reagents (Ambion) according to the manufacturer's instructions.

[0094] Quantitative RT-PCR

[0095] Quantitative RT-PCR analysis of HOX11, HOX11L2, TAL1, LYL1, TAL2 and BHLHB1 was performed as previously described (Ferrando, et al., Cancer Cell 1:75-87; Ferrando, et al., Lancet 363:535-536) using an ABI PRISM 7700 Sequence Detection System instrument (Perkin Elmer Applied Biosystems). RT-PCR-based detection of MLL-ENL and CALM-AF10 fusion transcripts was also performed as previously described (Ferrando, et al., Lancet 363:535-536; Asnafi, et al., Blood 102:1000-1006).

[0096] Genomic DNA Preparation

[0097] Genomic DNA was prepared from primary T-ALLs by extraction with a commercial kit (GENTRA) following the manufacturer's instructions. DNA was prepared from cell lines by SDS lysis/proteinase K digestion, followed by phenol:chloroform extraction and ethanol precipitation. DNAs were resuspended in TE buffer, spectrophotometrically quantified and stored at -20.degree. C.

[0098] Mutation Detection

[0099] Mutation detection in exons 26, 27, and 34 of NOTCH-1 was performed via PCR-based denaturing HPLC using a WAVE DNA fragment analysis system (Transgenomic) equipped with a DNASep HT cartridge. Amplicons for mutation analysis were prepared with the following primer combinations. The N-terminal region of the HD domain of NOTCH-1, encoded in exon 26 was divided into two amplicons. Amplicon HD-N1 was amplified by PCR using primers HD-NLFW1: 5'-AGCCCCCTGTACGACCAGTA (SEQ ID NO:80), HD-N1FW1: 5'-CTTGCGCAGCTCCTCCTC (SEQ ID NO:81), and by nested PCR using primers HD-N1FW2: 5'-GACCAGTACTGCAAGGACCA (SEQ ID NO:82), HD-N1RV2: 5'-TCCTCGCGGCCGTAGTAG (SEQ ID NO:83). Amplicon HD2 was amplified by PCR using primers HD-N2FW1: 5'-GTGCTGCACACCAACGTG (SEQ ID NO:84), HD-N2RV1: 5'-GAGGGCCCAGGAGAGTTG (SEQ ID NO:85), and by nested PCR using primers HD-N2FW2: 5'-GCACACGGCCAGCAGATGAT (SEQ ID NO:86), and HD-N2RV2: 5'-CGCCGGGTCTCACTCAC (SEQ ID NO:87).

[0100] The C terminal region of the HD domain of NOTCH 1, encoded in exon 27, was amplified by PCR using primers HD-CFW1: GTGGCGTCATGGGCCTCA (SEQ ID NO:88) and HD-CRV1: TAGCAACTGGCACAAACAGC (SEQ ID NO:89), and by nested PCR using primers HD-CFW2: CATGGGCCTCAGTGTCCT (SEQ ID NO:90) and HD-CRV2: GCACAAACAGCCAGCGTGTC (SEQ ID NO:91).

[0101] Sequences of exon 34 encoding the PEST domain and the continuous N-region containing the TAD of NOTCH-1 were divided into three amplicons. Amplicon PEST1 and PEST2 were amplified by PCR using primers PESTFW1: 5'-GCAGCATGGCATGGTAGG (SEQ ID NO:92), and PESTRV1: 5'-AACATGTGTTTTAAAAAGGCTCCTC (SEQ ID NO:93), and by nested PCR using primers PEST1FW: 5'-AAACATCCAGCAGCAGC AAA (SEQ ID NO:94), PEST1RV: 5'-CACAGGCGAGGAGTAGCTGTG (SEQ ID NO:95), and PEST2FW: 5'-GTGACCGCAGCCCAGTTC (SEQ ID NO:96), and PEST2RV: 5'-AAAGGAAGCCGGGGTCTC (SEQ ID NO:97). Amplicon TAD1 was amplified by PCR using primers TAD1FW1: 5'-AGACTGGCCCACCTCGTCTCT (SEQ ID NO:98), TAD1RV1: 5'-GCTCTCCACTCAGGAAGCTC (SEQ ID NO:99), and by nested PCR using primers TAD1FW2: 5'-CGTCTCTCCCACCTGCCTGT (SEQ ID NO:100), and TAD1RV2: 5'-CTGAGCTCACGCCAAGGT (SEQ ID NO:101). Before DHPLC analysis, nested PCR products were denatured at 95.degree. C. for five minutes and slowly ramped to 40.degree. C. using a Peltier Thermal Cycler (MJ Research, PTC-225) in order to allow the formation of homo- and hetero-duplexes in samples heterozygous for a mutant allele.

[0102] Fifteen to 30 microliters of annealed PCR fragments per sample were injected on to the DNASep HT cartridge for analysis. Products were eluted at a constant flow rate of 1.5 ml/min with a linear acetonitrile gradient determined by Navigator software (Transgenomic) based on the size and GC-content of the amplicon. The gradient was produced by combining 0.1 m trimethylammonium acetate (TEAA) buffer (pH 7) (Transgenomic) and Buffer B (0.1 m TEAA with 25% acetonitrile). The elution profiles of DNA fragments, monitored by the system's UV detector, were used to produce chromatographs. The analysis sequence for each sample was as follows: a 1.9-minute time lapse for the detector, a 0.1-minute loading stage with a 3% decrease in Buffer B, a 2.0-minute linear gradient with an increasing slope of 2% Buffer B per minute, a 0.1-minute cleaning stage using 75% acetonitrile, and a 0.1-minute equilibration before the next injection. Homo- and hetero-duplex peaks were detected between the initial injection peak, produced by residual nucleotides and primers in the reaction, and the wash stage.

[0103] Melting profiles for the nested PCR products, ranging from 276 to 452 based pairs, were constructed using the Navigator software. Appropriate partial denaturing temperatures for mutation detection, which produced percent helicities between 30% and 98% at each base contained in regions of interest, were predicted for all amplicons: PEST1, 66.5.degree. C.; PEST2, 65.0.degree. C.; HD1, 67.0.degree. C. and 64.5.degree. C.; HD2, 67.0.degree. C. and 64.0.degree. C.; and TADI, 64.5.degree. C. and 63.9.degree. C. Less stable heteroduplexes denature earlier than homoduplexes and, thus, appear first in elution profiles. Therefore, direct sequencing was performed on samples with chromatographs displaying species that eluted before normal homoduplexes.

[0104] Results

[0105] To evaluate the possible role of NOTCH-1 signaling in human T-ALL beyond the rare cases associated with the t(7;9), we first tested T-ALL cell lines lacking the t(7;9) for NOTCH dependency by treating these cells with a .gamma.-secretase inhibitor (Weng, et al., Mol. Cell. Biol. 23:655-64). We observed that five of thirty human T-ALL cell lines tested showed a G.sub.0/G.sub.1 cell cycle arrest, which equaled or exceeded that of the T6E cell line, a reference NOTCH-1-dependent murine T-ALL (FIG. 2A). The growth suppression caused by gamma-secretase inhibitor was abrogated by retroviral expression of ICN1 (FIG. 2B), which is the fragment released after .gamma.-secretase cleavage, and reproduced by retroviral expression of dominant negative Mastermind-like-1 (FIG. 2C). Taken together, these results indicated that the growth of these five cell lines depends on NOTCH-mediated signals.

[0106] Because NOTCH receptors can be activated by physical dissociation of the NOTCH extracellular domain (Rand et al., Mol. Cell. Biol. 20:1825-35; Kramer, Sci. STKE 2000, PE1), we reasoned that a newly identified HD domain of NOTCH-1 could be the site of gain-of-function mutations. One class of retroviral insertion observed in murine T-ALL causes deletion of the negative regulatory C-terminal PEST sequences (Hoemann, et al., Mol. Cell. Biol. 20:3831-42; Feldman, et al., Blood 96:1906-13), making this a second candidate region for oncogenic NOTCH-1 mutations. Remarkably, DNA sequencing of these genomic regions revealed both HD domain and PEST domain mutations in 4 of the 5 NOTCH-dependent cell lines. Missense mutations affecting the HD domain (encoded by exon 26 of NOTCH-1) involved residues that are invariant in vertebrate NOTCH-1 receptors, and caused non-conservative changes in amino acid residues (FIG. 3A-C). One cell line, DND-41, had two different HD domain mutations within the same NOTCH-1 allele.

[0107] The PEST mutations (found in exon 34) were short insertions or deletions causing shifts in reading frame that are predicted to result in partial or complete deletion of the negative regulatory, C-terminal PEST domain (FIGS. 3D and 3E). PCR amplification, cloning, and sequencing of cDNAs revealed that the HD domain and PEST domain mutations lay in cis in the same NOTCH-1 allele in each of the 4 cell lines tested. Normal NOTCH-1 cDNA clones were also identified in each cell line, indicating that both alleles are expressed. This is consistent with the results of Western blot analysis (FIG. 3F), which revealed that cell lines with exon 26 and 34 mutations contained a polypeptide of the expected size of NTM and additional aberrant polypeptides of slightly smaller size than NTM. In contrast, the cell line TALL-1, which lacks mutations in exons 26 and 34, contained only NOTCH-1 polypeptides of the expected sizes.

[0108] We extended our mutational analysis of the HD and PEST domains of NOTCH-1to cryopreserved primary T-ALL samples obtained from the bone marrow of ninety-six children and adolescents at the time of diagnosis. At least one mutation was identified in 49 tumors (51%); 20 tumors (20.8%) had HD domain mutations only, 15 tumors (15.6%) had PEST domain mutations only, and 14 tumors (14.6%) had mutations in both the HD and PEST domains. Mutations were seen in tumors associated with mis-expression of HOX11 (2 of 3 cases), HOX11L2 (10 of 13, or 77%), TAL1 (11 of 31, or 35%), LYL1 (7 of 14, or 50%), MLL-ENL (1 of 3), or CALM-AF10 (1 of 2), which together define the major molecular subtypes of T-ALL. The HD domain mutations in primary tumors were clustered in a "hotspot" spanning residues 1574-1622, and included each of the three L to P missense mutations originally identified in the NOTCH-dependent T-ALL cell lines, as well as deletions of 1-2 residues and short "in-frame" insertions (FIG. 3A-B). PEST domain mutations included insertions or deletions that induced a shift in reading frame, as well as point mutations that created premature stop codons (FIGS. 3D-E, and FIG. 6).

[0109] In contrast to T-ALLs, no mutations were observed in 89 primary B-ALL samples. Mutations were also absent from four remission bone marrow samples obtained from patients whose T-ALLs harbored NOTCH-1 mutations, indicating that these mutations are not present in the germline, but rather acquired within the malignant clones. Polymorphisms have not been detected within the HD domain region of NOTCH-1 (NCBI SNP database), leading us to further suspect that the mutations in conserved residues we detected in T-ALLs were likely to have functional consequences.

[0110] To address this issue directly, we studied the effects of common HD domain mutations, with and without a PEST domain deletion, on activation of a NOTCH-sensitive luciferase reporter gene in transient expression assays (FIG. 4A). Single L to P mutations within the HD domain at residues 1575, 1594, or 1601 caused a 3-9-fold stimulation of luciferase activity, while an isolated PEST deletion (corresponding to the mutation found in the ALL-SIL cell line that produces a truncation at amino acid 2471) resulted in an approximate 1.5-2-fold stimulation. More strikingly, each HD mutation and the same PEST domain truncation in cis resulted in 20-40-fold stimulations of transcriptional activation. In contrast, the same mutations in trans produced lower levels of stimulation that were close to the average of each mutation acting alone.

[0111] Additional reporter gene assays carried out as described in FIG. 4 have proven that all HD mutations tested to date convey increased function on NOTCH-1. The mutations tested in these functional assays are: TABLE-US-00001 V1577E Sequence ID NO: 102 L1586Q Sequence ID NO: 106 F1593S Sequence ID NO: 117 L1597H Sequence ID NO: 109 R1599P Sequence ID NO: 15 I1617N Sequence ID NO: 115 I1617T Sequence ID NO: 116 V1677D Sequence ID NO: 135 L1679P Sequence ID NO: 71 I1681N Sequence ID NO: 73 A1702P Sequence ID NO: 129 I1719T Sequence ID NO: 128 ins(RLGSLNIPYKIEAV) Sequence ID NO: 137

[0112] These data suggest that increased function will be a general feature of mutations found in the HD domain of NOTCH1.

[0113] The synergistic interaction of HD and PEST domain mutations in cis is consistent with a model in which (i) HD domain mutations enhance .gamma.-secretase cleavage and increase the rate of production of ICN1, and (ii) truncations removing the PEST domain of ICN1 increase the half-life of this activated form of NOTCH (FIG. 4B). The intermediate levels of activation produced by these mutations in trans presumably reflect competition between relatively weak and strong gain-of-function NOTCH-1 polypeptides for factors required for processing and signaling. The stimulatory effects of mutated transmembrane NOTCH-1 polypeptides were completely abrogated by a .gamma.-secretase inhibitor, indicating a requirement for proteolysis at the juxtamembrane site of mutated NTM subunits for signal transduction (FIG. 4A). In contrast, the stimulation produced by ICN1, which is constitutively nuclear, was unaffected by .gamma.-secretase inhibition (FIG. 4A).

[0114] More recently we have observed two mutations in tumors other than T-ALL. An insertional mutation in exon 26 of NOTCH-1 was found in a case of acute myelogenous leukemia that causes amino acid residues 1593-1594 to be changed from FL to LSL. This mutation stimulates the ability of NOTCH-1 to activate a CSL-sensitive luciferase gene in assays analogous to those shown in FIG. 4A, indicating that it increases NOTCH-1 function. A novel NOTCH-1 mutation in exon 27 was found in a case of Langerhans cell histiocytosis, in which a highly conserved residue, amino acid 1693, was changed from a C to an R. Together, these data support the prediction that mutations identical or functionally similar to those discovered in T-ALL will be found in other forms of human neoplasia.

[0115] Discussion

[0116] The prevalence of gain-of-function NOTCH-1 mutations, which are found in every major molecular oncogenic subtype of T-ALL, strongly supports a pre-eminent role for upregulated NOTCH signaling in the pathogenesis of this tumor. The existence of at least one cell line, TALL-1, that apparently requires NOTCH signals for growth, yet lacks NOTCH-1 HD domain and PEST region mutations, suggests that other mechanisms exist that account for pre-T cell transformation involving the NOTCH pathway. It is conceivable that activating mutations affecting other regions of NOTCH-1 may be identified in human T-ALL cases.

[0117] Prior work has shown that activation of NOTCH receptors is achieved primarily through release of the NEC subunit (Rand, et al., Mol. Cell. Biol. 20:1825-35; Kramer, Sci. STKE 2000, PE1), which normally represses proteolysis. Recombinant "mini-receptors" consisting of just the NEC HD domain and the extracellular portion of NTM form stable heterodimers after furin cleavage, indicating that critical inter-subunit contacts lie in these regions. Hence, HD domain mutations may act by destabilizing NEC/NTM heterodimers. It follows that similar effects might be imparted by mutations occurring in the analogous heterodimerization region of the NTM subunit.

[0118] Several factors may explain the high frequency of NOTCH-1 mutations in T-ALL. The absolute requirement for NOTCH-1 signals during several stages of normal early T cell development provides a functional basis for the frequent involvement of this pathway. Further, unlike the t(7;9), which is created by aberrant recombination during attempted V-D-J.beta. rearrangement in committed T cell progenitors, the more common point mutations and insertions described here could occur in multipotent hematopoietic progenitors, which also normally express NOTCH-1 (Calvi, et al., Nature 425,841-6). If so, such mutations would be predicted to induce daughter cells to adopt a T cell fate (Allman, et al., J. Exp. Med. 194:99-106) and thereby increase the pool of cells that are at risk for additional leukemogenic events, which could include additional mutations affecting the same NOTCH-1 allele, as well as the chromosomal, genetic and epigenetic events that lead to the mis-expression of other critical transcription factors. Analogous NOTCH mutations might be involved in the molecular pathogenesis of certain more common types of human cancer, as NOTCH receptors are expressed in many normal tissues and a variety of malignancies (Allenspach, et al., Cancer Biol. Ther. 1:466-76).

[0119] Our findings identify the NOTCH pathway as a rational target for molecular therapy in T-ALL. Although approximately 75% of patients are currently cured of this disease with very intensive and cytotoxic chemotherapy regimens (Pui, et al., N. Engl. J. Med. 350:1535-48), new therapies are needed for patients with refractory disease, and less toxic, more efficacious combinations of drugs would be beneficial to all T-ALL patients. The development of potent, specific inhibitors of .gamma.-secretase (Wolfe, Nat. Rev. Drug Discov. 1:859-66), due to the involvement of this protease in the pathogenesis of Alzheimer's disease, should expedite clinical trials of cancer therapies aimed at blocking NOTCH-1 activity.

[0120] All references cited herein are fully incorporated by reference. Having now fully described the invention, it will be understood by those of skill in the art that the invention may be practiced within a wide and equivalent range of conditions, parameters and the like, without affecting the spirit or scope of the invention or any embodiment thereof.

Sequence CWU 1

1

164 1 2556 PRT Homo sapiens 1 Met Pro Pro Leu Leu Ala Pro Leu Leu Cys Leu Ala Leu Leu Pro Ala 1 5 10 15 Leu Ala Ala Arg Gly Pro Arg Cys Ser Gln Pro Gly Glu Thr Cys Leu 20 25 30 Asn Gly Gly Lys Cys Glu Ala Ala Asn Gly Thr Glu Ala Cys Val Cys 35 40 45 Gly Gly Ala Phe Val Gly Pro Arg Cys Gln Asp Pro Asn Pro Cys Leu 50 55 60 Ser Thr Pro Cys Lys Asn Ala Gly Thr Cys His Val Val Asp Arg Arg 65 70 75 80 Gly Val Ala Asp Tyr Ala Cys Ser Cys Ala Leu Gly Phe Ser Gly Pro 85 90 95 Leu Cys Leu Thr Pro Leu Asp Asn Ala Cys Leu Thr Asn Pro Cys Arg 100 105 110 Asn Gly Gly Thr Cys Asp Leu Leu Thr Leu Thr Glu Tyr Lys Cys Arg 115 120 125 Cys Pro Pro Gly Trp Ser Gly Lys Ser Cys Gln Gln Ala Asp Pro Cys 130 135 140 Ala Ser Asn Pro Cys Ala Asn Gly Gly Gln Cys Leu Pro Phe Glu Ala 145 150 155 160 Ser Tyr Ile Cys His Cys Pro Pro Ser Phe His Gly Pro Thr Cys Arg 165 170 175 Gln Asp Val Asn Glu Cys Gly Gln Lys Pro Gly Leu Cys Arg His Gly 180 185 190 Gly Thr Cys His Asn Glu Val Gly Ser Tyr Arg Cys Val Cys Arg Ala 195 200 205 Thr His Thr Gly Pro Asn Cys Glu Arg Pro Tyr Val Pro Cys Ser Pro 210 215 220 Ser Pro Cys Gln Asn Gly Gly Thr Cys Arg Pro Thr Gly Asp Val Thr 225 230 235 240 His Glu Cys Ala Cys Leu Pro Gly Phe Thr Gly Gln Asn Cys Glu Glu 245 250 255 Asn Ile Asp Asp Cys Pro Gly Asn Asn Cys Lys Asn Gly Gly Ala Cys 260 265 270 Val Asp Gly Val Asn Thr Tyr Asn Cys Arg Cys Pro Pro Glu Trp Thr 275 280 285 Gly Gln Tyr Cys Thr Glu Asp Val Asp Glu Cys Gln Leu Met Pro Asn 290 295 300 Ala Cys Gln Asn Gly Gly Thr Cys His Asn Thr His Gly Gly Tyr Asn 305 310 315 320 Cys Val Cys Val Asn Gly Trp Thr Gly Glu Asp Cys Ser Glu Asn Ile 325 330 335 Asp Asp Cys Ala Ser Ala Ala Cys Phe His Gly Ala Thr Cys His Asp 340 345 350 Arg Val Ala Ser Phe Tyr Cys Glu Cys Pro His Gly Arg Thr Gly Leu 355 360 365 Leu Cys His Leu Asn Asp Ala Cys Ile Ser Asn Pro Cys Asn Glu Gly 370 375 380 Ser Asn Cys Asp Thr Asn Pro Val Asn Gly Lys Ala Ile Cys Thr Cys 385 390 395 400 Pro Ser Gly Tyr Thr Gly Pro Ala Cys Ser Gln Asp Val Asp Glu Cys 405 410 415 Ser Leu Gly Ala Asn Pro Cys Glu His Ala Gly Lys Cys Ile Asn Thr 420 425 430 Leu Gly Ser Phe Glu Cys Gln Cys Leu Gln Gly Tyr Thr Gly Pro Arg 435 440 445 Cys Glu Ile Asp Val Asn Glu Cys Val Ser Asn Pro Cys Gln Asn Asp 450 455 460 Ala Thr Cys Leu Asp Gln Ile Gly Glu Phe Gln Cys Ile Cys Met Pro 465 470 475 480 Gly Tyr Glu Gly Val His Cys Glu Val Asn Thr Asp Glu Cys Ala Ser 485 490 495 Ser Pro Cys Leu His Asn Gly Arg Cys Leu Asp Lys Ile Asn Glu Phe 500 505 510 Gln Cys Glu Cys Pro Thr Gly Phe Thr Gly His Leu Cys Gln Tyr Asp 515 520 525 Val Asp Glu Cys Ala Ser Thr Pro Cys Lys Asn Gly Ala Lys Cys Leu 530 535 540 Asp Gly Pro Asn Thr Tyr Thr Cys Val Cys Thr Glu Gly Tyr Thr Gly 545 550 555 560 Thr His Cys Glu Val Asp Ile Asp Glu Cys Asp Pro Asp Pro Cys His 565 570 575 Tyr Gly Ser Cys Lys Asp Gly Val Ala Thr Phe Thr Cys Leu Cys Arg 580 585 590 Pro Gly Tyr Thr Gly His His Cys Glu Thr Asn Ile Asn Glu Cys Ser 595 600 605 Ser Gln Pro Cys Arg His Gly Gly Thr Cys Gln Asp Arg Asp Asn Ala 610 615 620 Tyr Leu Cys Phe Cys Leu Lys Gly Thr Thr Gly Pro Asn Cys Glu Ile 625 630 635 640 Asn Leu Asp Asp Cys Ala Ser Ser Pro Cys Asp Ser Gly Thr Cys Leu 645 650 655 Asp Lys Ile Asp Gly Tyr Glu Cys Ala Cys Glu Pro Gly Tyr Thr Gly 660 665 670 Ser Met Cys Asn Ile Asn Ile Asp Glu Cys Ala Gly Asn Pro Cys His 675 680 685 Asn Gly Gly Thr Cys Glu Asp Gly Ile Asn Gly Phe Thr Cys Arg Cys 690 695 700 Pro Glu Gly Tyr His Asp Pro Thr Cys Leu Ser Glu Val Asn Glu Cys 705 710 715 720 Asn Ser Asn Pro Cys Val His Gly Ala Cys Arg Asp Ser Leu Asn Gly 725 730 735 Tyr Lys Cys Asp Cys Asp Pro Gly Trp Ser Gly Thr Asn Cys Asp Ile 740 745 750 Asn Asn Asn Glu Cys Glu Ser Asn Pro Cys Val Asn Gly Gly Thr Cys 755 760 765 Lys Asp Met Thr Ser Gly Tyr Val Cys Thr Cys Arg Glu Gly Phe Ser 770 775 780 Gly Pro Asn Cys Gln Thr Asn Ile Asn Glu Cys Ala Ser Asn Pro Cys 785 790 795 800 Leu Asn Gln Gly Thr Cys Ile Asp Asp Val Ala Gly Tyr Lys Cys Asn 805 810 815 Cys Leu Leu Pro Tyr Thr Gly Ala Thr Cys Glu Val Val Leu Ala Pro 820 825 830 Cys Ala Pro Ser Pro Cys Arg Asn Gly Gly Glu Cys Arg Gln Ser Glu 835 840 845 Asp Tyr Glu Ser Phe Ser Cys Val Cys Pro Thr Gly Trp Gln Ala Gly 850 855 860 Gln Thr Cys Glu Val Asp Ile Asn Glu Cys Val Leu Ser Pro Cys Arg 865 870 875 880 His Gly Ala Ser Cys Gln Asn Thr His Gly Gly Tyr Arg Cys His Cys 885 890 895 Gln Ala Gly Tyr Ser Gly Arg Asn Cys Glu Thr Asp Ile Asp Asp Cys 900 905 910 Arg Pro Asn Pro Cys His Asn Gly Gly Ser Cys Thr Asp Gly Ile Asn 915 920 925 Thr Ala Phe Cys Asp Cys Leu Pro Gly Phe Arg Gly Thr Phe Cys Glu 930 935 940 Glu Asp Ile Asn Glu Cys Ala Ser Asp Pro Cys Arg Asn Gly Ala Asn 945 950 955 960 Cys Thr Asp Cys Val Asp Ser Tyr Thr Cys Thr Cys Pro Ala Gly Phe 965 970 975 Ser Gly Ile His Cys Glu Asn Asn Thr Pro Asp Cys Thr Glu Ser Ser 980 985 990 Cys Phe Asn Gly Gly Thr Cys Val Asp Gly Ile Asn Ser Phe Thr Cys 995 1000 1005 Leu Cys Pro Pro Gly Phe Thr Gly Ser Tyr Cys Gln His Asp Val 1010 1015 1020 Asn Glu Cys Asp Ser Gln Pro Cys Leu His Gly Gly Thr Cys Gln 1025 1030 1035 Asp Gly Cys Gly Ser Tyr Arg Cys Thr Cys Pro Gln Gly Tyr Thr 1040 1045 1050 Gly Pro Asn Cys Gln Asn Leu Val His Trp Cys Asp Ser Ser Pro 1055 1060 1065 Cys Lys Asn Gly Gly Lys Cys Trp Gln Thr His Thr Gln Tyr Arg 1070 1075 1080 Cys Glu Cys Pro Ser Gly Trp Thr Gly Leu Tyr Cys Asp Val Pro 1085 1090 1095 Ser Val Ser Cys Glu Val Ala Ala Gln Arg Gln Gly Val Asp Val 1100 1105 1110 Ala Arg Leu Cys Gln His Gly Gly Leu Cys Val Asp Ala Gly Asn 1115 1120 1125 Thr His His Cys Arg Cys Gln Ala Gly Tyr Thr Gly Ser Tyr Cys 1130 1135 1140 Glu Asp Leu Val Asp Glu Cys Ser Pro Ser Pro Cys Gln Asn Gly 1145 1150 1155 Ala Thr Cys Thr Asp Tyr Leu Gly Gly Tyr Ser Cys Lys Cys Val 1160 1165 1170 Ala Gly Tyr His Gly Val Asn Cys Ser Glu Glu Ile Asp Glu Cys 1175 1180 1185 Leu Ser His Pro Cys Gln Asn Gly Gly Thr Cys Leu Asp Leu Pro 1190 1195 1200 Asn Thr Tyr Lys Cys Ser Cys Pro Arg Gly Thr Gln Gly Val His 1205 1210 1215 Cys Glu Ile Asn Val Asp Asp Cys Asn Pro Pro Val Asp Pro Val 1220 1225 1230 Ser Arg Ser Pro Lys Cys Phe Asn Asn Gly Thr Cys Val Asp Gln 1235 1240 1245 Val Gly Gly Tyr Ser Cys Thr Cys Pro Pro Gly Phe Val Gly Glu 1250 1255 1260 Arg Cys Glu Gly Asp Val Asn Glu Cys Leu Ser Asn Pro Cys Asp 1265 1270 1275 Ala Arg Gly Thr Gln Asn Cys Val Gln Arg Val Asn Asp Phe His 1280 1285 1290 Cys Glu Cys Arg Ala Gly His Thr Gly Arg Arg Cys Glu Ser Val 1295 1300 1305 Ile Asn Gly Cys Lys Gly Lys Pro Cys Lys Asn Gly Gly Thr Cys 1310 1315 1320 Ala Val Ala Ser Asn Thr Ala Arg Gly Phe Ile Cys Lys Cys Pro 1325 1330 1335 Ala Gly Phe Glu Gly Ala Thr Cys Glu Asn Asp Ala Arg Thr Cys 1340 1345 1350 Gly Ser Leu Arg Cys Leu Asn Gly Gly Thr Cys Ile Ser Gly Pro 1355 1360 1365 Arg Ser Pro Thr Cys Leu Cys Leu Gly Pro Phe Thr Gly Pro Glu 1370 1375 1380 Cys Gln Phe Pro Ala Ser Ser Pro Cys Leu Gly Gly Asn Pro Cys 1385 1390 1395 Tyr Asn Gln Gly Thr Cys Glu Pro Thr Ser Glu Ser Pro Phe Tyr 1400 1405 1410 Arg Cys Leu Cys Pro Ala Lys Phe Asn Gly Leu Leu Cys His Ile 1415 1420 1425 Leu Asp Tyr Ser Phe Gly Gly Gly Ala Gly Arg Asp Ile Pro Pro 1430 1435 1440 Pro Leu Ile Glu Glu Ala Cys Glu Leu Pro Glu Cys Gln Glu Asp 1445 1450 1455 Ala Gly Asn Lys Val Cys Ser Leu Gln Cys Asn Asn His Ala Cys 1460 1465 1470 Gly Trp Asp Gly Gly Asp Cys Ser Leu Asn Phe Asn Asp Pro Trp 1475 1480 1485 Lys Asn Cys Thr Gln Ser Leu Gln Cys Trp Lys Tyr Phe Ser Asp 1490 1495 1500 Gly His Cys Asp Ser Gln Cys Asn Ser Ala Gly Cys Leu Phe Asp 1505 1510 1515 Gly Phe Asp Cys Gln Arg Ala Glu Gly Gln Cys Asn Pro Leu Tyr 1520 1525 1530 Asp Gln Tyr Cys Lys Asp His Phe Ser Asp Gly His Cys Asp Gln 1535 1540 1545 Gly Cys Asn Ser Ala Glu Cys Glu Trp Asp Gly Leu Asp Cys Ala 1550 1555 1560 Glu His Val Pro Glu Arg Leu Ala Ala Gly Thr Leu Val Val Val 1565 1570 1575 Val Leu Met Pro Pro Glu Gln Leu Arg Asn Ser Ser Phe His Phe 1580 1585 1590 Leu Arg Glu Leu Ser Arg Val Leu His Thr Asn Val Val Phe Lys 1595 1600 1605 Arg Asp Ala His Gly Gln Gln Met Ile Phe Pro Tyr Tyr Gly Arg 1610 1615 1620 Glu Glu Glu Leu Arg Lys His Pro Ile Lys Arg Ala Ala Glu Gly 1625 1630 1635 Trp Ala Ala Pro Asp Ala Leu Leu Gly Gln Val Lys Ala Ser Leu 1640 1645 1650 Leu Pro Gly Gly Ser Glu Gly Gly Arg Arg Arg Arg Glu Leu Asp 1655 1660 1665 Pro Met Asp Val Arg Gly Ser Ile Val Tyr Leu Glu Ile Asp Asn 1670 1675 1680 Arg Gln Cys Val Gln Ala Ser Ser Gln Cys Phe Gln Ser Ala Thr 1685 1690 1695 Asp Val Ala Ala Phe Leu Gly Ala Leu Ala Ser Leu Gly Ser Leu 1700 1705 1710 Asn Ile Pro Tyr Lys Ile Glu Ala Val Gln Ser Glu Thr Val Glu 1715 1720 1725 Pro Pro Pro Pro Ala Gln Leu His Phe Met Tyr Val Ala Ala Ala 1730 1735 1740 Ala Phe Val Leu Leu Phe Phe Val Gly Cys Gly Val Leu Leu Ser 1745 1750 1755 Arg Lys Arg Arg Arg Gln His Gly Gln Leu Trp Phe Pro Glu Gly 1760 1765 1770 Phe Lys Val Ser Glu Ala Ser Lys Lys Lys Arg Arg Glu Pro Leu 1775 1780 1785 Gly Glu Asp Ser Val Gly Leu Lys Pro Leu Lys Asn Ala Ser Asp 1790 1795 1800 Gly Ala Leu Met Asp Asp Asn Gln Asn Glu Trp Gly Asp Glu Asp 1805 1810 1815 Leu Glu Thr Lys Lys Phe Arg Phe Glu Glu Pro Val Val Leu Pro 1820 1825 1830 Asp Leu Asp Asp Gln Thr Asp His Arg Gln Trp Thr Gln Gln His 1835 1840 1845 Leu Asp Ala Ala Asp Leu Arg Met Ser Ala Met Ala Pro Thr Pro 1850 1855 1860 Pro Gln Gly Glu Val Asp Ala Asp Cys Met Asp Val Asn Val Arg 1865 1870 1875 Gly Pro Asp Gly Phe Thr Pro Leu Met Ile Ala Ser Cys Ser Gly 1880 1885 1890 Gly Gly Leu Glu Thr Gly Asn Ser Glu Glu Glu Glu Asp Ala Pro 1895 1900 1905 Ala Val Ile Ser Asp Phe Ile Tyr Gln Gly Ala Ser Leu His Asn 1910 1915 1920 Gln Thr Asp Arg Thr Gly Glu Thr Ala Leu His Leu Ala Ala Arg 1925 1930 1935 Tyr Ser Arg Ser Asp Ala Ala Lys Arg Leu Leu Glu Ala Ser Ala 1940 1945 1950 Asp Ala Asn Ile Gln Asp Asn Met Gly Arg Thr Pro Leu His Ala 1955 1960 1965 Ala Val Ser Ala Asp Ala Gln Gly Val Phe Gln Ile Leu Ile Arg 1970 1975 1980 Asn Arg Ala Thr Asp Leu Asp Ala Arg Met His Asp Gly Thr Thr 1985 1990 1995 Pro Leu Ile Leu Ala Ala Arg Leu Ala Val Glu Gly Met Leu Glu 2000 2005 2010 Asp Leu Ile Asn Ser His Ala Asp Val Asn Ala Val Asp Asp Leu 2015 2020 2025 Gly Lys Ser Ala Leu His Trp Ala Ala Ala Val Asn Asn Val Asp 2030 2035 2040 Ala Ala Val Val Leu Leu Lys Asn Gly Ala Asn Lys Asp Met Gln 2045 2050 2055 Asn Asn Arg Glu Glu Thr Pro Leu Phe Leu Ala Ala Arg Glu Gly 2060 2065 2070 Ser Tyr Glu Thr Ala Lys Val Leu Leu Asp His Phe Ala Asn Arg 2075 2080 2085 Asp Ile Thr Asp His Met Asp Arg Leu Pro Arg Asp Ile Ala Gln 2090 2095 2100 Glu Arg Met His His Asp Ile Val Arg Leu Leu Asp Glu Tyr Asn 2105 2110 2115 Leu Val Arg Ser Pro Gln Leu His Gly Ala Pro Leu Gly Gly Thr 2120 2125 2130 Pro Thr Leu Ser Pro Pro Leu Cys Ser Pro Asn Gly Tyr Leu Gly 2135 2140 2145 Ser Leu Lys Pro Gly Val Gln Gly Lys Lys Val Arg Lys Pro Ser 2150 2155 2160 Ser Lys Gly Leu Ala Cys Gly Ser Lys Glu Ala Lys Asp Leu Lys 2165 2170 2175 Ala Arg Arg Lys Lys Ser Gln Asp Gly Lys Gly Cys Leu Leu Asp 2180 2185 2190 Ser Ser Gly Met Leu Ser Pro Val Asp Ser Leu Glu Ser Pro His 2195 2200 2205 Gly Tyr Leu Ser Asp Val Ala Ser Pro Pro Leu Leu Pro Ser Pro 2210 2215 2220 Phe Gln Gln Ser Pro Ser Val Pro Leu Asn His Leu Pro Gly Met 2225 2230 2235 Pro Asp Thr His Leu Gly Ile Gly His Leu Asn Val Ala Ala Lys 2240 2245 2250 Pro Glu Met Ala Ala Leu Gly Gly Gly Gly Arg Leu Ala Phe Glu 2255 2260 2265 Thr Gly Pro Pro Arg Leu Ser His Leu Pro Val Ala Ser Gly Thr 2270 2275 2280 Ser Thr Val Leu Gly Ser Ser Ser Gly Gly Ala Leu Asn Phe Thr 2285 2290 2295 Val Gly Gly Ser Thr Ser Leu Asn Gly Gln Cys Glu Trp Leu Ser 2300 2305 2310 Arg Leu Gln Ser Gly Met Val Pro Asn Gln Tyr Asn Pro Leu Arg 2315 2320 2325 Gly Ser Val Ala Pro Gly Pro Leu Ser Thr Gln Ala Pro Ser Leu 2330 2335 2340 Gln His Gly Met Val Gly Pro Leu His Ser Ser Leu Ala Ala Ser 2345 2350 2355 Ala Leu Ser Gln Met Met Ser Tyr Gln Gly Leu Pro Ser Thr Arg 2360 2365 2370 Leu Ala Thr Gln Pro His Leu Val Gln Thr Gln Gln Val Gln Pro 2375 2380 2385 Gln Asn Leu Gln Met Gln Gln Gln Asn Leu Gln Pro Ala Asn Ile 2390 2395 2400 Gln Gln Gln Gln Ser Leu Gln Pro Pro Pro Pro Pro Pro Gln Pro 2405

2410 2415 His Leu Gly Val Ser Ser Ala Ala Ser Gly His Leu Gly Arg Ser 2420 2425 2430 Phe Leu Ser Gly Glu Pro Ser Gln Ala Asp Val Gln Pro Leu Gly 2435 2440 2445 Pro Ser Ser Leu Ala Val His Thr Ile Leu Pro Gln Glu Ser Pro 2450 2455 2460 Ala Leu Pro Thr Ser Leu Pro Ser Ser Leu Val Pro Pro Val Thr 2465 2470 2475 Ala Ala Gln Phe Leu Thr Pro Pro Ser Gln His Ser Tyr Ser Ser 2480 2485 2490 Pro Val Asp Asn Thr Pro Ser His Gln Leu Gln Val Pro Glu His 2495 2500 2505 Pro Phe Leu Thr Pro Ser Pro Glu Ser Pro Asp Gln Trp Ser Ser 2510 2515 2520 Ser Ser Pro His Ser Asn Val Ser Asp Trp Ser Glu Gly Val Ser 2525 2530 2535 Ser Pro Pro Thr Ser Met Gln Ser Gln Ile Ala Arg Ile Pro Glu 2540 2545 2550 Ala Phe Lys 2555 2 2471 PRT Homo sapiens 2 Met Pro Ala Leu Arg Pro Ala Leu Leu Trp Ala Leu Leu Ala Leu Trp 1 5 10 15 Leu Cys Cys Ala Ala Pro Ala His Ala Leu Gln Cys Arg Asp Gly Tyr 20 25 30 Glu Pro Cys Val Asn Glu Gly Met Cys Val Thr Tyr His Asn Gly Thr 35 40 45 Gly Tyr Cys Lys Cys Pro Glu Gly Phe Leu Gly Glu Tyr Cys Gln His 50 55 60 Arg Asp Pro Cys Glu Lys Asn Arg Cys Gln Asn Gly Gly Thr Cys Val 65 70 75 80 Ala Gln Ala Met Leu Gly Lys Ala Thr Cys Arg Cys Ala Ser Gly Phe 85 90 95 Thr Gly Glu Asp Cys Gln Tyr Ser Thr Ser His Pro Cys Phe Val Ser 100 105 110 Arg Pro Cys Leu Asn Gly Gly Thr Cys His Met Leu Ser Arg Asp Thr 115 120 125 Tyr Glu Cys Thr Cys Gln Val Gly Phe Thr Gly Lys Glu Cys Gln Trp 130 135 140 Thr Asp Ala Cys Leu Ser His Pro Cys Ala Asn Gly Ser Thr Cys Thr 145 150 155 160 Thr Val Ala Asn Gln Phe Ser Cys Lys Cys Leu Thr Gly Phe Thr Gly 165 170 175 Gln Lys Cys Glu Thr Asp Val Asn Glu Cys Asp Ile Pro Gly His Cys 180 185 190 Gln His Gly Gly Thr Cys Leu Asn Leu Pro Gly Ser Tyr Gln Cys Gln 195 200 205 Cys Pro Gln Gly Phe Thr Gly Gln Tyr Cys Asp Ser Leu Tyr Val Pro 210 215 220 Cys Ala Pro Ser Pro Cys Val Asn Gly Gly Thr Cys Arg Gln Thr Gly 225 230 235 240 Asp Phe Thr Phe Glu Cys Asn Cys Leu Pro Gly Phe Glu Gly Ser Thr 245 250 255 Cys Glu Arg Asn Ile Asp Asp Cys Pro Asn His Arg Cys Gln Asn Gly 260 265 270 Gly Val Cys Val Asp Gly Val Asn Thr Tyr Asn Cys Arg Cys Pro Pro 275 280 285 Gln Trp Thr Gly Gln Phe Cys Thr Glu Asp Val Asp Glu Cys Leu Leu 290 295 300 Gln Pro Asn Ala Cys Gln Asn Gly Gly Thr Cys Ala Asn Arg Asn Gly 305 310 315 320 Gly Tyr Gly Cys Val Cys Val Asn Gly Trp Ser Gly Asp Asp Cys Ser 325 330 335 Glu Asn Ile Asp Asp Cys Ala Phe Ala Ser Cys Thr Pro Gly Ser Thr 340 345 350 Cys Ile Asp Arg Val Ala Ser Phe Ser Cys Met Cys Pro Glu Gly Lys 355 360 365 Ala Gly Leu Leu Cys His Leu Asp Asp Ala Cys Ile Ser Asn Pro Cys 370 375 380 His Lys Gly Ala Leu Cys Asp Thr Asn Pro Leu Asn Gly Gln Tyr Ile 385 390 395 400 Cys Thr Cys Pro Gln Gly Tyr Lys Gly Ala Asp Cys Thr Glu Asp Val 405 410 415 Asp Glu Cys Ala Met Ala Asn Ser Asn Pro Cys Glu His Ala Gly Lys 420 425 430 Cys Val Asn Thr Asp Gly Ala Phe His Cys Glu Cys Leu Lys Gly Tyr 435 440 445 Ala Gly Pro Arg Cys Glu Met Asp Ile Asn Glu Cys His Ser Asp Pro 450 455 460 Cys Gln Asn Asp Ala Thr Cys Leu Asp Lys Ile Gly Gly Phe Thr Cys 465 470 475 480 Leu Cys Met Pro Gly Phe Lys Gly Val His Cys Glu Leu Glu Ile Asn 485 490 495 Glu Cys Gln Ser Asn Pro Cys Val Asn Asn Gly Gln Cys Val Asp Lys 500 505 510 Val Asn Arg Phe Gln Cys Leu Cys Pro Pro Gly Phe Thr Gly Pro Val 515 520 525 Cys Gln Ile Asp Ile Asp Asp Cys Ser Ser Thr Pro Cys Leu Asn Gly 530 535 540 Ala Lys Cys Ile Asp His Pro Asn Gly Tyr Glu Cys Gln Cys Ala Thr 545 550 555 560 Gly Phe Thr Gly Val Leu Cys Glu Glu Asn Ile Asp Asn Cys Asp Pro 565 570 575 Asp Pro Cys His His Gly Gln Cys Gln Asp Gly Ile Asp Ser Tyr Thr 580 585 590 Cys Ile Cys Asn Pro Gly Tyr Met Gly Ala Ile Cys Ser Asp Gln Ile 595 600 605 Asp Glu Cys Tyr Ser Ser Pro Cys Leu Asn Asp Gly Arg Cys Ile Asp 610 615 620 Leu Val Asn Gly Tyr Gln Cys Asn Cys Gln Pro Gly Thr Ser Gly Val 625 630 635 640 Asn Cys Glu Ile Asn Phe Asp Asp Cys Ala Ser Asn Pro Cys Ile His 645 650 655 Gly Ile Cys Met Asp Gly Ile Asn Arg Tyr Ser Cys Val Cys Ser Pro 660 665 670 Gly Phe Thr Gly Gln Arg Cys Asn Ile Asp Ile Asp Glu Cys Ala Ser 675 680 685 Asn Pro Cys Arg Lys Gly Ala Thr Cys Ile Asn Gly Val Asn Gly Phe 690 695 700 Arg Cys Ile Cys Pro Glu Gly Pro His His Pro Ser Cys Tyr Ser Gln 705 710 715 720 Val Asn Glu Cys Leu Ser Asn Pro Cys Ile His Gly Asn Cys Thr Gly 725 730 735 Gly Leu Ser Gly Tyr Lys Cys Leu Cys Asp Ala Gly Trp Val Gly Ile 740 745 750 Asn Cys Glu Val Asp Lys Asn Glu Cys Leu Ser Asn Pro Cys Gln Asn 755 760 765 Gly Gly Thr Cys Asp Asn Leu Val Asn Gly Tyr Arg Cys Thr Cys Lys 770 775 780 Lys Gly Phe Lys Gly Tyr Asn Cys Gln Val Asn Ile Asp Glu Cys Ala 785 790 795 800 Ser Asn Pro Cys Leu Asn Gln Gly Thr Cys Phe Asp Asp Ile Ser Gly 805 810 815 Tyr Thr Cys His Cys Val Leu Pro Tyr Thr Gly Lys Asn Cys Gln Thr 820 825 830 Val Leu Ala Pro Cys Ser Pro Asn Pro Cys Glu Asn Ala Ala Val Cys 835 840 845 Lys Glu Ser Pro Asn Phe Glu Ser Tyr Thr Cys Leu Cys Ala Pro Gly 850 855 860 Trp Gln Gly Gln Arg Cys Thr Ile Asp Ile Asp Glu Cys Ile Ser Lys 865 870 875 880 Pro Cys Met Asn His Gly Leu Cys His Asn Thr Gln Gly Ser Tyr Met 885 890 895 Cys Glu Cys Pro Pro Gly Phe Ser Gly Met Asp Cys Glu Glu Asp Ile 900 905 910 Asp Asp Cys Leu Ala Asn Pro Cys Gln Asn Gly Gly Ser Cys Met Asp 915 920 925 Gly Val Asn Thr Phe Ser Cys Leu Cys Leu Pro Gly Phe Thr Gly Asp 930 935 940 Lys Cys Gln Thr Asp Met Asn Glu Cys Leu Ser Glu Pro Cys Lys Asn 945 950 955 960 Gly Gly Thr Cys Ser Asp Tyr Val Asn Ser Tyr Thr Cys Lys Cys Gln 965 970 975 Ala Gly Phe Asp Gly Val His Cys Glu Asn Asn Ile Asn Glu Cys Thr 980 985 990 Glu Ser Ser Cys Phe Asn Gly Gly Thr Cys Val Asp Gly Ile Asn Ser 995 1000 1005 Phe Ser Cys Leu Cys Pro Val Gly Phe Thr Gly Ser Phe Cys Leu 1010 1015 1020 His Glu Ile Asn Glu Cys Ser Ser His Pro Cys Leu Asn Glu Gly 1025 1030 1035 Thr Cys Val Asp Gly Leu Gly Thr Tyr Arg Cys Ser Cys Pro Leu 1040 1045 1050 Gly Tyr Thr Gly Lys Asn Cys Gln Thr Leu Val Asn Leu Cys Ser 1055 1060 1065 Arg Ser Pro Cys Lys Asn Lys Gly Thr Cys Val Gln Lys Lys Ala 1070 1075 1080 Glu Ser Gln Cys Leu Cys Pro Ser Gly Trp Ala Gly Ala Tyr Cys 1085 1090 1095 Asp Val Pro Asn Val Ser Cys Asp Ile Ala Ala Ser Arg Arg Gly 1100 1105 1110 Val Leu Val Glu His Leu Cys Gln His Ser Gly Val Cys Ile Asn 1115 1120 1125 Ala Gly Asn Thr His Tyr Cys Gln Cys Pro Leu Gly Tyr Thr Gly 1130 1135 1140 Ser Tyr Cys Glu Glu Gln Leu Asp Glu Cys Ala Ser Asn Pro Cys 1145 1150 1155 Gln His Gly Ala Thr Cys Ser Asp Phe Ile Gly Gly Tyr Arg Cys 1160 1165 1170 Glu Cys Val Pro Gly Tyr Gln Gly Val Asn Cys Glu Tyr Glu Val 1175 1180 1185 Asp Glu Cys Gln Asn Gln Pro Cys Gln Asn Gly Gly Thr Cys Ile 1190 1195 1200 Asp Leu Val Asn His Phe Lys Cys Ser Cys Pro Pro Gly Thr Arg 1205 1210 1215 Gly Leu Leu Cys Glu Glu Asn Ile Asp Asp Cys Ala Arg Gly Pro 1220 1225 1230 His Cys Leu Asn Gly Gly Gln Cys Met Asp Arg Ile Gly Gly Tyr 1235 1240 1245 Ser Cys Arg Cys Leu Pro Gly Phe Ala Gly Glu Arg Cys Glu Gly 1250 1255 1260 Asp Ile Asn Glu Cys Leu Ser Asn Pro Cys Ser Ser Glu Gly Ser 1265 1270 1275 Leu Asp Cys Ile Gln Leu Thr Asn Asp Tyr Leu Cys Val Cys Arg 1280 1285 1290 Ser Ala Phe Thr Gly Arg His Cys Glu Thr Phe Val Asp Val Cys 1295 1300 1305 Pro Gln Met Pro Cys Leu Asn Gly Gly Thr Cys Ala Val Ala Ser 1310 1315 1320 Asn Met Pro Asp Gly Phe Ile Cys Arg Cys Pro Pro Gly Phe Ser 1325 1330 1335 Gly Ala Arg Cys Gln Ser Ser Cys Gly Gln Val Lys Cys Arg Lys 1340 1345 1350 Gly Glu Gln Cys Val His Thr Ala Ser Gly Pro Arg Cys Phe Cys 1355 1360 1365 Pro Ser Pro Arg Asp Cys Glu Ser Gly Cys Ala Ser Ser Pro Cys 1370 1375 1380 Gln His Gly Gly Ser Cys His Pro Gln Arg Gln Pro Pro Tyr Tyr 1385 1390 1395 Ser Cys Gln Cys Ala Pro Pro Phe Ser Gly Ser Arg Cys Glu Leu 1400 1405 1410 Tyr Thr Ala Pro Pro Ser Thr Pro Pro Ala Thr Cys Leu Ser Gln 1415 1420 1425 Tyr Cys Ala Asp Lys Ala Arg Asp Gly Val Cys Asp Glu Ala Cys 1430 1435 1440 Asn Ser His Ala Cys Gln Trp Asp Gly Gly Asp Cys Ser Leu Thr 1445 1450 1455 Met Glu Asn Pro Trp Ala Asn Cys Ser Ser Pro Leu Pro Cys Trp 1460 1465 1470 Asp Tyr Ile Asn Asn Gln Cys Asp Glu Leu Cys Asn Thr Val Glu 1475 1480 1485 Cys Leu Phe Asp Asn Phe Glu Cys Gln Gly Asn Ser Lys Thr Cys 1490 1495 1500 Lys Tyr Asp Lys Tyr Cys Ala Asp His Phe Lys Asp Asn His Cys 1505 1510 1515 Asp Gln Gly Cys Asn Ser Glu Glu Cys Gly Trp Asp Gly Leu Asp 1520 1525 1530 Cys Ala Ala Asp Gln Pro Glu Asn Leu Ala Glu Gly Thr Leu Val 1535 1540 1545 Ile Val Val Leu Met Pro Pro Glu Gln Leu Leu Gln Asp Ala Arg 1550 1555 1560 Ser Phe Leu Arg Ala Leu Gly Thr Leu Leu His Thr Asn Leu Arg 1565 1570 1575 Ile Lys Arg Asp Ser Gln Gly Glu Leu Met Val Tyr Pro Tyr Tyr 1580 1585 1590 Gly Glu Lys Ser Ala Ala Met Lys Lys Gln Arg Met Thr Arg Arg 1595 1600 1605 Ser Leu Pro Gly Glu Gln Glu Gln Glu Val Ala Gly Ser Lys Val 1610 1615 1620 Phe Leu Glu Ile Asp Asn Arg Gln Cys Val Gln Asp Ser Asp His 1625 1630 1635 Cys Phe Lys Asn Thr Asp Ala Ala Ala Ala Leu Leu Ala Ser His 1640 1645 1650 Ala Ile Gln Gly Thr Leu Ser Tyr Pro Leu Val Ser Val Val Ser 1655 1660 1665 Glu Ser Leu Thr Pro Glu Arg Thr Gln Leu Leu Tyr Leu Leu Ala 1670 1675 1680 Val Ala Val Val Ile Ile Leu Phe Ile Ile Leu Leu Gly Val Ile 1685 1690 1695 Met Ala Lys Arg Lys Arg Lys His Gly Ser Leu Trp Leu Pro Glu 1700 1705 1710 Gly Phe Thr Leu Arg Arg Asp Ala Ser Asn His Lys Arg Arg Glu 1715 1720 1725 Pro Val Gly Gln Asp Ala Val Gly Leu Lys Asn Leu Ser Val Gln 1730 1735 1740 Val Ser Glu Ala Asn Leu Ile Gly Thr Gly Thr Ser Glu His Trp 1745 1750 1755 Val Asp Asp Glu Gly Pro Gln Pro Lys Lys Val Lys Ala Glu Asp 1760 1765 1770 Glu Ala Leu Leu Ser Glu Glu Asp Asp Pro Ile Asp Arg Arg Pro 1775 1780 1785 Trp Thr Gln Gln His Leu Glu Ala Ala Asp Ile Arg Arg Thr Pro 1790 1795 1800 Ser Leu Ala Leu Thr Pro Pro Gln Ala Glu Gln Glu Val Asp Val 1805 1810 1815 Leu Asp Val Asn Val Arg Gly Pro Asp Gly Cys Thr Pro Leu Met 1820 1825 1830 Leu Ala Ser Leu Arg Gly Gly Ser Ser Asp Leu Ser Asp Glu Asp 1835 1840 1845 Glu Asp Ala Glu Asp Ser Ser Ala Asn Ile Ile Thr Asp Leu Val 1850 1855 1860 Tyr Gln Gly Ala Ser Leu Gln Ala Gln Thr Asp Arg Thr Gly Glu 1865 1870 1875 Met Ala Leu His Leu Ala Ala Arg Tyr Ser Arg Ala Asp Ala Ala 1880 1885 1890 Lys Arg Leu Leu Asp Ala Gly Ala Asp Ala Asn Ala Gln Asp Asn 1895 1900 1905 Met Gly Arg Cys Pro Leu His Ala Ala Val Ala Ala Asp Ala Gln 1910 1915 1920 Gly Val Phe Gln Ile Leu Ile Arg Asn Arg Val Thr Asp Leu Asp 1925 1930 1935 Ala Arg Met Asn Asp Gly Thr Thr Pro Leu Ile Leu Ala Ala Arg 1940 1945 1950 Leu Ala Val Glu Gly Met Val Ala Glu Leu Ile Asn Cys Gln Ala 1955 1960 1965 Asp Val Asn Ala Val Asp Asp His Gly Lys Ser Ala Leu His Trp 1970 1975 1980 Ala Ala Ala Val Asn Asn Val Glu Ala Thr Leu Leu Leu Leu Lys 1985 1990 1995 Asn Gly Ala Asn Arg Asp Met Gln Asp Asn Lys Glu Glu Thr Pro 2000 2005 2010 Leu Phe Leu Ala Ala Arg Glu Gly Ser Tyr Glu Ala Ala Lys Ile 2015 2020 2025 Leu Leu Asp His Phe Ala Asn Arg Asp Ile Thr Asp His Met Asp 2030 2035 2040 Arg Leu Pro Arg Asp Val Ala Arg Asp Arg Met His His Asp Ile 2045 2050 2055 Val Arg Leu Leu Asp Glu Tyr Asn Val Thr Pro Ser Pro Pro Gly 2060 2065 2070 Thr Val Leu Thr Ser Ala Leu Ser Pro Val Ile Cys Gly Pro Asn 2075 2080 2085 Arg Ser Phe Leu Ser Leu Lys His Thr Pro Met Gly Lys Lys Ser 2090 2095 2100 Arg Arg Pro Ser Ala Lys Ser Thr Met Pro Thr Ser Leu Pro Asn 2105 2110 2115 Leu Ala Lys Glu Ala Lys Asp Ala Lys Gly Ser Arg Arg Lys Lys 2120 2125 2130 Ser Leu Ser Glu Lys Val Gln Leu Ser Glu Ser Ser Val Thr Leu 2135 2140 2145 Ser Pro Val Asp Ser Leu Glu Ser Pro His Thr Tyr Val Ser Asp 2150 2155 2160 Thr Thr Ser Ser Pro Met Ile Thr Ser Pro Gly Ile Leu Gln Ala 2165 2170 2175 Ser Pro Asn Pro Met Leu Ala Thr Ala Ala Pro Pro Ala Pro Val 2180 2185 2190 His Ala Gln His Ala Leu Ser Phe Ser Asn Leu His Glu Met Gln 2195 2200 2205 Pro Leu Ala His Gly Ala Ser Thr Val Leu Pro Ser Val Ser Gln 2210 2215 2220 Leu Leu Ser His His His Ile Val Ser Pro Gly Ser Gly Ser Ala 2225 2230 2235 Gly Ser Leu Ser Arg Leu His Pro Val Pro Val Pro Ala Asp Trp 2240 2245 2250 Met Asn Arg Met Glu Val Asn Glu Thr Gln Tyr Asn Glu Met Phe 2255 2260 2265 Gly Met Val Leu Ala Pro Ala Glu Gly Thr His

Pro Gly Ile Ala 2270 2275 2280 Pro Gln Ser Arg Pro Pro Glu Gly Lys His Ile Thr Thr Pro Arg 2285 2290 2295 Glu Pro Leu Pro Pro Ile Val Thr Phe Gln Leu Ile Pro Lys Gly 2300 2305 2310 Ser Ile Ala Gln Pro Ala Gly Ala Pro Gln Pro Gln Ser Thr Cys 2315 2320 2325 Pro Pro Ala Val Ala Gly Pro Leu Pro Thr Met Tyr Gln Ile Pro 2330 2335 2340 Glu Met Ala Arg Leu Pro Ser Val Ala Phe Pro Thr Ala Met Met 2345 2350 2355 Pro Gln Gln Asp Gly Gln Val Ala Gln Thr Ile Leu Pro Ala Tyr 2360 2365 2370 His Pro Phe Pro Ala Ser Val Gly Lys Tyr Pro Thr Pro Pro Ser 2375 2380 2385 Gln His Ser Tyr Ala Ser Ser Asn Ala Ala Glu Arg Thr Pro Ser 2390 2395 2400 His Ser Gly His Leu Gln Gly Glu His Pro Tyr Leu Thr Pro Ser 2405 2410 2415 Pro Glu Ser Pro Asp Gln Trp Ser Ser Ser Ser Pro His Ser Ala 2420 2425 2430 Ser Asp Trp Ser Asp Val Thr Thr Ser Pro Thr Pro Gly Gly Ala 2435 2440 2445 Gly Gly Gly Gln Arg Gly Pro Gly Thr His Met Ser Glu Pro Pro 2450 2455 2460 His Asn Asn Met Gln Val Tyr Ala 2465 2470 3 2319 PRT Homo sapiens 3 Met Gly Leu Gly Ala Arg Gly Arg Arg Arg Arg Arg Arg Leu Met Ala 1 5 10 15 Leu Pro Pro Pro Pro Pro Pro Met Arg Ala Leu Pro Leu Leu Leu Leu 20 25 30 Leu Ala Gly Leu Gly Ala Ala Ala Pro Pro Cys Leu Asp Gly Ser Pro 35 40 45 Cys Ala Asn Gly Gly Arg Cys Thr His Gln Gln Pro Ser Leu Glu Ala 50 55 60 Ala Cys Leu Cys Leu Pro Gly Trp Val Gly Glu Arg Cys Gln Leu Glu 65 70 75 80 Asp Pro Cys His Ser Gly Pro Cys Ala Gly Arg Gly Val Cys Gln Ser 85 90 95 Ser Val Val Ala Gly Thr Ala Arg Phe Ser Cys Arg Cys Leu Arg Gly 100 105 110 Phe Gln Gly Pro Asp Cys Ser Gln Pro Asp Pro Cys Val Ser Arg Pro 115 120 125 Cys Val His Gly Ala Pro Cys Ser Val Gly Pro Asp Gly Arg Phe Ala 130 135 140 Cys Ala Cys Pro Pro Gly Tyr Gln Gly Gln Ser Cys Gln Ser Asp Ile 145 150 155 160 Asp Glu Cys Arg Ser Gly Thr Thr Cys Arg His Gly Gly Thr Cys Leu 165 170 175 Asn Thr Pro Gly Ser Phe Arg Cys Gln Cys Pro Leu Gly Tyr Thr Gly 180 185 190 Leu Leu Cys Glu Asn Pro Val Val Pro Cys Ala Pro Ser Pro Cys Arg 195 200 205 Asn Gly Gly Thr Cys Arg Gln Ser Ser Asp Val Thr Tyr Asp Cys Ala 210 215 220 Cys Leu Pro Gly Phe Glu Gly Gln Asn Cys Glu Val Asn Val Asp Asp 225 230 235 240 Cys Pro Gly His Arg Cys Leu Asn Gly Gly Thr Cys Val Asp Gly Val 245 250 255 Asn Thr Tyr Asn Cys Gln Cys Pro Pro Glu Trp Thr Gly Gln Phe Cys 260 265 270 Thr Glu Asp Val Asp Glu Cys Gln Leu Gln Pro Asn Ala Cys His Asn 275 280 285 Gly Gly Thr Cys Phe Asn Leu Leu Gly Gly His Ser Cys Val Cys Val 290 295 300 Asn Gly Trp Thr Gly Glu Ser Cys Ser Gln Asn Ile Asp Asp Cys Ala 305 310 315 320 Thr Ala Val Cys Phe His Gly Ala Thr Cys His Asp Arg Val Ala Ser 325 330 335 Phe Tyr Cys Ala Cys Pro Met Gly Lys Thr Gly Leu Leu Cys His Leu 340 345 350 Asp Asp Ala Cys Val Ser Asn Pro Cys His Glu Asp Ala Ile Cys Asp 355 360 365 Thr Asn Pro Val Ser Gly Arg Ala Ile Cys Thr Cys Pro Pro Gly Phe 370 375 380 Thr Gly Gly Ala Cys Asp Gln Asp Val Asp Glu Cys Ser Ile Gly Ala 385 390 395 400 Asn Pro Cys Glu His Leu Gly Arg Cys Val Asn Thr Gln Gly Ser Phe 405 410 415 Leu Cys Gln Cys Gly Arg Gly Tyr Thr Gly Pro Arg Cys Glu Thr Asp 420 425 430 Val Asn Glu Cys Leu Ser Gly Pro Cys Arg Asn Gln Ala Thr Cys Leu 435 440 445 Asp Arg Ile Gly Gln Phe Thr Cys Ile Cys Met Ala Gly Phe Thr Gly 450 455 460 Thr Tyr Cys Glu Val Asp Ile Asp Glu Cys Gln Ser Ser Pro Cys Val 465 470 475 480 Asn Gly Gly Val Cys Lys Asp Arg Val Asn Gly Phe Ser Cys Thr Cys 485 490 495 Pro Ser Gly Phe Ser Gly Ser Met Cys Gln Leu Asp Val Asp Glu Cys 500 505 510 Ala Ser Thr Pro Cys Arg Asn Gly Ala Lys Cys Val Asp Gln Pro Asp 515 520 525 Gly Tyr Glu Cys Arg Cys Ala Glu Gly Phe Glu Gly Thr Leu Cys Glu 530 535 540 Arg Asn Val Asp Asp Cys Ser Pro Asp Pro Cys His His Gly Arg Cys 545 550 555 560 Val Asp Gly Ile Ala Ser Phe Ser Cys Ala Cys Ala Pro Gly Tyr Thr 565 570 575 Gly Ile Arg Cys Glu Ser Gln Val Asp Glu Cys Arg Ser Gln Pro Cys 580 585 590 Arg Tyr Gly Gly Lys Cys Leu Asp Leu Val Asp Lys Tyr Leu Cys Arg 595 600 605 Cys Pro Pro Gly Thr Thr Gly Val Asn Cys Glu Val Asn Ile Asp Asp 610 615 620 Cys Ala Ser Asn Pro Cys Thr Phe Gly Val Cys Arg Asp Gly Ile Asn 625 630 635 640 Arg Tyr Asp Cys Val Cys Gln Pro Gly Phe Thr Gly Pro Leu Cys Asn 645 650 655 Val Glu Ile Asn Glu Cys Ala Ser Ser Pro Cys Gly Glu Gly Gly Ser 660 665 670 Cys Val Asp Gly Glu Asn Gly Phe His Cys Leu Cys Pro Pro Gly Ser 675 680 685 Leu Pro Pro Leu Cys Leu Pro Ala Asn His Pro Cys Ala His Lys Pro 690 695 700 Cys Ser His Gly Val Cys His Asp Ala Pro Gly Gly Phe Arg Cys Val 705 710 715 720 Cys Glu Pro Gly Trp Ser Gly Pro Arg Cys Ser Gln Ser Leu Ala Pro 725 730 735 Asp Ala Cys Glu Ser Gln Pro Cys Gln Ala Gly Gly Thr Cys Thr Ser 740 745 750 Asp Gly Ile Gly Phe Arg Cys Thr Cys Ala Pro Gly Phe Gln Gly His 755 760 765 Gln Cys Glu Val Leu Ser Pro Cys Thr Pro Ser Leu Cys Glu His Gly 770 775 780 Gly His Cys Glu Ser Asp Pro Asp Arg Leu Thr Val Cys Ser Cys Pro 785 790 795 800 Pro Gly Trp Gln Gly Pro Arg Cys Gln Gln Asp Val Asp Glu Cys Ala 805 810 815 Gly Ala Ser Pro Cys Gly Pro His Gly Thr Cys Thr Asn Leu Pro Gly 820 825 830 Asn Phe Arg Cys Ile Cys His Arg Gly Tyr Thr Gly Pro Phe Cys Asp 835 840 845 Gln Asp Ile Asp Asp Cys Asp Pro Asn Pro Cys Leu His Gly Gly Ser 850 855 860 Cys Gln Asp Gly Val Gly Ser Phe Ser Cys Ser Cys Leu Asp Gly Phe 865 870 875 880 Ala Gly Pro Arg Cys Ala Arg Asp Val Asp Glu Cys Leu Ser Ser Pro 885 890 895 Cys Gly Pro Gly Thr Cys Thr Asp His Val Ala Ser Phe Thr Cys Ala 900 905 910 Cys Pro Pro Gly Tyr Gly Gly Phe His Cys Glu Ile Asp Leu Pro Asp 915 920 925 Cys Ser Pro Ser Ser Cys Phe Asn Gly Gly Thr Cys Val Asp Gly Val 930 935 940 Ser Ser Phe Ser Cys Leu Cys Arg Pro Gly Tyr Thr Gly Thr His Cys 945 950 955 960 Gln Tyr Glu Ala Asp Pro Cys Phe Ser Arg Pro Cys Leu His Gly Gly 965 970 975 Ile Cys Asn Pro Thr His Pro Gly Phe Glu Cys Thr Cys Arg Glu Gly 980 985 990 Phe Thr Gly Ser Gln Cys Gln Asn Pro Val Asp Trp Cys Ser Gln Ala 995 1000 1005 Pro Cys Gln Asn Gly Gly Arg Cys Val Gln Thr Gly Ala Tyr Cys 1010 1015 1020 Ile Cys Pro Pro Gly Trp Ser Gly Arg Leu Cys Asp Ile Gln Ser 1025 1030 1035 Leu Pro Cys Thr Glu Ala Ala Ala Gln Met Gly Val Arg Leu Glu 1040 1045 1050 Gln Leu Cys Gln Glu Gly Gly Lys Cys Ile Asp Lys Gly Arg Ser 1055 1060 1065 His Tyr Cys Val Cys Pro Glu Gly Arg Thr Gly Ser His Cys Glu 1070 1075 1080 His Glu Val Asp Pro Cys Thr Ala Gln Pro Cys Gln His Gly Gly 1085 1090 1095 Thr Cys Arg Gly Tyr Met Gly Gly Tyr Val Cys Glu Cys Pro Ala 1100 1105 1110 Gly Tyr Ala Gly Asp Ser Cys Glu Asp Asn Ile Asp Glu Cys Ala 1115 1120 1125 Ser Gln Pro Cys Gln Asn Gly Gly Ser Cys Ile Asp Leu Val Ala 1130 1135 1140 Arg Tyr Leu Cys Ser Cys Pro Pro Gly Thr Leu Gly Val Leu Cys 1145 1150 1155 Glu Ile Asn Glu Asp Asp Cys Asp Leu Gly Pro Ser Leu Asp Ser 1160 1165 1170 Gly Val Gln Cys Leu His Asn Gly Thr Cys Val Asp Leu Val Gly 1175 1180 1185 Gly Phe Arg Cys Asn Cys Pro Pro Gly Tyr Thr Gly Leu His Cys 1190 1195 1200 Glu Ala Asp Ile Asn Glu Cys Arg Pro Gly Ala Cys His Ala Ala 1205 1210 1215 His Thr Arg Asp Cys Leu Gln Asp Pro Gly Gly His Phe Arg Cys 1220 1225 1230 Val Cys His Pro Gly Phe Thr Gly Pro Arg Cys Gln Ile Ala Leu 1235 1240 1245 Ser Pro Cys Glu Ser Gln Pro Cys Gln His Gly Gly Gln Cys Arg 1250 1255 1260 His Ser Leu Gly Arg Gly Gly Gly Leu Thr Phe Thr Cys His Cys 1265 1270 1275 Val Pro Pro Phe Trp Gly Leu Arg Cys Glu Arg Val Ala Arg Ser 1280 1285 1290 Cys Arg Glu Leu Gln Cys Pro Val Gly Ile Pro Cys Gln Gln Thr 1295 1300 1305 Ala Arg Gly Pro Arg Cys Ala Cys Pro Pro Gly Leu Ser Gly Pro 1310 1315 1320 Ser Cys Arg Val Ser Arg Ala Ser Pro Ser Gly Ala Thr Asn Ala 1325 1330 1335 Ser Cys Ala Ser Ala Pro Cys Leu His Gly Gly Ser Cys Leu Pro 1340 1345 1350 Val Gln Ser Val Pro Phe Phe Arg Cys Val Cys Ala Pro Gly Trp 1355 1360 1365 Gly Gly Pro Arg Cys Glu Thr Pro Ser Ala Ala Pro Glu Val Pro 1370 1375 1380 Glu Glu Pro Arg Cys Pro Arg Ala Ala Cys Gln Ala Lys Arg Gly 1385 1390 1395 Asp Gln Asn Cys Asp Arg Glu Cys Asn Thr Pro Gly Cys Gly Trp 1400 1405 1410 Asp Gly Gly Asp Cys Ser Leu Asn Val Asp Asp Pro Trp Arg Gln 1415 1420 1425 Cys Glu Ala Leu Gln Cys Trp Arg Leu Phe Asn Asn Ser Arg Cys 1430 1435 1440 Asp Pro Ala Cys Ser Ser Pro Ala Cys Leu Tyr Asp Asn Phe Asp 1445 1450 1455 Cys Tyr Ser Gly Gly Arg Asp Arg Thr Cys Asn Pro Val Tyr Glu 1460 1465 1470 Lys Tyr Cys Ala Asp His Phe Ala Asp Gly Arg Cys Asp Gln Gly 1475 1480 1485 Cys Asn Thr Glu Glu Cys Gly Trp Asp Gly Leu Asp Cys Ala Ser 1490 1495 1500 Glu Val Pro Ala Leu Leu Ala Arg Gly Val Leu Val Leu Thr Val 1505 1510 1515 Leu Leu Pro Pro Glu Glu Leu Leu Arg Ser Ser Ala Asp Phe Leu 1520 1525 1530 Gln Arg Leu Ser Ala Ile Leu Arg Thr Ser Leu Arg Phe Arg Leu 1535 1540 1545 Asp Ala Arg Gly Gln Ala Met Val Phe Pro Tyr His Arg Pro Ser 1550 1555 1560 Pro Gly Ser Glu Ser Arg Val Arg Arg Glu Leu Gly Pro Glu Val 1565 1570 1575 Ile Gly Ser Val Val Met Leu Glu Ile Asp Asn Arg Leu Cys Leu 1580 1585 1590 Gln Ser Ala Glu Asn Asp His Cys Phe Pro Asp Ala Gln Ser Ala 1595 1600 1605 Ala Asp Tyr Leu Gly Ala Leu Ser Ala Val Glu Arg Leu Asp Phe 1610 1615 1620 Pro Tyr Pro Leu Arg Asp Val Arg Gly Glu Pro Leu Glu Ala Pro 1625 1630 1635 Glu Gln Ser Val Pro Leu Leu Pro Leu Leu Val Ala Gly Ala Val 1640 1645 1650 Phe Leu Leu Ile Ile Phe Ile Leu Gly Val Met Val Ala Arg Arg 1655 1660 1665 Lys Arg Glu His Ser Thr Leu Trp Phe Pro Glu Gly Phe Ala Leu 1670 1675 1680 His Lys Asp Ile Ala Ala Gly His Lys Gly Arg Arg Glu Pro Val 1685 1690 1695 Gly Gln Asp Ala Leu Gly Met Lys Asn Met Ala Lys Gly Glu Ser 1700 1705 1710 Leu Met Gly Glu Val Val Thr Asp Leu Asn Asp Ser Glu Cys Pro 1715 1720 1725 Glu Ala Lys Arg Leu Lys Val Glu Glu Pro Gly Met Gly Ala Glu 1730 1735 1740 Glu Pro Glu Asp Cys Arg Gln Trp Thr Gln His His Leu Val Ala 1745 1750 1755 Ala Asp Ile Arg Val Ala Pro Ala Thr Ala Leu Thr Pro Pro Gln 1760 1765 1770 Gly Asp Ala Asp Ala Asp Gly Val Asp Val Asn Val Arg Gly Pro 1775 1780 1785 Asp Gly Phe Thr Pro Leu Met Leu Ala Ser Phe Cys Gly Gly Ala 1790 1795 1800 Leu Glu Pro Met Pro Ala Glu Glu Asp Glu Ala Asp Asp Thr Ser 1805 1810 1815 Ala Ser Ile Ile Ser Asp Leu Ile Cys Gln Gly Ala Gln Leu Gly 1820 1825 1830 Ala Arg Thr Asp Arg Thr Gly Glu Thr Ala Leu His Leu Ala Ala 1835 1840 1845 Arg Tyr Ala Arg Ala Asp Ala Ala Lys Arg Leu Leu Asp Ala Gly 1850 1855 1860 Ala Asp Thr Asn Ala Gln Asp His Ser Gly Arg Thr Pro Leu His 1865 1870 1875 Thr Ala Val Thr Ala Asp Ala Gln Gly Val Phe Gln Ile Leu Ile 1880 1885 1890 Arg Asn Arg Ser Thr Asp Leu Asp Ala Arg Met Ala Asp Gly Ser 1895 1900 1905 Thr Ala Leu Ile Leu Ala Ala Arg Leu Ala Val Glu Gly Met Val 1910 1915 1920 Glu Glu Leu Ile Ala Ser His Ala Asp Val Asn Ala Val Asp Glu 1925 1930 1935 Leu Gly Lys Ser Ala Leu His Trp Ala Ala Ala Val Asn Asn Val 1940 1945 1950 Glu Ala Thr Leu Ala Leu Leu Lys Asn Gly Ala Asn Lys Asp Met 1955 1960 1965 Gln Asp Ser Lys Glu Glu Thr Pro Leu Phe Leu Ala Ala Arg Glu 1970 1975 1980 Gly Ser Tyr Glu Ala Ala Lys Leu Leu Leu Asp His Leu Ala Asn 1985 1990 1995 Arg Glu Ile Thr Asp His Leu Asp Arg Leu Pro Arg Asp Val Ala 2000 2005 2010 Gln Glu Arg Leu His Gln Asp Ile Val Arg Leu Leu Asp Gln Pro 2015 2020 2025 Ser Gly Pro Arg Ser Pro Ser Gly Pro His Gly Leu Gly Pro Leu 2030 2035 2040 Leu Cys Pro Pro Gly Ala Phe Leu Pro Gly Leu Lys Ala Val Gln 2045 2050 2055 Ser Gly Thr Lys Lys Ser Arg Arg Pro Pro Gly Lys Thr Gly Leu 2060 2065 2070 Gly Pro Gln Gly Thr Arg Gly Arg Gly Lys Lys Leu Thr Leu Ala 2075 2080 2085 Cys Pro Gly Pro Leu Ala Asp Ser Ser Val Thr Leu Ser Pro Val 2090 2095 2100 Asp Ser Leu Asp Ser Pro Arg Pro Phe Ser Gly Pro Pro Ala Ser 2105 2110 2115 Pro Gly Gly Phe Pro Leu Glu Gly Pro Tyr Ala Thr Thr Ala Thr 2120 2125 2130 Ala Val Ser Leu Ala Gln Leu Gly Ala Ser Arg Ala Gly Pro Leu 2135 2140 2145 Gly Arg Gln Pro Pro Gly Gly Cys Val Leu Ser Phe Gly Leu Leu 2150 2155 2160 Asn Pro Val Ala Val Pro Leu Asp Trp Ala Arg Leu Pro Pro Pro 2165 2170 2175 Ala Pro Pro Gly Pro Ser Phe Leu Leu Pro Leu Ala Pro Gly Pro 2180 2185 2190 Gln Leu Leu Asn Pro Gly Ala Pro Val Ser Pro Gln Glu Arg Pro 2195 2200 2205 Pro Pro Tyr Leu Ala Ala Pro Gly His Gly Glu Glu Tyr Pro Ala 2210

2215 2220 Ala Gly Thr Arg Ser Ser Pro Thr Lys Ala Arg Phe Leu Arg Val 2225 2230 2235 Pro Ser Glu His Pro Tyr Leu Thr Pro Ser Pro Glu Ser Pro Glu 2240 2245 2250 His Trp Ala Ser Pro Ser Pro Pro Ser Leu Ser Asp Trp Ser Asp 2255 2260 2265 Ser Thr Pro Ser Pro Ala Thr Ala Thr Asn Ala Thr Ala Ser Gly 2270 2275 2280 Ala Leu Pro Ala Gln Pro His Pro Ile Ser Val Pro Ser Leu Pro 2285 2290 2295 Gln Ser Gln Thr Gln Leu Gly Pro Gln Pro Glu Val Thr Pro Lys 2300 2305 2310 Arg Gln Val Met Ala Glx 2315 4 2002 PRT Homo sapiens 4 Met Gln Pro Pro Ser Leu Leu Leu Leu Leu Leu Leu Leu Leu Leu Cys 1 5 10 15 Val Ser Val Val Arg Pro Arg Gly Leu Leu Cys Gly Ser Phe Pro Glu 20 25 30 Pro Cys Ala Asn Gly Gly Thr Cys Leu Ser Leu Ser Leu Gly Gln Gly 35 40 45 Thr Cys Gln Cys Ala Pro Gly Phe Leu Gly Glu Thr Cys Gln Phe Pro 50 55 60 Asp Pro Cys Gln Asn Ala Gln Leu Cys Gln Asn Gly Gly Ser Cys Gln 65 70 75 80 Ala Leu Leu Pro Ala Pro Leu Gly Leu Pro Ser Ser Pro Ser Pro Leu 85 90 95 Thr Pro Ser Phe Leu Cys Thr Cys Leu Pro Gly Phe Thr Gly Glu Arg 100 105 110 Cys Gln Ala Lys Leu Glu Asp Pro Cys Pro Pro Ser Phe Cys Ser Lys 115 120 125 Arg Gly Arg Cys His Ile Gln Ala Ser Gly Arg Pro Gln Cys Ser Cys 130 135 140 Met Pro Gly Trp Thr Gly Glu Gln Cys Gln Leu Arg Asp Phe Cys Ser 145 150 155 160 Ala Asn Pro Cys Val Asn Gly Gly Val Cys Leu Ala Thr Tyr Pro Gln 165 170 175 Ile Gln Cys His Cys Pro Pro Gly Phe Glu Gly His Ala Cys Glu Arg 180 185 190 Asp Val Asn Glu Cys Phe Gln Asp Pro Gly Pro Cys Pro Lys Gly Thr 195 200 205 Ser Cys His Asn Thr Leu Gly Ser Phe Gln Cys Leu Cys Pro Val Gly 210 215 220 Gln Glu Gly Pro Arg Cys Glu Leu Arg Ala Gly Pro Cys Pro Pro Arg 225 230 235 240 Gly Cys Ser Asn Gly Gly Thr Cys Gln Leu Met Pro Glu Lys Asp Ser 245 250 255 Thr Phe His Leu Cys Leu Cys Pro Pro Gly Phe Ile Gly Pro Gly Cys 260 265 270 Glu Val Asn Pro Asp Asn Cys Val Ser His Gln Cys Gln Asn Gly Gly 275 280 285 Thr Cys Gln Asp Gly Leu Asp Thr Tyr Thr Cys Leu Cys Pro Glu Thr 290 295 300 Trp Thr Gly Trp Asp Cys Ser Glu Asp Val Asp Glu Cys Glu Ala Gln 305 310 315 320 Gly Pro Pro His Cys Arg Asn Gly Gly Thr Cys Gln Asn Ser Ala Gly 325 330 335 Ser Phe His Cys Val Cys Val Ser Gly Trp Gly Gly Thr Ser Cys Glu 340 345 350 Glu Asn Leu Asp Asp Cys Ile Ala Ala Thr Cys Ala Pro Gly Ser Thr 355 360 365 Cys Ile Asp Arg Val Gly Ser Phe Ser Cys Leu Cys Pro Pro Gly Arg 370 375 380 Thr Gly Leu Leu Cys His Leu Glu Asp Met Cys Leu Ser Gln Pro Cys 385 390 395 400 His Gly Asp Ala Gln Cys Ser Thr Asn Pro Leu Thr Gly Ser Thr Leu 405 410 415 Cys Leu Cys Gln Pro Gly Tyr Ser Gly Pro Thr Cys His Gln Asp Leu 420 425 430 Asp Glu Cys Leu Met Ala Gln Gln Gly Pro Ser Pro Cys Glu His Gly 435 440 445 Gly Ser Cys Leu Asn Thr Pro Gly Ser Phe Asn Cys Leu Cys Pro Pro 450 455 460 Gly Tyr Thr Gly Ser Arg Cys Glu Ala Asp His Asn Glu Cys Leu Ser 465 470 475 480 Gln Pro Cys His Pro Gly Ser Thr Cys Leu Asp Leu Leu Ala Thr Phe 485 490 495 His Cys Leu Cys Pro Pro Gly Leu Glu Gly Gln Leu Cys Glu Val Glu 500 505 510 Thr Asn Glu Cys Ala Ser Ala Pro Cys Leu Asn His Ala Asp Cys His 515 520 525 Asp Leu Leu Asn Gly Phe Gln Cys Ile Cys Leu Pro Gly Phe Ser Gly 530 535 540 Thr Arg Cys Glu Glu Asp Ile Asp Glu Cys Arg Ser Ser Pro Cys Ala 545 550 555 560 Asn Gly Gly Gln Cys Gln Asp Gln Pro Gly Ala Phe His Cys Lys Cys 565 570 575 Leu Pro Gly Phe Glu Gly Pro Arg Cys Gln Thr Glu Val Asp Glu Cys 580 585 590 Leu Ser Asp Pro Cys Pro Val Gly Ala Ser Cys Leu Asp Leu Pro Gly 595 600 605 Ala Phe Phe Cys Leu Cys Pro Ser Gly Phe Thr Gly Gln Leu Cys Glu 610 615 620 Val Pro Leu Cys Ala Pro Asn Leu Cys Gln Pro Lys Gln Ile Cys Lys 625 630 635 640 Asp Gln Lys Asp Lys Ala Asn Cys Leu Cys Pro Asp Gly Ser Pro Gly 645 650 655 Cys Ala Pro Pro Glu Asp Asn Cys Thr Cys His His Gly His Cys Gln 660 665 670 Arg Ser Ser Cys Val Cys Asp Val Gly Trp Thr Gly Pro Glu Cys Glu 675 680 685 Ala Glu Leu Gly Gly Cys Ile Ser Ala Pro Cys Ala His Gly Gly Thr 690 695 700 Cys Tyr Pro Gln Pro Ser Gly Tyr Asn Cys Thr Cys Pro Thr Gly Tyr 705 710 715 720 Thr Gly Pro Thr Cys Ser Glu Glu Met Thr Ala Cys His Ser Gly Pro 725 730 735 Cys Leu Asn Gly Gly Ser Cys Asn Pro Ser Pro Gly Gly Tyr Tyr Cys 740 745 750 Thr Cys Pro Pro Ser His Thr Gly Pro Gln Cys Gln Thr Ser Thr Asp 755 760 765 Tyr Cys Val Ser Ala Pro Cys Phe Asn Gly Gly Thr Cys Val Asn Arg 770 775 780 Pro Gly Thr Phe Ser Cys Leu Cys Ala Met Gly Phe Gln Gly Pro Arg 785 790 795 800 Cys Glu Gly Lys Leu Arg Pro Ser Cys Ala Asp Ser Pro Cys Arg Asn 805 810 815 Arg Ala Thr Cys Gln Asp Ser Pro Gln Gly Pro Arg Cys Leu Cys Pro 820 825 830 Thr Gly Tyr Thr Gly Gly Ser Cys Gln Thr Leu Met Asp Leu Cys Ala 835 840 845 Gln Lys Pro Cys Pro Arg Asn Ser His Cys Leu Gln Thr Gly Pro Ser 850 855 860 Phe His Cys Leu Cys Leu Gln Gly Trp Thr Gly Pro Leu Cys Asn Leu 865 870 875 880 Pro Leu Ser Ser Cys Gln Lys Ala Ala Leu Ser Gln Gly Ile Asp Val 885 890 895 Ser Ser Leu Cys His Asn Gly Gly Leu Cys Val Asp Ser Gly Pro Ser 900 905 910 Tyr Phe Cys His Cys Pro Pro Gly Phe Gln Gly Ser Leu Cys Gln Asp 915 920 925 His Val Asn Pro Cys Glu Ser Arg Pro Cys Gln Asn Gly Ala Thr Cys 930 935 940 Met Ala Gln Pro Ser Gly Tyr Leu Cys Gln Cys Ala Pro Gly Tyr Asp 945 950 955 960 Gly Gln Asn Cys Ser Lys Glu Leu Asp Ala Cys Gln Ser Gln Pro Cys 965 970 975 His Asn His Gly Thr Cys Thr Pro Lys Pro Gly Gly Phe His Cys Ala 980 985 990 Cys Pro Pro Gly Phe Val Gly Leu Arg Cys Glu Gly Asp Val Asp Glu 995 1000 1005 Cys Leu Asp Gln Pro Cys His Pro Thr Gly Thr Ala Ala Cys His 1010 1015 1020 Ser Leu Ala Asn Ala Phe Tyr Cys Gln Cys Leu Pro Gly His Thr 1025 1030 1035 Gly Gln Trp Cys Glu Val Glu Ile Asp Pro Cys His Ser Gln Pro 1040 1045 1050 Cys Phe His Gly Gly Thr Cys Glu Ala Thr Ala Gly Ser Pro Leu 1055 1060 1065 Gly Phe Ile Cys His Cys Pro Lys Gly Phe Glu Gly Pro Thr Cys 1070 1075 1080 Ser His Arg Ala Pro Ser Cys Gly Phe His His Cys His His Gly 1085 1090 1095 Gly Leu Cys Leu Pro Ser Pro Lys Pro Gly Phe Pro Pro Arg Cys 1100 1105 1110 Ala Cys Leu Ser Gly Tyr Gly Gly Pro Asp Cys Leu Thr Pro Pro 1115 1120 1125 Ala Pro Lys Gly Cys Gly Pro Pro Ser Pro Cys Leu Tyr Asn Gly 1130 1135 1140 Ser Cys Ser Glu Thr Thr Gly Leu Gly Gly Pro Gly Phe Arg Cys 1145 1150 1155 Ser Cys Pro His Ser Ser Pro Gly Pro Arg Cys Gln Lys Pro Gly 1160 1165 1170 Ala Lys Gly Cys Glu Gly Arg Ser Gly Asp Gly Ala Cys Asp Ala 1175 1180 1185 Gly Cys Ser Gly Pro Gly Gly Asn Trp Asp Gly Gly Asp Cys Ser 1190 1195 1200 Leu Gly Val Pro Asp Pro Trp Lys Gly Cys Pro Ser His Ser Arg 1205 1210 1215 Cys Trp Leu Leu Phe Arg Asp Gly Gln Cys His Pro Gln Cys Asp 1220 1225 1230 Ser Glu Glu Cys Leu Phe Asp Gly Tyr Asp Cys Glu Thr Pro Pro 1235 1240 1245 Ala Cys Thr Pro Ala Tyr Asp Gln Tyr Cys His Asp His Phe His 1250 1255 1260 Asn Gly His Cys Glu Lys Gly Cys Asn Thr Ala Glu Cys Gly Trp 1265 1270 1275 Asp Gly Gly Asp Cys Arg Pro Glu Asp Gly Asp Pro Glu Trp Gly 1280 1285 1290 Pro Ser Leu Ala Leu Leu Val Val Leu Ser Pro Pro Ala Leu Asp 1295 1300 1305 Gln Gln Leu Phe Ala Leu Ala Arg Val Leu Ser Leu Thr Leu Arg 1310 1315 1320 Val Gly Leu Trp Val Arg Lys Asp Arg Asp Gly Arg Asp Met Val 1325 1330 1335 Tyr Pro Tyr Pro Gly Ala Arg Ala Glu Glu Lys Leu Gly Gly Thr 1340 1345 1350 Arg Asp Pro Thr Tyr Gln Glu Arg Ala Ala Pro Gln Thr Gln Pro 1355 1360 1365 Leu Gly Lys Glu Thr Asp Ser Leu Ser Ala Gly Phe Val Val Val 1370 1375 1380 Met Gly Val Asp Leu Ser Arg Cys Gly Pro Asp His Pro Ala Ser 1385 1390 1395 Arg Cys Pro Trp Asp Pro Gly Leu Leu Leu Arg Phe Leu Ala Ala 1400 1405 1410 Met Ala Ala Val Gly Ala Leu Glu Pro Leu Leu Pro Gly Pro Leu 1415 1420 1425 Leu Ala Val His Pro His Ala Gly Thr Ala Pro Pro Ala Asn Gln 1430 1435 1440 Leu Pro Trp Pro Val Leu Cys Ser Pro Val Ala Gly Val Ile Leu 1445 1450 1455 Leu Ala Leu Gly Ala Leu Leu Val Leu Gln Leu Ile Arg Arg Arg 1460 1465 1470 Arg Arg Glu His Gly Ala Leu Trp Leu Pro Pro Gly Phe Thr Arg 1475 1480 1485 Arg Pro Arg Thr Gln Ser Ala Pro His Arg Arg Arg Pro Pro Leu 1490 1495 1500 Gly Glu Asp Ser Ile Gly Leu Lys Ala Leu Lys Pro Lys Ala Glu 1505 1510 1515 Val Asp Glu Asp Gly Val Val Met Cys Ser Gly Pro Glu Glu Gly 1520 1525 1530 Glu Glu Val Gly Gln Ala Glu Glu Thr Gly Pro Pro Ser Thr Cys 1535 1540 1545 Gln Leu Trp Ser Leu Ser Gly Gly Cys Gly Ala Leu Pro Gln Ala 1550 1555 1560 Ala Met Leu Thr Pro Pro Gln Glu Ser Glu Met Glu Ala Pro Asp 1565 1570 1575 Leu Asp Thr Arg Gly Pro Asp Gly Val Thr Pro Leu Met Ser Ala 1580 1585 1590 Val Cys Cys Gly Glu Val Gln Ser Gly Thr Phe Gln Gly Ala Trp 1595 1600 1605 Leu Gly Cys Pro Glu Pro Trp Glu Pro Leu Leu Asp Gly Gly Ala 1610 1615 1620 Cys Pro Gln Ala His Thr Val Gly Thr Gly Glu Thr Pro Leu His 1625 1630 1635 Leu Ala Ala Arg Phe Ser Arg Pro Thr Ala Ala Arg Arg Leu Leu 1640 1645 1650 Glu Ala Gly Ala Asn Pro Asn Gln Pro Asp Arg Ala Gly Arg Thr 1655 1660 1665 Pro Leu His Ala Ala Val Ala Ala Asp Ala Arg Glu Val Cys Gln 1670 1675 1680 Leu Leu Leu Arg Ser Arg Gln Thr Ala Val Asp Ala Arg Thr Glu 1685 1690 1695 Asp Gly Thr Thr Pro Leu Met Leu Ala Ala Arg Leu Ala Val Glu 1700 1705 1710 Asp Leu Val Glu Glu Leu Ile Ala Ala Gln Ala Asp Val Gly Ala 1715 1720 1725 Arg Asp Lys Trp Gly Lys Thr Ala Leu His Trp Ala Ala Ala Val 1730 1735 1740 Asn Asn Ala Arg Ala Ala Arg Ser Leu Leu Gln Ala Gly Ala Asp 1745 1750 1755 Lys Asp Ala Gln Asp Asn Arg Glu Gln Thr Pro Leu Phe Leu Ala 1760 1765 1770 Ala Arg Glu Gly Ala Val Glu Val Ala Gln Leu Leu Leu Gly Leu 1775 1780 1785 Gly Ala Ala Arg Glu Leu Arg Asp Gln Ala Gly Leu Ala Pro Ala 1790 1795 1800 Asp Val Ala His Gln Arg Asn His Trp Asp Leu Leu Thr Leu Leu 1805 1810 1815 Glu Gly Ala Gly Pro Pro Glu Ala Arg His Lys Ala Thr Pro Gly 1820 1825 1830 Arg Glu Ala Gly Pro Phe Pro Arg Ala Arg Thr Val Ser Val Ser 1835 1840 1845 Val Pro Pro His Gly Gly Gly Ala Leu Pro Arg Cys Arg Thr Leu 1850 1855 1860 Ser Ala Gly Ala Gly Pro Arg Gly Gly Gly Ala Cys Leu Gln Ala 1865 1870 1875 Arg Thr Trp Ser Val Asp Leu Ala Ala Arg Gly Gly Gly Ala Tyr 1880 1885 1890 Ser His Cys Arg Ser Leu Ser Gly Val Gly Ala Gly Gly Gly Pro 1895 1900 1905 Thr Pro Arg Gly Arg Arg Phe Ser Ala Gly Met Arg Gly Pro Arg 1910 1915 1920 Pro Asn Pro Ala Ile Met Arg Gly Arg Tyr Gly Val Ala Ala Gly 1925 1930 1935 Arg Gly Gly Arg Val Ser Thr Asp Asp Trp Pro Cys Asp Trp Val 1940 1945 1950 Ala Leu Gly Ala Cys Gly Ser Ala Ser Asn Ile Pro Ile Pro Pro 1955 1960 1965 Pro Cys Leu Thr Pro Ser Pro Glu Arg Gly Ser Pro Gln Leu Asp 1970 1975 1980 Cys Gly Pro Pro Ala Leu Gln Glu Met Pro Ile Asn Gln Gly Gly 1985 1990 1995 Glu Gly Lys Lys 2000 5 48 PRT Homo sapiens 5 Ala Ala Gly Thr Leu Val Val Val Val Leu Met Pro Pro Glu Gln Leu 1 5 10 15 Arg Asn Ser Ser Phe His Phe Leu Arg Glu Leu Ser Arg Val Leu His 20 25 30 Thr Asn Val Val Phe Lys Arg Asp Ala His Gly Gln Gln Met Ile Phe 35 40 45 6 48 PRT Rattus rattus 6 Ala Ala Gly Thr Leu Val Leu Val Val Leu Leu Pro Pro Asp Gln Leu 1 5 10 15 Arg Asn Asn Ser Phe His Phe Leu Arg Asp Val Ser His Val Leu His 20 25 30 Thr Asn Val Val Phe Lys Arg Asp Ala His Gly Gln Gln Met Ile Phe 35 40 45 7 48 PRT Mus musculus 7 Ala Ala Gly Thr Leu Val Leu Val Val Leu Leu Pro Pro Asp Gln Leu 1 5 10 15 Arg Asn Asn Ser Phe His Phe Leu Arg Glu Leu Ser His Val Leu His 20 25 30 Thr Asn Val Val Phe Lys Arg Asp Ala His Gly Gln Gln Met Ile Phe 35 40 45 8 48 PRT Gallus gallus 8 Ala Asp Gly Thr Leu Val Val Val Val Leu Ile Thr Pro Glu Asn Leu 1 5 10 15 Lys Asn Asn Ser Phe Asn Phe Leu Arg Glu Leu Ser Arg Val Leu His 20 25 30 Thr Asn Val Val Phe Lys Lys Asn Ala His Gly Gln Gln Met Ile Phe 35 40 45 9 48 PRT Xenopus laevis 9 Ala Glu Gly Thr Leu Val Leu Val Val Leu Met Pro Pro Glu Arg Leu 1 5 10 15 Lys Asn Asn Ser Val Asn Phe Leu Arg Glu Leu Ser Arg Val Leu His 20 25 30 Thr Asn Val Val Phe Lys Lys Asp Ala His Gly Gln Gln Met Ile Phe 35 40 45 10 48 PRT Danio rerio 10 Ala Val Gly Leu Leu Val Val Val Val His Ile His Pro Asp Gln Leu 1 5 10 15 Arg Asn Asn Ser Phe Gly Phe Leu Arg Glu Leu Ser Arg Val Leu His 20 25 30 Thr Asn Val Val Phe Arg Arg Asp Ala His Gly Gln Gln Met Ile Phe 35 40 45 11 48 PRT artificial sequence consensus sequence misc_feature (2)..(2) Xaa=Val, Glu, Asp or

Ala misc_feature (4)..(4) Xaa=Leu or Thr misc_feature (7)..(7) Xaa=Val or Leu misc_feature (10)..(10) Xaa=His or Leu misc_feature (11)..(11) Xaa=Ile, Met or Leu misc_feature (12)..(12) Xaa=His, Pro or Thr misc_feature (14)..(14) Xaa=Asp or Glu misc_feature (15)..(15) Xaa=Gln, Arg or Asn misc_feature (17)..(17) Xaa=Arg or Lys misc_feature (19)..(19) Xaa=Asn or Ser misc_feature (21)..(21) Xaa=Phe or Val misc_feature (22)..(22) Xaa=Gly, Asn or His misc_feature (26)..(26) Xaa=Asp or Glu misc_feature (27)..(27) Xaa=Leu or Val misc_feature (29)..(29) Xaa=Arg or His misc_feature (38)..(38) Xaa=Arg or Lys misc_feature (39)..(39) Xaa=Arg or Lys 11 Ala Xaa Gly Xaa Leu Val Xaa Val Val Xaa Xaa Xaa Pro Xaa Xaa Leu 1 5 10 15 Xaa Asn Xaa Ser Xaa Xaa Phe Leu Arg Xaa Xaa Ser Xaa Val Leu His 20 25 30 Thr Asn Val Val Phe Xaa Xaa Asp Ala His Gly Gln Gln Met Ile Phe 35 40 45 12 48 PRT Homo sapiens 12 Ala Ala Gly Thr Leu Val Val Val Val Leu Met Pro Pro Glu Gln Leu 1 5 10 15 Arg Asn Ser Ser Phe His Phe Leu Arg Glu Leu Ser Arg Val Pro His 20 25 30 Thr Asn Val Val Phe Lys Arg Asp Ala His Gly Gln Gln Met Ile Phe 35 40 45 13 48 PRT Homo sapiens 13 Ala Ala Gly Thr Leu Val Val Val Val Leu Met Pro Pro Glu Gln Leu 1 5 10 15 Arg Asn Ser Ser Phe His Phe Pro Arg Glu Leu Ser Arg Val Leu His 20 25 30 Thr Asn Val Val Phe Lys Arg Asp Ala His Gly Gln Gln Met Ile Phe 35 40 45 14 48 PRT Homo sapiens 14 Ala Ala Gly Thr Leu Val Val Val Val Leu Met Pro Pro Glu Gln Leu 1 5 10 15 Arg Asn Ser Ser Phe His Phe Pro Arg Glu Leu Ser Arg Val Leu His 20 25 30 Thr Asn Val Val Phe Lys Arg Val Ala His Gly Gln Gln Met Ile Phe 35 40 45 15 48 PRT Homo sapiens 15 Ala Ala Gly Thr Leu Val Val Val Val Leu Met Pro Pro Glu Gln Leu 1 5 10 15 Arg Asn Ser Ser Phe His Phe Leu Arg Glu Leu Ser Pro Val Leu His 20 25 30 Thr Asn Val Val Phe Lys Arg Asp Ala His Gly Gln Gln Met Ile Phe 35 40 45 16 48 PRT Homo sapiens 16 Ala Ala Gly Thr Pro Val Val Val Val Leu Met Pro Pro Glu Gln Leu 1 5 10 15 Arg Asn Ser Ser Phe His Phe Leu Arg Glu Leu Ser Arg Val Leu His 20 25 30 Thr Asn Val Val Phe Lys Arg Asp Ala His Gly Gln Gln Met Ile Phe 35 40 45 17 47 PRT Homo sapiens 17 Ala Ala Gly Thr Leu Val Val Val Leu Met Pro Pro Glu Gln Leu Arg 1 5 10 15 Asn Ser Ser Phe His Phe Leu Arg Glu Leu Ser Arg Val Leu His Thr 20 25 30 Asn Val Val Phe Lys Arg Asp Ala His Gly Gln Gln Met Ile Phe 35 40 45 18 48 PRT Homo sapiens 18 Ala Ala Gly Thr Leu Val Val Val Val Leu Met Pro Pro Glu Gln Leu 1 5 10 15 Arg Asn Ser Ser Phe His Ser Leu Arg Glu Leu Ser Arg Val Leu His 20 25 30 Thr Asn Val Val Phe Lys Arg Asp Ala His Gly Gln Gln Met Ile Phe 35 40 45 19 48 PRT Homo sapiens 19 Ala Ala Gly Thr Leu Val Val Val Val Leu Met Pro Pro Glu Gln Pro 1 5 10 15 Arg Asn Ser Ser Phe His Phe Leu Arg Glu Leu Ser Arg Val Leu His 20 25 30 Thr Asn Val Val Phe Lys Arg Asp Ala His Gly Gln Gln Met Ile Phe 35 40 45 20 48 PRT Homo sapiens 20 Ala Ala Gly Thr Leu Val Val Val Val Leu Met Pro Pro Glu Gln Leu 1 5 10 15 Arg Asn Ser Ser Phe His Phe Leu Arg Glu Leu Ser Arg Val Leu His 20 25 30 Thr Asn Val Val Phe Lys Ser Asp Ala His Gly Gln Gln Met Ile Phe 35 40 45 21 47 PRT Homo sapiens 21 Ala Ala Gly Thr Leu Val Val Val Val Leu Met Pro Pro Glu Gln Leu 1 5 10 15 Arg Asn Ser Ser Phe His Phe Leu Arg Glu Leu Ser Arg Val Leu His 20 25 30 Thr Asn Val Phe Lys Arg Asp Ala His Gly Gln Gln Met Ile Phe 35 40 45 22 46 PRT Homo sapiens 22 Ala Ala Gly Thr Leu Val Val Val Val Leu Met Pro Pro Glu Gln Leu 1 5 10 15 Arg Asn Ser Ser Phe His Phe Leu Arg Glu Leu Ser Arg Val Leu His 20 25 30 Thr Asn Val Val Phe Lys Thr His Gly Gln Gln Met Ile Phe 35 40 45 23 53 PRT Homo sapiens 23 Ala Ala Gly Thr Leu Val Val Val Val Leu Met Pro Pro Glu Gln Leu 1 5 10 15 Arg Asn Ser Ser Phe His Phe Leu Arg Glu Leu Ser Arg Val Leu His 20 25 30 Thr Asn Val Val Phe Lys Arg Asp Ala His Gly Gln Gln Met Ile Phe 35 40 45 Pro Pro Gly Gly Gly 50 24 53 PRT Homo sapiens 24 Ala Ala Gly Thr Leu Val Val Val Val Leu Met Pro Pro Ala Gln Gln 1 5 10 15 Leu Arg Asn Ser Ser Phe His Phe Leu Arg Glu Leu Ser Arg Val Leu 20 25 30 His Thr Asn Val Val Phe Lys Arg Asp Ala His Gly Gln Gln Met Ile 35 40 45 Phe Pro Tyr Tyr Gly 50 25 53 PRT Homo sapiens 25 Ala Ala Gly Thr Leu Val Val Val Val Leu Met Pro Pro Glu Gln Leu 1 5 10 15 Arg Asn Ser Ser Phe His Phe Leu Arg Glu Leu Ser Arg Val Leu His 20 25 30 Thr Asn Val Val Leu Gly Arg Arg Asp Ala His Gly Gln Gln Met Ile 35 40 45 Phe Pro Tyr Tyr Gly 50 26 53 PRT Homo sapiens 26 Ala Ala Gly Thr Leu Val Val Val Val Leu Met Pro Pro Glu Gln Leu 1 5 10 15 Arg Asn Ser Ser Phe His Leu Ala Leu Arg Glu Leu Ser Arg Val Leu 20 25 30 His Thr Asn Val Val Phe Lys Arg Asp Ala His Gly Gln Gln Met Ile 35 40 45 Phe Pro Tyr Tyr Gly 50 27 55 PRT Homo sapiens 27 Ala Ala Gly Thr Leu Val Val Val Val Leu Met Pro Pro Glu Gln Leu 1 5 10 15 Arg Asn Ser Ser Phe His Phe Leu Arg Glu Lys Glu Asp Leu Ser Arg 20 25 30 Val Leu His Thr Asn Val Val Phe Lys Arg Asp Ala His Gly Gln Gln 35 40 45 Met Ile Phe Pro Tyr Tyr Gly 50 55 28 56 PRT Homo sapiens 28 Ala Ala Gly Thr Leu Val Val Val Val Leu Met Pro Pro Glu Gln Leu 1 5 10 15 Arg Asn Ser Ser Phe His Phe Leu Arg Glu Leu Ser Arg Val Leu His 20 25 30 Thr Asn Val Val Phe Lys Ser Met Pro Pro Arg Asp Ala His Gly Gln 35 40 45 Gln Met Ile Phe Pro Tyr Tyr Gly 50 55 29 53 PRT Homo sapiens 29 Ala Ala Gly Thr Leu Val Val Val Val Leu Met Pro Pro Glu Gln Leu 1 5 10 15 Arg Asn Ser Ser Phe His Thr Ala Leu Arg Glu Leu Ser Arg Val Leu 20 25 30 His Thr Asn Val Val Phe Lys Arg Asp Ala His Gly Gln Gln Met Ile 35 40 45 Phe Pro Tyr Tyr Gly 50 30 88 PRT Homo sapiens 30 Trp Leu Ser Arg Leu Gln Ser Gly Met Val Pro Asn Gln Tyr Asn Pro 1 5 10 15 Leu Arg Gly Ser Val Ala Pro Gly Pro Leu Ser Thr Gln Ala Pro Ser 20 25 30 Leu Gln His Gly Met Val Gly Pro Leu His Ser Ser Leu Ala Ala Ser 35 40 45 Ala Leu Ser Gln Met Met Ser Tyr Gln Gly Leu Pro Ser Thr Arg Leu 50 55 60 Ala Thr Gln Pro His Leu Val Gln Thr Gln Gln Val Gln Pro Gln Asn 65 70 75 80 Leu Gln Met Gln Gln Gln Asn Leu 85 31 84 PRT Homo sapiens 31 Trp Leu Ser Arg Leu Gln Ser Gly Met Val Pro Asn Gln Tyr Asn Pro 1 5 10 15 Leu Arg Gly Ser Val Ala Pro Gly Pro Leu Ser Thr Gln Ala Pro Ser 20 25 30 Leu Gln His Gly Met Val Gly Pro Leu His Ser Ser Leu Ala Ala Ser 35 40 45 Ala Leu Ser Gln Met Met Ser Tyr Gln Gly Leu Pro Ser Thr Arg Leu 50 55 60 Ala Thr Gln Pro His Leu Val Gln Thr Gln Gln Val Gln Pro Gln Asn 65 70 75 80 Leu Gln Met Gln 32 83 PRT Homo sapiens 32 Trp Leu Ser Arg Leu Gln Ser Gly Met Val Pro Asn Gln Tyr Asn Pro 1 5 10 15 Leu Arg Gly Ser Val Ala Pro Gly Pro Leu Ser Thr Gln Ala Pro Ser 20 25 30 Leu Gln His Gly Met Val Gly Pro Leu His Ser Ser Leu Ala Ala Ser 35 40 45 Ala Leu Ser Gln Met Met Ser Tyr Gln Gly Leu Pro Ser Thr Arg Leu 50 55 60 Ala Thr Gln Pro His Leu Val Gln Thr Gln Gln Val Gln Pro Gln Asn 65 70 75 80 Leu Gln Met 33 31 PRT Homo sapiens 33 Trp Leu Ser Arg Leu Gln Ser Gly Met Val Pro Asn Gln Tyr Asn Pro 1 5 10 15 Leu Arg Gly Ser Val Ala Pro Gly Pro Leu Ser Thr Gln Ala Pro 20 25 30 34 166 PRT Homo sapiens 34 Trp Leu Ser Arg Leu Gln Ser Gly Met Val Pro Asn Gln Tyr Asn Pro 1 5 10 15 Leu Arg Gly Ser Val Ala Pro Gly Pro Leu Ser Thr Gln Ala Pro Ser 20 25 30 Leu Gln His Gly Met Val Gly Pro Leu His Ser Ser Leu Ala Ala Ser 35 40 45 Ala Leu Ser Gln Met Met Ser Tyr Gln Gly Leu Pro Ser Thr Arg Leu 50 55 60 Ala Thr Gln Pro His Leu Val Gln Thr Gln Gln Val Gln Pro Gln Asn 65 70 75 80 Leu Gln Met Gln Gln Gln Asn Leu Gln Pro Ala Asn Ile Gln Gln Gln 85 90 95 Gln Ser Leu Gln Pro Pro Pro Pro Pro Pro Gln Pro His Leu Gly Val 100 105 110 Ser Ser Ala Ala Ser Gly His Leu Gly Arg Ser Phe Leu Ser Gly Glu 115 120 125 Pro Ser Gln Ala Asp Val Gln Pro Leu Gly Pro Ser Ser Leu Ala Val 130 135 140 His Thr Ile Leu Pro Gln Glu Ser Pro Ala Leu Pro Thr Ser Leu Pro 145 150 155 160 Ser Ser Leu Val Pro Pro 165 35 167 PRT Homo sapiens 35 Trp Leu Ser Arg Leu Gln Ser Gly Met Val Pro Asn Gln Tyr Asn Pro 1 5 10 15 Leu Arg Gly Ser Val Ala Pro Gly Pro Leu Ser Thr Gln Ala Pro Ser 20 25 30 Leu Gln His Gly Met Val Gly Pro Leu His Ser Ser Leu Ala Ala Ser 35 40 45 Ala Leu Ser Gln Met Met Ser Tyr Gln Gly Leu Pro Ser Thr Arg Leu 50 55 60 Ala Thr Gln Pro His Leu Val Gln Thr Gln Gln Val Gln Pro Gln Asn 65 70 75 80 Leu Gln Met Gln Gln Gln Asn Leu Gln Pro Ala Asn Ile Gln Gln Gln 85 90 95 Gln Ser Leu Gln Pro Pro Pro Pro Pro Pro Gln Pro His Leu Gly Val 100 105 110 Ser Ser Ala Ala Ser Gly His Leu Gly Arg Ser Phe Leu Ser Gly Glu 115 120 125 Pro Ser Gln Ala Asp Val Gln Pro Leu Gly Pro Ser Ser Leu Ala Val 130 135 140 His Thr Ile Leu Pro Gln Glu Ser Pro Ala Leu Pro Thr Ser Leu Pro 145 150 155 160 Ser Ser Leu Val Ala His Pro 165 36 167 PRT Homo sapiens 36 Trp Leu Ser Arg Leu Gln Ser Gly Met Val Pro Asn Gln Tyr Asn Pro 1 5 10 15 Leu Arg Gly Ser Val Ala Pro Gly Pro Leu Ser Thr Gln Ala Pro Ser 20 25 30 Leu Gln His Gly Met Val Gly Pro Leu His Ser Ser Leu Ala Ala Ser 35 40 45 Ala Leu Ser Gln Met Met Ser Tyr Gln Gly Leu Pro Ser Thr Arg Leu 50 55 60 Ala Thr Gln Pro His Leu Val Gln Thr Gln Gln Val Gln Pro Gln Asn 65 70 75 80 Leu Gln Met Gln Gln Gln Asn Leu Gln Pro Ala Asn Ile Gln Gln Gln 85 90 95 Gln Ser Leu Gln Pro Pro Pro Pro Pro Pro Gln Pro His Leu Gly Val 100 105 110 Ser Ser Ala Ala Ser Gly His Leu Gly Arg Ser Phe Leu Ser Gly Glu 115 120 125 Pro Ser Gln Ala Asp Val Gln Pro Leu Gly Pro Ser Ser Leu Ala Val 130 135 140 His Thr Ile Leu Pro Gln Glu Ser Pro Ala Leu Pro Thr Ser Gly Cys 145 150 155 160 His Pro Arg Trp Ser His Pro 165 37 156 PRT Homo sapiens 37 Trp Leu Ser Arg Leu Gln Ser Gly Met Val Pro Asn Gln Tyr Asn Pro 1 5 10 15 Leu Arg Gly Ser Val Ala Pro Gly Pro Leu Ser Thr Gln Ala Pro Ser 20 25 30 Leu Gln His Gly Met Val Gly Pro Leu His Ser Ser Leu Ala Ala Ser 35 40 45 Ala Leu Ser Gln Met Met Ser Tyr Gln Gly Leu Pro Ser Thr Arg Leu 50 55 60 Ala Thr Gln Pro His Leu Val Gln Thr Gln Gln Val Gln Pro Gln Asn 65 70 75 80 Leu Gln Met Gln Gln Gln Asn Leu Gln Pro Ala Asn Ile Gln Gln Gln 85 90 95 Gln Ser Leu Gln Pro Pro Pro Pro Pro Pro Gln Pro His Leu Gly Val 100 105 110 Ser Ser Ala Ala Ser Gly His Leu Gly Arg Ser Phe Leu Ser Gly Glu 115 120 125 Pro Ser Gln Ala Asp Val Gln Pro Leu Gly Pro Ser Ser Leu Ala Val 130 135 140 His Thr Ile Leu Pro Gln Glu Ser Pro Ala Leu Pro 145 150 155 38 166 PRT Homo sapiens 38 Trp Leu Ser Arg Leu Gln Ser Gly Met Val Pro Asn Gln Tyr Asn Pro 1 5 10 15 Leu Arg Gly Ser Val Ala Pro Gly Pro Leu Ser Thr Gln Ala Pro Ser 20 25 30 Leu Gln His Gly Met Val Gly Pro Leu His Ser Ser Leu Ala Ala Ser 35 40 45 Ala Leu Ser Gln Met Met Ser Tyr Gln Gly Leu Pro Ser Thr Arg Leu 50 55 60 Ala Thr Gln Pro His Leu Val Gln Thr Gln Gln Val Gln Pro Gln Asn 65 70 75 80 Leu Gln Met Gln Gln Gln Asn Leu Gln Pro Ala Asn Ile Gln Gln Gln 85 90 95 Gln Ser Leu Gln Pro Pro Pro Pro Pro Pro Gln Pro His Leu Gly Val 100 105 110 Ser Ser Ala Ala Ser Gly His Leu Gly Arg Ser Phe Leu Ser Gly Glu 115 120 125 Pro Ser Gln Ala Asp Val Gln Pro Leu Gly Pro Ser Ser Leu Ala Val 130 135 140 His Thr Ile Leu Pro Gln Glu Ser Pro Pro Cys Pro Arg Arg Cys His 145 150 155 160 Pro Arg Trp Ser His Pro 165 39 150 PRT Homo sapiens 39 Trp Leu Ser Arg Leu Gln Ser Gly Met Val Pro Asn Gln Tyr Asn Pro 1 5 10 15 Leu Arg Gly Ser Val Ala Pro Gly Pro Leu Ser Thr Gln Ala Pro Ser 20 25 30 Leu Gln His Gly Met Val Gly Pro Leu His Ser Ser Leu Ala Ala Ser 35 40 45 Ala Leu Ser Gln Met Met Ser Tyr Gln Gly Leu Pro Ser Thr Arg Leu 50 55 60 Ala Thr Gln Pro His Leu Val Gln Thr Gln Gln Val Gln Pro Gln Asn 65 70 75 80 Leu Gln Met Gln Gln Gln Asn Leu Gln Pro Ala Asn Ile Gln Gln Gln 85 90 95 Gln Ser Leu Gln Pro Pro Pro Pro Pro Pro Gln Pro His Leu Gly Val 100 105 110 Ser Ser Ala Ala Ser Gly His Leu Gly Arg Ser Phe Leu Ser Gly Glu 115 120 125 Pro Ser Gln Ala Asp Val Gln Pro Leu Gly Pro Ser Ser Leu Ala Val 130 135 140 His Thr Ile Leu Pro Gln 145 150 40 170 PRT Homo sapiens 40 Trp Leu Ser Arg Leu Gln Ser Gly Met Val Pro Asn Gln Tyr Asn Pro 1 5 10 15 Leu Arg Gly Ser Val Ala Pro Gly Pro Leu Ser Thr Gln Ala Pro Ser 20 25 30 Leu Gln His Gly Met Val Gly Pro Leu His Ser Ser Leu Ala Ala Ser 35 40 45 Ala Leu Ser Gln Met Met Ser Tyr Gln Gly Leu Pro Ser Thr Arg Leu 50 55 60 Ala Thr Gln Pro His Leu Val Gln Thr Gln

Gln Val Gln Pro Gln Asn 65 70 75 80 Leu Gln Met Gln Gln Gln Asn Leu Gln Pro Ala Asn Ile Gln Gln Gln 85 90 95 Gln Ser Leu Gln Pro Pro Pro Pro Pro Pro Gln Pro His Leu Gly Val 100 105 110 Ser Ser Ala Ala Ser Gly His Leu Gly Arg Ser Phe Leu Ser Gly Glu 115 120 125 Pro Ser Gln Ala Asp Leu Ser Arg Asp Leu Cys Ser His Trp Ala Pro 130 135 140 Ala Ala Trp Arg Cys Thr Leu Phe Cys Pro Arg Arg Ala Pro Pro Cys 145 150 155 160 Pro Arg Arg Cys His Pro Arg Trp Ser His 165 170 41 167 PRT Homo sapiens 41 Trp Leu Ser Arg Leu Gln Ser Gly Met Val Pro Asn Gln Tyr Asn Pro 1 5 10 15 Leu Arg Gly Ser Val Ala Pro Gly Pro Leu Ser Thr Gln Ala Pro Ser 20 25 30 Leu Gln His Gly Met Val Gly Pro Leu His Ser Ser Leu Ala Ala Ser 35 40 45 Ala Leu Ser Gln Met Met Ser Tyr Gln Gly Leu Pro Ser Thr Arg Leu 50 55 60 Ala Thr Gln Pro His Leu Val Gln Thr Gln Gln Val Gln Pro Gln Asn 65 70 75 80 Leu Gln Met Gln Gln Gln Asn Leu Gln Pro Ala Asn Ile Gln Gln Gln 85 90 95 Gln Ser Leu Gln Pro Pro Pro Pro Pro Pro Gln Pro His Leu Gly Val 100 105 110 Ser Ser Ala Ala Ser Gly His Leu Gly Arg Ser Phe Leu Ser Gly Glu 115 120 125 Pro Ser Gln Ala Asp Cys Cys Ser His Trp Ala Pro Ala Ala Trp Arg 130 135 140 Cys Thr Leu Phe Cys Pro Arg Arg Ala Pro Pro Cys Pro Arg Arg Cys 145 150 155 160 His Pro Arg Trp Ser His Pro 165 42 170 PRT Homo sapiens 42 Trp Leu Ser Arg Leu Gln Ser Gly Met Val Pro Asn Gln Tyr Asn Pro 1 5 10 15 Leu Arg Gly Ser Val Ala Pro Gly Pro Leu Ser Thr Gln Ala Pro Ser 20 25 30 Leu Gln His Gly Met Val Gly Pro Leu His Ser Ser Leu Ala Ala Ser 35 40 45 Ala Leu Ser Gln Met Met Ser Tyr Gln Gly Leu Pro Ser Thr Arg Leu 50 55 60 Ala Thr Gln Pro His Leu Val Gln Thr Gln Gln Val Gln Pro Gln Asn 65 70 75 80 Leu Gln Met Gln Gln Gln Asn Leu Gln Pro Ala Asn Ile Gln Gln Gln 85 90 95 Gln Ser Leu Gln Pro Pro Pro Pro Pro Pro Gln Pro His Leu Gly Val 100 105 110 Ser Ser Ala Ala Ser Gly His Leu Gly Arg Ser Phe Leu Ser Gly Glu 115 120 125 Pro Ser Gln Ala Glu Gly Arg Gly Arg Cys Ser His Trp Ala Pro Ala 130 135 140 Ala Trp Arg Cys Thr Leu Phe Cys Pro Arg Arg Ala Pro Pro Cys Pro 145 150 155 160 Arg Arg Cys His Pro Arg Trp Ser His Pro 165 170 43 169 PRT Homo sapiens 43 Trp Leu Ser Arg Leu Gln Ser Gly Met Val Pro Asn Gln Tyr Asn Pro 1 5 10 15 Leu Arg Gly Ser Val Ala Pro Gly Pro Leu Ser Thr Gln Ala Pro Ser 20 25 30 Leu Gln His Gly Met Val Gly Pro Leu His Ser Ser Leu Ala Ala Ser 35 40 45 Ala Leu Ser Gln Met Met Ser Tyr Gln Gly Leu Pro Ser Thr Arg Leu 50 55 60 Ala Thr Gln Pro His Leu Val Gln Thr Gln Gln Val Gln Pro Gln Asn 65 70 75 80 Leu Gln Met Gln Gln Gln Asn Leu Gln Pro Ala Asn Ile Gln Gln Gln 85 90 95 Gln Ser Leu Gln Pro Pro Pro Pro Pro Pro Gln Pro His Leu Gly Val 100 105 110 Ser Ser Ala Ala Ser Gly His Leu Gly Arg Ser Phe Leu Ser Gly Glu 115 120 125 Pro Ser Gln Gly Gly Gly Gly Met Gly Ser His Trp Ala Pro Ala Ala 130 135 140 Trp Arg Cys Thr Leu Phe Cys Pro Arg Arg Ala Pro Pro Cys Pro Arg 145 150 155 160 Arg Cys His Pro Arg Trp Ser His Pro 165 44 167 PRT Homo sapiens 44 Trp Leu Ser Arg Leu Gln Ser Gly Met Val Pro Asn Gln Tyr Asn Pro 1 5 10 15 Leu Arg Gly Ser Val Ala Pro Gly Pro Leu Ser Thr Gln Ala Pro Ser 20 25 30 Leu Gln His Gly Met Val Gly Pro Leu His Ser Ser Leu Ala Ala Ser 35 40 45 Ala Leu Ser Gln Met Met Ser Tyr Gln Gly Leu Pro Ser Thr Arg Leu 50 55 60 Ala Thr Gln Pro His Leu Val Gln Thr Gln Gln Val Gln Pro Gln Asn 65 70 75 80 Leu Gln Met Gln Gln Gln Asn Leu Gln Pro Ala Asn Ile Gln Gln Gln 85 90 95 Gln Ser Leu Gln Pro Pro Pro Pro Pro Pro Gln Pro His Leu Gly Val 100 105 110 Ser Ser Ala Ala Ser Gly His Leu Gly Arg Ser Phe Leu Ser Gly Glu 115 120 125 Glu Arg Ala Arg Gln Thr Cys Ser His Trp Ala Pro Ala Ala Trp Arg 130 135 140 Cys Thr Leu Phe Cys Pro Arg Arg Ala Pro Pro Cys Pro Arg Arg Cys 145 150 155 160 His Pro Arg Trp Ser His Pro 165 45 168 PRT Homo sapiens 45 Trp Leu Ser Arg Leu Gln Ser Gly Met Val Pro Asn Gln Tyr Asn Pro 1 5 10 15 Leu Arg Gly Ser Val Ala Pro Gly Pro Leu Ser Thr Gln Ala Pro Ser 20 25 30 Leu Gln His Gly Met Val Gly Pro Leu His Ser Ser Leu Ala Ala Ser 35 40 45 Ala Leu Ser Gln Met Met Ser Tyr Gln Gly Leu Pro Ser Thr Arg Leu 50 55 60 Ala Thr Gln Pro His Leu Val Gln Thr Gln Gln Val Gln Pro Gln Asn 65 70 75 80 Leu Gln Met Gln Gln Gln Asn Leu Gln Pro Ala Asn Ile Gln Gln Gln 85 90 95 Gln Ser Leu Gln Pro Pro Pro Pro Pro Pro Gln Pro His Leu Gly Val 100 105 110 Ser Ser Ala Ala Ser Gly His Leu Gly Arg Ser Phe Leu Ser Gly Glu 115 120 125 Arg Val Arg Ala Arg Gln Thr Cys Ser His Trp Ala Pro Ala Ala Trp 130 135 140 Arg Cys Thr Leu Phe Cys Pro Arg Arg Ala Pro Pro Cys Pro Arg Arg 145 150 155 160 Cys His Pro Arg Trp Ser His Pro 165 46 128 PRT Homo sapiens 46 Trp Leu Ser Arg Leu Gln Ser Gly Met Val Pro Asn Gln Tyr Asn Pro 1 5 10 15 Leu Arg Gly Ser Val Ala Pro Gly Pro Leu Ser Thr Gln Ala Pro Ser 20 25 30 Leu Gln His Gly Met Val Gly Pro Leu His Ser Ser Leu Ala Ala Ser 35 40 45 Ala Leu Ser Gln Met Met Ser Tyr Gln Gly Leu Pro Ser Thr Arg Leu 50 55 60 Ala Thr Gln Pro His Leu Val Gln Thr Gln Gln Val Gln Pro Gln Asn 65 70 75 80 Leu Gln Met Gln Gln Gln Asn Leu Gln Pro Ala Asn Ile Gln Gln Gln 85 90 95 Gln Ser Leu Gln Pro Pro Pro Pro Pro Pro Gln Pro His Leu Gly Val 100 105 110 Ser Ser Ala Ala Ser Gly His Leu Gly Arg Ser Phe Leu Ser Gly Glu 115 120 125 47 126 PRT Homo sapiens 47 Trp Leu Ser Arg Leu Gln Ser Gly Met Val Pro Asn Gln Tyr Asn Pro 1 5 10 15 Leu Arg Gly Ser Val Ala Pro Gly Pro Leu Ser Thr Gln Ala Pro Ser 20 25 30 Leu Gln His Gly Met Val Gly Pro Leu His Ser Ser Leu Ala Ala Ser 35 40 45 Ala Leu Ser Gln Met Met Ser Tyr Gln Gly Leu Pro Ser Thr Arg Leu 50 55 60 Ala Thr Gln Pro His Leu Val Gln Thr Gln Gln Val Gln Pro Gln Asn 65 70 75 80 Leu Gln Met Gln Gln Gln Asn Leu Gln Pro Ala Asn Ile Gln Gln Gln 85 90 95 Gln Ser Leu Gln Pro Pro Pro Pro Pro Pro Gln Pro His Leu Gly Val 100 105 110 Ser Ser Ala Ala Ser Gly His Leu Gly Arg Ser Ala Pro Ser 115 120 125 48 112 PRT Homo sapiens 48 Trp Leu Ser Arg Leu Gln Ser Gly Met Val Pro Asn Gln Tyr Asn Pro 1 5 10 15 Leu Arg Gly Ser Val Ala Pro Gly Pro Leu Ser Thr Gln Ala Pro Ser 20 25 30 Leu Gln His Gly Met Val Gly Pro Leu His Ser Ser Leu Ala Ala Ser 35 40 45 Ala Leu Ser Gln Met Met Ser Tyr Gln Gly Leu Pro Ser Thr Arg Leu 50 55 60 Ala Thr Gln Pro His Leu Val Gln Thr Gln Gln Val Gln Pro Gln Asn 65 70 75 80 Leu Gln Met Gln Gln Gln Asn Leu Gln Pro Ala Asn Ile Gln Gln Gln 85 90 95 Gln Ser Leu Gln Pro Pro Pro Pro Pro Pro Gln Pro His Leu Ala Ala 100 105 110 49 113 PRT Homo sapiens 49 Trp Leu Ser Arg Leu Gln Ser Gly Met Val Pro Asn Gln Tyr Asn Pro 1 5 10 15 Leu Arg Gly Ser Val Ala Pro Gly Pro Leu Ser Thr Gln Ala Pro Ser 20 25 30 Leu Gln His Gly Met Val Gly Pro Leu His Ser Ser Leu Ala Ala Ser 35 40 45 Ala Leu Ser Gln Met Met Ser Tyr Gln Gly Leu Pro Ser Thr Arg Leu 50 55 60 Ala Thr Gln Pro His Leu Val Gln Thr Gln Gln Val Gln Pro Gln Asn 65 70 75 80 Leu Gln Met Gln Gln Gln Asn Leu Gln Pro Ala Asn Ile Gln Gln Gln 85 90 95 Gln Ser Leu Gln Pro Arg His His His His His Ser Arg Thr Leu Val 100 105 110 Ala 50 96 PRT Homo sapiens 50 Trp Leu Ser Arg Leu Gln Ser Gly Met Val Pro Asn Gln Tyr Asn Pro 1 5 10 15 Leu Arg Gly Ser Val Ala Pro Gly Pro Leu Ser Thr Gln Ala Pro Ser 20 25 30 Leu Gln His Gly Met Val Gly Pro Leu His Ser Ser Leu Ala Ala Ser 35 40 45 Ala Leu Ser Gln Met Met Ser Tyr Gln Gly Leu Pro Ser Thr Arg Leu 50 55 60 Ala Thr Gln Pro His Leu Val Gln Thr Gln Gln Val Gln Pro Gln Asn 65 70 75 80 Leu Gln Met Gln Gln Gln Asn Leu Gln Pro Ala Asn Ile Gln Gln Gln 85 90 95 51 93 PRT Homo sapiens 51 Trp Leu Ser Arg Leu Gln Ser Gly Met Val Pro Asn Gln Tyr Asn Pro 1 5 10 15 Leu Arg Gly Ser Val Ala Pro Gly Pro Leu Ser Thr Gln Ala Pro Ser 20 25 30 Leu Gln His Gly Met Val Gly Pro Leu His Ser Ser Leu Ala Ala Ser 35 40 45 Ala Leu Ser Gln Met Met Ser Tyr Gln Gly Leu Pro Ser Thr Arg Leu 50 55 60 Ala Thr Gln Pro His Leu Val Gln Thr Gln Gln Val Gln Pro Gln Asn 65 70 75 80 Leu Gln Met Gln Gln Gln Asn Leu Gln Pro Ala Asn Ile 85 90 52 275 PRT Homo sapiens 52 Trp Leu Ser Arg Leu Gln Ser Gly Met Val Pro Asn Gln Tyr Asn Pro 1 5 10 15 Leu Arg Gly Ser Val Ala Pro Gly Pro Leu Ser Thr Gln Ala Pro Ser 20 25 30 Leu Gln His Gly Met Val Gly Pro Leu His Ser Ser Leu Ala Ala Ser 35 40 45 Ala Leu Ser Gln Met Met Ser Tyr Gln Gly Leu Pro Ser Thr Arg Leu 50 55 60 Ala Thr Gln Pro His Leu Val Gln Thr Gln Gln Val Gln Pro Gln Asn 65 70 75 80 Leu Gln Met Gln Gln Gln Asn Leu Gln Pro Ala Asn Ile Gln Gln Gln 85 90 95 Gln Ser Leu Gln Pro Pro Pro Pro Pro Pro Gln Pro His Leu Gly Val 100 105 110 Ser Ser Ala Ala Ser Gly His Leu Gly Arg Ser Phe Leu Ser Gly Glu 115 120 125 Pro Ser Gln Ala Asp Val Gln Pro Leu Gly Pro Ser Ser Leu Ala Val 130 135 140 His Thr Ile Leu Pro Gln Glu Ser Pro Ala Leu Pro Thr Ser Leu Pro 145 150 155 160 Ser Ser Leu Val Pro Pro Val Thr Ala Ala Gln Phe Leu Thr Pro Pro 165 170 175 Ser Gln His Ser Tyr Ser Ser Pro Val Asp Asn Thr Pro Ser His Gln 180 185 190 Leu Gln Val Pro Glu His Pro Phe Leu Thr Pro Ser Pro Asp Ser Gly 195 200 205 Pro Ala Arg Pro Arg Ile Pro Thr Ser Pro Thr Gly Pro Arg Ala Ser 210 215 220 Pro Ala Leu Pro Pro Ala Cys Ser Pro Arg Ser Pro Ala Phe Arg Arg 225 230 235 240 Pro Ser Ser Lys Arg Arg Ala Pro Arg Asp Pro Gly Phe Leu Ser Gln 245 250 255 Ala Phe Gly Arg Leu Cys Ala Leu Cys Gly Cys Gln Gly Arg Pro Glu 260 265 270 Glu Pro Phe 275 53 207 PRT Homo sapiens 53 Trp Leu Ser Arg Leu Gln Ser Gly Met Val Pro Asn Gln Tyr Asn Pro 1 5 10 15 Leu Arg Gly Ser Val Ala Pro Gly Pro Leu Ser Thr Gln Ala Pro Ser 20 25 30 Leu Gln His Gly Met Val Gly Pro Leu His Ser Ser Leu Ala Ala Ser 35 40 45 Ala Leu Ser Gln Met Met Ser Tyr Gln Gly Leu Pro Ser Thr Arg Leu 50 55 60 Ala Thr Gln Pro His Leu Val Gln Thr Gln Gln Val Gln Pro Gln Asn 65 70 75 80 Leu Gln Met Gln Gln Gln Asn Leu Gln Pro Ala Asn Ile Gln Gln Gln 85 90 95 Gln Ser Leu Gln Pro Pro Pro Pro Pro Pro Gln Pro His Leu Gly Val 100 105 110 Ser Ser Ala Ala Ser Gly His Leu Gly Arg Ser Phe Leu Ser Gly Glu 115 120 125 Pro Ser Gln Ala Asp Val Gln Pro Leu Gly Pro Ser Ser Leu Ala Val 130 135 140 His Thr Ile Leu Pro Gln Glu Ser Pro Ala Leu Pro Thr Ser Leu Pro 145 150 155 160 Ser Ser Leu Val Pro Pro Val Thr Ala Ala Gln Phe Leu Thr Pro Pro 165 170 175 Ser Gln His Ser Tyr Ser Ser Pro Val Asp Asn Thr Pro Ser His Gln 180 185 190 Leu Gln Val Pro Glu His Pro Phe Leu Thr Pro Ser Ser Val Pro 195 200 205 54 207 PRT Homo sapiens 54 Trp Leu Ser Arg Leu Gln Ser Gly Met Val Pro Asn Gln Tyr Asn Pro 1 5 10 15 Leu Arg Gly Ser Val Ala Pro Gly Pro Leu Ser Thr Gln Ala Pro Ser 20 25 30 Leu Gln His Gly Met Val Gly Pro Leu His Ser Ser Leu Ala Ala Ser 35 40 45 Ala Leu Ser Gln Met Met Ser Tyr Gln Gly Leu Pro Ser Thr Arg Leu 50 55 60 Ala Thr Gln Pro His Leu Val Gln Thr Gln Gln Val Gln Pro Gln Asn 65 70 75 80 Leu Gln Met Gln Gln Gln Asn Leu Gln Pro Ala Asn Ile Gln Gln Gln 85 90 95 Gln Ser Leu Gln Pro Pro Pro Pro Pro Pro Gln Pro His Leu Gly Val 100 105 110 Ser Ser Ala Ala Ser Gly His Leu Gly Arg Ser Phe Leu Ser Gly Glu 115 120 125 Pro Ser Gln Ala Asp Val Gln Pro Leu Gly Pro Ser Ser Leu Ala Val 130 135 140 His Thr Ile Leu Pro Gln Glu Ser Pro Ala Leu Pro Thr Ser Leu Pro 145 150 155 160 Ser Ser Leu Val Pro Pro Val Thr Ala Ala Gln Phe Leu Thr Pro Pro 165 170 175 Ser Gln His Ser Tyr Ser Ser Pro Val Asp Asn Thr Pro Ser His Gln 180 185 190 Leu Gln Val Pro Glu His Pro Phe Leu Thr Pro Ser Arg Val Pro 195 200 205 55 205 PRT Homo sapiens 55 Trp Leu Ser Arg Leu Gln Ser Gly Met Val Pro Asn Gln Tyr Asn Pro 1 5 10 15 Leu Arg Gly Ser Val Ala Pro Gly Pro Leu Ser Thr Gln Ala Pro Ser 20 25 30 Leu Gln His Gly Met Val Gly Pro Leu His Ser Ser Leu Ala Ala Ser 35 40 45 Ala Leu Ser Gln Met Met Ser Tyr Gln Gly Leu Pro Ser Thr Arg Leu 50 55 60 Ala Thr Gln Pro His Leu Val Gln Thr Gln Gln Val Gln Pro Gln Asn 65 70 75 80 Leu Gln Met Gln Gln Gln Asn Leu Gln Pro Ala Asn Ile Gln Gln Gln 85 90 95 Gln Ser Leu Gln Pro Pro Pro

Pro Pro Pro Gln Pro His Leu Gly Val 100 105 110 Ser Ser Ala Ala Ser Gly His Leu Gly Arg Ser Phe Leu Ser Gly Glu 115 120 125 Pro Ser Gln Ala Asp Val Gln Pro Leu Gly Pro Ser Ser Leu Ala Val 130 135 140 His Thr Ile Leu Pro Gln Glu Ser Pro Ala Leu Pro Thr Ser Leu Pro 145 150 155 160 Ser Ser Leu Val Pro Pro Val Thr Ala Ala Gln Phe Leu Thr Pro Pro 165 170 175 Ser Gln His Ser Tyr Ser Ser Pro Val Asp Asn Thr Pro Ser His Gln 180 185 190 Leu Gln Val Pro Glu His Pro Phe Leu Thr Pro Val Pro 195 200 205 56 199 PRT Homo sapiens 56 Trp Leu Ser Arg Leu Gln Ser Gly Met Val Pro Asn Gln Tyr Asn Pro 1 5 10 15 Leu Arg Gly Ser Val Ala Pro Gly Pro Leu Ser Thr Gln Ala Pro Ser 20 25 30 Leu Gln His Gly Met Val Gly Pro Leu His Ser Ser Leu Ala Ala Ser 35 40 45 Ala Leu Ser Gln Met Met Ser Tyr Gln Gly Leu Pro Ser Thr Arg Leu 50 55 60 Ala Thr Gln Pro His Leu Val Gln Thr Gln Gln Val Gln Pro Gln Asn 65 70 75 80 Leu Gln Met Gln Gln Gln Asn Leu Gln Pro Ala Asn Ile Gln Gln Gln 85 90 95 Gln Ser Leu Gln Pro Pro Pro Pro Pro Pro Gln Pro His Leu Gly Val 100 105 110 Ser Ser Ala Ala Ser Gly His Leu Gly Arg Ser Phe Leu Ser Gly Glu 115 120 125 Pro Ser Gln Ala Asp Val Gln Pro Leu Gly Pro Ser Ser Leu Ala Val 130 135 140 His Thr Ile Leu Pro Gln Glu Ser Pro Ala Leu Pro Thr Ser Leu Pro 145 150 155 160 Ser Ser Leu Val Pro Pro Val Thr Ala Ala Gln Phe Leu Thr Pro Pro 165 170 175 Ser Gln His Ser Tyr Ser Ser Pro Val Asp Asn Thr Pro Ser His Gln 180 185 190 Leu Gln Val Pro Glu Tyr Pro 195 57 196 PRT Homo sapiens 57 Trp Leu Ser Arg Leu Gln Ser Gly Met Val Pro Asn Gln Tyr Asn Pro 1 5 10 15 Leu Arg Gly Ser Val Ala Pro Gly Pro Leu Ser Thr Gln Ala Pro Ser 20 25 30 Leu Gln His Gly Met Val Gly Pro Leu His Ser Ser Leu Ala Ala Ser 35 40 45 Ala Leu Ser Gln Met Met Ser Tyr Gln Gly Leu Pro Ser Thr Arg Leu 50 55 60 Ala Thr Gln Pro His Leu Val Gln Thr Gln Gln Val Gln Pro Gln Asn 65 70 75 80 Leu Gln Met Gln Gln Gln Asn Leu Gln Pro Ala Asn Ile Gln Gln Gln 85 90 95 Gln Ser Leu Gln Pro Pro Pro Pro Pro Pro Gln Pro His Leu Gly Val 100 105 110 Ser Ser Ala Ala Ser Gly His Leu Gly Arg Ser Phe Leu Ser Gly Glu 115 120 125 Pro Ser Gln Ala Asp Val Gln Pro Leu Gly Pro Ser Ser Leu Ala Val 130 135 140 His Thr Ile Leu Pro Gln Glu Ser Pro Ala Leu Pro Thr Ser Leu Pro 145 150 155 160 Ser Ser Leu Val Pro Pro Val Thr Ala Ala Gln Phe Leu Thr Pro Pro 165 170 175 Ser Gln His Ser Tyr Ser Ser Pro Val Asp Asn Thr Pro Ser His Gln 180 185 190 Leu Gln Val Pro 195 58 195 PRT Homo sapiens 58 Trp Leu Ser Arg Leu Gln Ser Gly Met Val Pro Asn Gln Tyr Asn Pro 1 5 10 15 Leu Arg Gly Ser Val Ala Pro Gly Pro Leu Ser Thr Gln Ala Pro Ser 20 25 30 Leu Gln His Gly Met Val Gly Pro Leu His Ser Ser Leu Ala Ala Ser 35 40 45 Ala Leu Ser Gln Met Met Ser Tyr Gln Gly Leu Pro Ser Thr Arg Leu 50 55 60 Ala Thr Gln Pro His Leu Val Gln Thr Gln Gln Val Gln Pro Gln Asn 65 70 75 80 Leu Gln Met Gln Gln Gln Asn Leu Gln Pro Ala Asn Ile Gln Gln Gln 85 90 95 Gln Ser Leu Gln Pro Pro Pro Pro Pro Pro Gln Pro His Leu Gly Val 100 105 110 Ser Ser Ala Ala Ser Gly His Leu Gly Arg Ser Phe Leu Ser Gly Glu 115 120 125 Pro Ser Gln Ala Asp Val Gln Pro Leu Gly Pro Ser Ser Leu Ala Val 130 135 140 His Thr Ile Leu Pro Gln Glu Ser Pro Ala Leu Pro Thr Ser Leu Pro 145 150 155 160 Ser Ser Leu Val Pro Pro Val Thr Ala Ala Gln Phe Leu Thr Pro Pro 165 170 175 Ser Gln His Ser Tyr Ser Ser Gly Arg Gln His Pro Gln Pro Pro Ala 180 185 190 Thr Gly Ala 195 59 182 PRT Homo sapiens 59 Trp Leu Ser Arg Leu Gln Ser Gly Met Val Pro Asn Gln Tyr Asn Pro 1 5 10 15 Leu Arg Gly Ser Val Ala Pro Gly Pro Leu Ser Thr Gln Ala Pro Ser 20 25 30 Leu Gln His Gly Met Val Gly Pro Leu His Ser Ser Leu Ala Ala Ser 35 40 45 Ala Leu Ser Gln Met Met Ser Tyr Gln Gly Leu Pro Ser Thr Arg Leu 50 55 60 Ala Thr Gln Pro His Leu Val Gln Thr Gln Gln Val Gln Pro Gln Asn 65 70 75 80 Leu Gln Met Gln Gln Gln Asn Leu Gln Pro Ala Asn Ile Gln Gln Gln 85 90 95 Gln Ser Leu Gln Pro Pro Pro Pro Pro Pro Gln Pro His Leu Gly Val 100 105 110 Ser Ser Ala Ala Ser Gly His Leu Gly Arg Ser Phe Leu Ser Gly Glu 115 120 125 Pro Ser Gln Ala Asp Val Gln Pro Leu Gly Pro Ser Ser Leu Ala Val 130 135 140 His Thr Ile Leu Pro Gln Glu Ser Pro Ala Leu Pro Thr Ser Leu Pro 145 150 155 160 Ser Ser Leu Val Pro Pro Val Thr Ala Ala Gln Phe Leu Thr Pro Pro 165 170 175 Ser Gln His Ser Tyr Ser 180 60 66 PRT Homo sapiens 60 Tyr Ser Ser Pro Val Asp Asn Thr Pro Ser His Gln Leu Gln Val Pro 1 5 10 15 Glu His Pro Phe Leu Thr Pro Ser Pro Glu Ser Pro Asp Gln Trp Ser 20 25 30 Ser Ser Ser Pro His Ser Asn Val Ser Asp Trp Ser Glu Gly Val Ser 35 40 45 Ser Pro Pro Thr Ser Met Gln Ser Gln Ile Ala Arg Ile Pro Glu Ala 50 55 60 Phe Lys 65 61 57 PRT Danio rerio 61 Gly Ser Ile Val Tyr Leu Glu Ile Asp Asn Arg Leu Cys Ser Gln Gly 1 5 10 15 Ser Asp Asp Cys Phe Arg Asn Ala Asp Ser Ala Ala Glu Tyr Leu Gly 20 25 30 Ala Leu Ser Ala Arg Glu Met Leu Arg Phe Pro Tyr Pro Ile Lys Glu 35 40 45 Val Thr Ser Glu Lys Arg Glu Pro Ser 50 55 62 59 PRT Homo sapiens 62 Gly Ser Val Val Met Leu Glu Ile Asp Asn Arg Leu Cys Leu Gln Ser 1 5 10 15 Ala Glu Asn Asp His Cys Phe Pro Asp Ala Gln Ser Ala Ala Asp Tyr 20 25 30 Leu Gly Ala Leu Ser Ala Val Glu Arg Leu Asp Phe Pro Tyr Pro Leu 35 40 45 Arg Asp Val Arg Gly Glu Pro Leu Glu Ala Pro 50 55 63 57 PRT Mus musculus 63 Gly Ser Ile Val Tyr Leu Glu Ile Asp Asn Arg Gln Cys Val Gln Ser 1 5 10 15 Ser Ser Gln Cys Phe Gln Ser Ala Thr Asp Val Ala Ala Phe Leu Gly 20 25 30 Ala Leu Ala Ser Leu Gly Ser Leu Asn Ile Pro Tyr Lys Ile Glu Ala 35 40 45 Val Lys Ser Glu Pro Val Glu Pro Pro 50 55 64 57 PRT Rattus rattus 64 Gly Ser Ile Val Tyr Leu Glu Ile Asp Asn Arg Gln Cys Val Gln Ser 1 5 10 15 Ser Ser Gln Cys Phe Gln Ser Ala Thr Asp Val Ala Ala Phe Leu Gly 20 25 30 Ala Leu Ala Ser Leu Gly Ser Leu Asn Ile Pro Tyr Lys Ile Glu Ala 35 40 45 Val Lys Ser Glu Thr Val Glu Pro Pro 50 55 65 57 PRT Homo sapiens 65 Gly Ser Ile Val Tyr Leu Glu Ile Asp Asn Arg Gln Cys Val Gln Ala 1 5 10 15 Ser Ser Gln Cys Phe Gln Ser Ala Thr Asp Val Ala Ala Phe Leu Gly 20 25 30 Ala Leu Ala Ser Leu Gly Ser Leu Asn Ile Pro Tyr Lys Ile Glu Ala 35 40 45 Val Gln Ser Glu Thr Val Glu Pro Pro 50 55 66 57 PRT Gallus gallus 66 Gly Ser Ile Val Tyr Leu Glu Ile Asp Asn Arg Gln Cys Ile Gln Ser 1 5 10 15 Ser Ser Gln Cys Phe Gln Ser Ala Thr Asp Val Ala Ala Phe Leu Gly 20 25 30 Ala Leu Ala Ser Leu Gly Asn Leu Asn Ile Pro Tyr Lys Ile Glu Ala 35 40 45 Val Lys Ser Glu Thr Ala Glu Pro Ala 50 55 67 58 PRT Xenopus laevis 67 Gly Ser Ile Val Tyr Leu Glu Ile Asp Asn Arg Gln Cys Tyr Lys Ser 1 5 10 15 Ser Ser Gln Cys Phe Asn Ser Ala Thr Asp Val Ala Ala Phe Leu Gly 20 25 30 Ala Leu Ala Ser Leu Gly Ser Leu Asp Thr Leu Ser Tyr Lys Ile Glu 35 40 45 Ala Val Lys Ser Glu Asn Met Glu Thr Pro 50 55 68 55 PRT Homo sapiens 68 Gly Ser Lys Val Phe Leu Glu Ile Asp Asn Arg Gln Cys Val Gln Asp 1 5 10 15 Ser Asp His Cys Phe Lys Asn Thr Asp Ala Ala Ala Ala Leu Leu Ala 20 25 30 Ser His Ala Ile Gln Gly Thr Leu Ser Tyr Pro Leu Val Ser Val Val 35 40 45 Ser Glu Ser Leu Thr Pro Glu 50 55 69 60 PRT artificial sequence consensus sequence misc_feature (3)..(3) Xaa=Lys, Ile or Val misc_feature (5)..(5) Xaa=Phe, Tyr or Met misc_feature (12)..(12) Xaa=Gln or Leu misc_feature (14)..(14) Xaa=Val, Tyr, Ile, Leu or Ser misc_feature (15)..(15) Xaa=Gln or Lys misc_feature (18)..(18) Xaa=Asp, Ser, Ala, Glu or Gly misc_feature (19)..(19) Xaa=Ser or Asn misc_feature (20)..(20) Xaa=Asp or Ser misc_feature (21)..(21) Xaa=His, Gln or Asp misc_feature (24)..(24) Xaa=Lys, Asn, Pro, Gln or Arg misc_feature (25)..(25) Xaa=Asn, Ser or Asp misc_feature (26)..(26) Xaa=Thr or Ala misc_feature (27)..(27) Xaa=Asp, Thr or Gln misc_feature (28)..(28) Xaa=Ala, Asp or Ser misc_feature (29)..(29) Xaa=Ala or Val misc_feature (31)..(31) Xaa=Ala, Asp or Glu misc_feature (32)..(32) Xaa=Leu, Phe or Tyr misc_feature (34)..(34) Xaa=Ala or Gly misc_feature (35)..(35) Xaa=Ser or Ala misc_feature (36)..(36) Xaa=His or Leu misc_feature (37)..(37) Xaa=Ala or Ser misc_feature (38)..(38) Xaa=Ile, Ser or Ala misc_feature (39)..(39) Xaa=Gln, Leu, Val or Arg misc_feature (40)..(40) Xaa=Gly or Glu misc_feature (41)..(41) Xaa=Thr, Ser, Asn, Arg or Met misc_feature (43)..(43) Xaa=Ser, Asp, Gln or Arg misc_feature (45)..(45) Xaa=Leu, Ile or Phe misc_feature (46)..(46) Xaa=Ser or Pro misc_feature (48)..(48) Xaa=Pro or Lys misc_feature (49)..(49) Xaa=Leu or Ile misc_feature (50)..(50) Xaa=Val, Glu, Arg or Lys misc_feature (51)..(51) Xaa=Ser, Ala, Asp or Glu misc_feature (53)..(53) Xaa=Val, Lys, Gln, Arg or Thr misc_feature (54)..(54) Xaa=Ser or Gly misc_feature (56)..(56) Xaa=Ser, Asn, Thr, Pro or Lys misc_feature (57)..(57) Xaa=Leu, Met, Ala, Val or Arg misc_feature (58)..(58) Xaa=Thr or Glu misc_feature (59)..(59) Xaa=Pro, Thr or Ala misc_feature (60)..(60) X=Glu, Pro, Ala or Ser 69 Gly Ser Xaa Val Xaa Leu Glu Ile Asp Asn Arg Xaa Cys Xaa Xaa Ser 1 5 10 15 Ala Xaa Xaa Xaa Xaa Cys Phe Xaa Xaa Xaa Xaa Xaa Xaa Ala Xaa Xaa 20 25 30 Leu Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Leu Xaa Thr Xaa Xaa Tyr Xaa 35 40 45 Xaa Xaa Xaa Val Xaa Xaa Glu Xaa Xaa Xaa Xaa Xaa 50 55 60 70 58 PRT Homo sapiens 70 Gly Ser Ile Val Tyr Leu Glu Ile Asp Asn Arg Gln Cys Val Gln Ala 1 5 10 15 Ser Ser Gln Cys Phe Gln Ser Ala Thr Asp Val Ala Ala Phe Leu Gly 20 25 30 Ala Leu Ala Ser Leu Gly Ser Leu Asn Ile Pro Tyr Lys Ile Pro Glu 35 40 45 Ala Val Gln Ser Glu Thr Val Glu Pro Pro 50 55 71 57 PRT Homo sapiens 71 Gly Ser Ile Val Tyr Pro Glu Ile Asp Asn Arg Gln Cys Val Gln Ala 1 5 10 15 Ser Ser Gln Cys Phe Gln Ser Ala Thr Asp Val Ala Ala Phe Leu Gly 20 25 30 Ala Leu Ala Ser Leu Gly Ser Leu Asn Ile Pro Tyr Lys Ile Glu Ala 35 40 45 Val Gln Ser Glu Thr Val Glu Pro Pro 50 55 72 57 PRT Homo sapiens 72 Gly Ser Ile Asp Tyr Leu Glu Ile Asp Asn Arg Gln Cys Val Gln Ala 1 5 10 15 Ser Ser Gln Cys Phe Gln Ser Ala Thr Asp Val Ala Ala Phe Leu Gly 20 25 30 Ala Leu Ala Ser Leu Gly Ser Leu Asn Ile Pro Tyr Lys Ile Glu Ala 35 40 45 Val Gln Ser Glu Thr Val Glu Pro Pro 50 55 73 57 PRT Homo sapiens 73 Gly Ser Ile Val Tyr Leu Glu Asn Asp Asn Arg Gln Cys Val Gln Ala 1 5 10 15 Ser Ser Gln Cys Phe Gln Ser Ala Thr Asp Val Ala Ala Phe Leu Gly 20 25 30 Ala Leu Ala Ser Leu Gly Ser Leu Asn Ile Pro Tyr Lys Ile Glu Ala 35 40 45 Val Gln Ser Glu Thr Val Glu Pro Pro 50 55 74 57 PRT Homo sapiens 74 Gly Ser Ile Val Tyr Leu Glu Thr Asp Asn Arg Gln Cys Val Gln Ala 1 5 10 15 Ser Ser Gln Cys Phe Gln Ser Ala Thr Asp Val Ala Ala Phe Leu Gly 20 25 30 Ala Leu Ala Ser Leu Gly Ser Leu Asn Ile Pro Tyr Lys Ile Glu Ala 35 40 45 Val Gln Ser Glu Thr Val Glu Pro Pro 50 55 75 57 PRT Homo sapiens 75 Gly Ser Ile Val Tyr Leu Glu Ile Asp Asn Arg Gln Cys Val Gln Ala 1 5 10 15 Ser Ser Gln Cys Phe Gln Ser Ala Asn Asp Val Ala Ala Phe Leu Gly 20 25 30 Ala Leu Ala Ser Leu Gly Ser Leu Asn Ile Pro Tyr Lys Ile Glu Ala 35 40 45 Val Gln Ser Glu Thr Val Glu Pro Pro 50 55 76 7671 DNA Homo sapiens 76 atgccgccgc tcctggcgcc cctgctctgc ctggcgctgc tgcccgcgct cgccgcacga 60 ggcccgcgat gctcccagcc cggtgagacc tgcctgaatg gcgggaagtg tgaagcggcc 120 aatggcacgg aggcctgcgt ctgtggcggg gccttcgtgg gcccgcgatg ccaggacccc 180 aacccgtgcc tcagcacccc ctgcaagaac gccgggacat gccacgtggt ggaccgcaga 240 ggcgtggcag actatgcctg cagctgtgcc ctgggcttct ctgggcccct ctgcctgaca 300 cccctggaca atgcctgcct caccaacccc tgccgcaacg ggggcacctg cgacctgctc 360 acgctgacgg agtacaagtg ccgctgcccg cccggctggt cagggaaatc gtgccagcag 420 gctgacccgt gcgcctccaa cccctgcgcc aacggtggcc agtgcctgcc cttcgaggcc 480 tcctacatct gccactgccc acccagcttc catggcccca cctgccggca ggatgtcaac 540 gagtgtggcc agaagcccgg gctttgccgc cacggaggca cctgccacaa cgaggtcggc 600 tcctaccgct gcgtctgccg cgccacccac actggcccca actgcgagcg gccctacgtg 660 ccctgcagcc cctcgccctg ccagaacggg ggcacctgcc gccccacggg cgacgtcacc 720 cacgagtgtg cctgcctgcc aggcttcacc ggccagaact gtgaggaaaa tatcgacgat 780 tgtccaggaa acaactgcaa gaacgggggt gcctgtgtgg acggcgtgaa cacctacaac 840 tgccgctgcc cgccagagtg gacaggtcag tactgtaccg aggatgtgga cgagtgccag 900 ctgatgccaa atgcctgcca gaacggcggg acctgccaca acacccacgg tggctacaac 960 tgcgtgtgtg tcaacggctg gactggtgag gactgcagcg agaacattga tgactgtgcc 1020 agcgccgcct gcttccacgg cgccacctgc catgaccgtg tggcctcctt ctactgcgag 1080 tgtccccatg gccgcacagg tctgctgtgc cacctcaacg acgcatgcat cagcaacccc 1140 tgtaacgagg gctccaactg cgacaccaac cctgtcaatg gcaaggccat ctgcacctgc 1200 ccctcggggt acacgggccc ggcctgcagc caggacgtgg atgagtgctc gctgggtgcc 1260 aacccctgcg agcatgcggg caagtgcatc aacacgctgg gctccttcga gtgccagtgt 1320 ctgcagggct acacgggccc ccgatgcgag atcgacgtca acgagtgcgt ctcgaacccg 1380 tgccagaacg acgccacctg cctggaccag attggggagt tccagtgcat ctgcatgccc 1440 ggctacgagg gtgtgcactg cgaggtcaac acagacgagt gtgccagcag cccctgcctg 1500 cacaatggcc gctgcctgga caagatcaat gagttccagt gcgagtgccc cacgggcttc 1560 actgggcatc tgtgccagta cgatgtggac gagtgtgcca gcaccccctg caagaatggt 1620 gccaagtgcc tggacggacc caacacttac acctgtgtgt gcacggaagg gtacacgggg 1680 acgcactgcg aggtggacat cgatgagtgc gaccccgacc cctgccacta cggctcctgc 1740 aaggacggcg tcgccacctt cacctgcctc tgccgcccag gctacacggg ccaccactgc 1800 gagaccaaca tcaacgagtg ctccagccag ccctgccgcc acgggggcac ctgccaggac 1860 cgcgacaacg cctacctctg cttctgcctg aaggggacca caggacccaa ctgcgagatc 1920 aacctggatg actgtgccag cagcccctgc gactcgggca cctgtctgga caagatcgat 1980 ggctacgagt gtgcctgtga gccgggctac acagggagca tgtgtaacat caacatcgat 2040 gagtgtgcgg gcaacccctg ccacaacggg

ggcacctgcg aggacggcat caatggcttc 2100 acctgccgct gccccgaggg ctaccacgac cccacctgcc tgtctgaggt caatgagtgc 2160 aacagcaacc cctgcgtcca cggggcctgc cgggacagcc tcaacgggta caagtgcgac 2220 tgtgaccctg ggtggagtgg gaccaactgt gacatcaaca acaatgagtg tgaatccaac 2280 ccttgtgtca acggcggcac ctgcaaagac atgaccagtg gctacgtgtg cacctgccgg 2340 gagggcttca gcggtcccaa ctgccagacc aacatcaacg agtgtgcgtc caacccatgt 2400 ctgaaccagg gcacgtgtat tgacgacgtt gccgggtaca agtgcaactg cctgctgccc 2460 tacacaggtg ccacgtgtga ggtggtgctg gccccgtgtg cccccagccc ctgcagaaac 2520 ggcggggagt gcaggcaatc cgaggactat gagagcttct cctgtgtctg ccccacgggc 2580 tggcaagcag ggcagacctg tgaggtcgac atcaacgagt gcgttctgag cccgtgccgg 2640 cacggcgcat cctgccagaa cacccacggc ggctaccgct gccactgcca ggccggctac 2700 agtgggcgca actgcgagac cgacatcgac gactgccggc ccaacccgtg tcacaacggg 2760 ggctcctgca cagacggcat caacacggcc ttctgcgact gcctgcccgg cttccggggc 2820 actttctgtg aggaggacat caacgagtgt gccagtgacc cctgccgcaa cggggccaac 2880 tgcacggact gcgtggacag ctacacgtgc acctgccccg caggcttcag cgggatccac 2940 tgtgagaaca acacgcctga ctgcacagag agctcctgct tcaacggtgg cacctgcgtg 3000 gacggcatca actcgttcac ctgcctgtgt ccacccggct tcacgggcag ctactgccag 3060 cacgatgtca atgagtgcga ctcacagccc tgcctgcatg gcggcacctg tcaggacggc 3120 tgcggctcct acaggtgcac ctgcccccag ggctacactg gccccaactg ccagaacctt 3180 gtgcactggt gtgactcctc gccctgcaag aacggcggca aatgctggca gacccacacc 3240 cagtaccgct gcgagtgccc cagcggctgg accggccttt actgcgacgt gcccagcgtg 3300 tcctgtgagg tggctgcgca gcgacaaggt gttgacgttg cccgcctgtg ccagcatgga 3360 gggctctgtg tggacgcggg caacacgcac cactgccgct gccaggcggg ctacacaggc 3420 agctactgtg aggacctggt ggacgagtgc tcacccagcc cctgccagaa cggggccacc 3480 tgcacggact acctgggcgg ctactcctgc aagtgcgtgg ccggctacca cggggtgaac 3540 tgctctgagg agatcgacga gtgcctctcc cacccctgcc agaacggggg cacctgcctc 3600 gacctcccca acacctacaa gtgctcctgc ccacggggca ctcagggtgt gcactgtgag 3660 atcaacgtgg acgactgcaa tccccccgtt gaccccgtgt cccggagccc caagtgcttt 3720 aacaacggca cctgcgtgga ccaggtgggc ggctacagct gcacctgccc gccgggcttc 3780 gtgggtgagc gctgtgaggg ggatgtcaac gagtgcctgt ccaatccctg cgacgcccgt 3840 ggcacccaga actgcgtgca gcgcgtcaat gacttccact gcgagtgccg tgctggtcac 3900 accgggcgcc gctgcgagtc cgtcatcaat ggctgcaaag gcaagccctg caagaatggg 3960 ggcacctgcg ccgtggcctc caacaccgcc cgcgggttca tctgcaagtg ccctgcgggc 4020 ttcgagggcg ccacgtgtga gaatgacgct cgtacctgcg gcagcctgcg ctgcctcaac 4080 ggcggcacat gcatctccgg cccgcgcagc cccacctgcc tgtgcctggg ccccttcacg 4140 ggccccgaat gccagttccc ggccagcagc ccctgcctgg gcggcaaccc ctgctacaac 4200 caggggacct gtgagcccac atccgagagc cccttctacc gttgcctgtg ccccgccaaa 4260 ttcaacgggc tcttgtgcca catcctggac tacagcttcg ggggtggggc cgggcgcgac 4320 atccccccgc cgctgatcga ggaggcgtgc gagctgcccg agtgccagga ggacgcgggc 4380 aacaaggtct gcagcctgca gtgcaacaac cacgcgtgcg gctgggacgg cggtgactgc 4440 tccctcaact tcaatgaccc ctggaagaac tgcacgcagt ctctgcagtg ctggaagtac 4500 ttcagtgacg gccactgtga cagccagtgc aactcagccg gctgcctctt cgacggcttt 4560 gactgccagc gtgcggaagg ccagtgcaac cccctgtacg accagtactg caaggaccac 4620 ttcagcgacg ggcactgcga ccagggctgc aacagcgcgg agtgcgagtg ggacgggctg 4680 gactgtgcgg agcatgtacc cgagaggctg gcggccggca cgctggtggt ggtggtgctg 4740 atgccgccgg agcagctgcg caacagctcc ttccacttcc tgcgggagct cagccgcgtg 4800 ctgcacacca acgtggtctt caagcgtgac gcacacggcc agcagatgat cttcccctac 4860 tacggccgcg aggaggagct gcgcaagcac cccatcaagc gtgccgccga gggctgggcc 4920 gcacctgacg ccctgctggg ccaggtgaag gcctcgctgc tccctggtgg cagcgagggt 4980 gggcggcggc ggagggagct ggaccccatg gacgtccgcg gctccatcgt ctacctggag 5040 attgacaacc ggcagtgtgt gcaggcctcc tcgcagtgct tccagagtgc caccgacgtg 5100 gccgcattcc tgggagcgct cgcctcgctg ggcagcctca acatccccta caagatcgag 5160 gccgtgcaga gtgagaccgt ggagccgccc ccgccggcgc agctgcactt catgtacgtg 5220 gcggcggccg cctttgtgct tctgttcttc gtgggctgcg gggtgctgct gtcccgcaag 5280 cgccggcggc agcatggcca gctctggttc cctgagggct tcaaagtgtc tgaggccagc 5340 aagaagaagc ggcgggagcc cctcggcgag gactccgtgg gcctcaagcc cctgaagaac 5400 gcttcagacg gtgccctcat ggacgacaac cagaatgagt ggggggacga ggacctggag 5460 accaagaagt tccggttcga ggagcccgtg gttctgcctg acctggacga ccagacagac 5520 caccggcagt ggactcagca gcacctggat gccgctgacc tgcgcatgtc tgccatggcc 5580 cccacaccgc cccagggtga ggttgacgcc gactgcatgg acgtcaatgt ccgcgggcct 5640 gatggcttca ccccgctcat gatcgcctcc tgcagcgggg gcggcctgga gacgggcaac 5700 agcgaggaag aggaggacgc gccggccgtc atctccgact tcatctacca gggcgccagc 5760 ctgcacaacc agacagaccg cacgggcgag accgccttgc acctggccgc ccgctactca 5820 cgctctgatg ccgccaagcg cctgctggag gccagcgcag atgccaacat ccaggacaac 5880 atgggccgca ccccgctgca tgcggctgtg tctgccgacg cacaaggtgt cttccagatc 5940 ctgatccgga accgagccac agacctggat gcccgcatgc atgatggcac gacgccactg 6000 atcctggctg cccgcctggc cgtggagggc atgctggagg acctcatcaa ctcacacgcc 6060 gacgtcaacg ccgtagatga cctgggcaag tccgccctgc actgggccgc cgccgtgaac 6120 aatgtggatg ccgcagttgt gctcctgaag aacggggcta acaaagatat gcagaacaac 6180 agggaggaga cacccctgtt tctggccgcc cgggagggca gctacgagac cgccaaggtg 6240 ctgctggacc actttgccaa ccgggacatc acggatcata tggaccgcct gccgcgcgac 6300 atcgcacagg agcgcatgca tcacgacatc gtgaggctgc tggacgagta caacctggtg 6360 cgcagcccgc agctgcacgg agccccgctg gggggcacgc ccaccctgtc gcccccgctc 6420 tgctcgccca acggctacct gggcagcctc aagcccggcg tgcagggcaa gaaggtccgc 6480 aagcccagca gcaaaggcct ggcctgtgga agcaaggagg ccaaggacct caaggcacgg 6540 aggaagaagt cccaggacgg caagggctgc ctgctggaca gctccggcat gctctcgccc 6600 gtggactccc tggagtcacc ccatggctac ctgtcagacg tggcctcgcc gccactgctg 6660 ccctccccgt tccagcagtc tccgtccgtg cccctcaacc acctgcctgg gatgcccgac 6720 acccacctgg gcatcgggca cctgaacgtg gcggccaagc ccgagatggc ggcgctgggt 6780 gggggcggcc ggctggcctt tgagactggc ccacctcgtc tctcccacct gcctgtggcc 6840 tctggcacca gcaccgtcct gggctccagc agcggagggg ccctgaattt cactgtgggc 6900 gggtccacca gtttgaatgg tcaatgcgag tggctgtccc ggctgcagag cggcatggtg 6960 ccgaaccaat acaaccctct gcgggggagt gtggcaccag gccccctgag cacacaggcc 7020 ccctccctgc agcatggcat ggtaggcccg ctgcacagta gccttgctgc cagcgccctg 7080 tcccagatga tgagctacca gggcctgccc agcacccggc tggccaccca gcctcacctg 7140 gtgcagaccc agcaggtgca gccacaaaac ttacagatgc agcagcagaa cctgcagcca 7200 gcaaacatcc agcagcagca aagcctgcag ccgccaccac caccaccaca gccgcacctt 7260 ggcgtgagct cagcagccag cggccacctg ggccggagct tcctgagtgg agagccgagc 7320 caggcagacg tgcagccact gggccccagc agcctggcgg tgcacactat tctgccccag 7380 gagagccccg ccctgcccac gtcgctgcca tcctcgctgg tcccacccgt gaccgcagcc 7440 cagttcctga cgcccccctc gcagcacagc tactcctcgc ctgtggacaa cacccccagc 7500 caccagctac aggtgcctga gcaccccttc ctcaccccgt cccctgagtc ccctgaccag 7560 tggtccagct cgtccccgca ttccaacgtc tccgactggt ccgagggcgt ctccagccct 7620 cccaccagca tgcagtccca gatcgcccgc attccggagg ccttcaagta a 7671 77 52 PRT Homo sapiens 77 Ala Ala Gly Thr Leu Val Val Val Val Leu Met Pro Pro Glu Gln Leu 1 5 10 15 Arg Asn Ser Ser Phe His Phe Leu Arg Glu Leu Ser Arg Val Leu His 20 25 30 Thr Asn Val Val Phe Lys Arg Asp Ala His Gly Gln Gln Met Ile Phe 35 40 45 Pro Tyr Tyr Gly 50 78 38 DNA Homo sapiens 78 tttgaattcg ggctggactg tgcggagcat gtacccga 38 79 37 DNA Homo sapiens 79 tttggatcct ccggaatgcg ggcgatctgg gactgca 37 80 20 DNA Homo sapiens 80 agccccctgt acgaccagta 20 81 18 DNA Homo sapiens 81 cttgcgcagc tcctcctc 18 82 20 DNA Homo sapiens 82 gaccagtact gcaaggacca 20 83 18 DNA Homo sapiens 83 tcctcgcggc cgtagtag 18 84 18 DNA Homo sapiens 84 gtgctgcaca ccaacgtg 18 85 18 DNA Homo sapiens 85 gagggcccag gagagttg 18 86 20 DNA Homo sapiens 86 gcacacggcc agcagatgat 20 87 17 DNA Homo sapiens 87 cgccgggtct cactcac 17 88 18 DNA Homo sapiens 88 gtggcgtcat gggcctca 18 89 20 DNA Homo sapiens 89 tagcaactgg cacaaacagc 20 90 18 DNA Homo sapiens 90 catgggcctc agtgtcct 18 91 20 DNA Homo sapiens 91 gcacaaacag ccagcgtgtc 20 92 18 DNA Homo sapiens 92 gcagcatggc atggtagg 18 93 25 DNA Homo sapiens 93 aacatgtgtt ttaaaaaggc tcctc 25 94 20 DNA Homo sapiens 94 aaacatccag cagcagcaaa 20 95 21 DNA Homo sapiens 95 cacaggcgag gagtagctgt g 21 96 18 DNA Homo sapiens 96 gtgaccgcag cccagttc 18 97 18 DNA Homo sapiens 97 aaaggaagcc ggggtctc 18 98 21 DNA Homo sapiens 98 agactggccc acctcgtctc t 21 99 20 DNA Homo sapiens 99 gctctccact caggaagctc 20 100 20 DNA Homo sapiens 100 cgtctctccc acctgcctgt 20 101 18 DNA Homo sapiens 101 ctgagctcac gccaaggt 18 102 48 PRT Homo sapiens 102 Ala Ala Gly Thr Leu Val Glu Val Val Leu Met Pro Pro Glu Gln Leu 1 5 10 15 Arg Asn Ser Ser Phe His Phe Leu Arg Glu Leu Ser Arg Val Leu His 20 25 30 Thr Asn Val Val Phe Lys Arg Asp Ala His Gly Gln Gln Met Ile Phe 35 40 45 103 48 PRT Homo sapiens 103 Ala Ala Gly Thr Leu Val Val Glu Val Leu Met Pro Pro Glu Gln Leu 1 5 10 15 Arg Asn Ser Ser Phe His Phe Leu Arg Glu Leu Ser Arg Val Leu His 20 25 30 Thr Asn Val Val Phe Lys Arg Asp Ala His Gly Gln Gln Met Ile Phe 35 40 45 104 48 PRT Homo sapiens 104 Ala Ala Gly Thr Leu Val Val Val Gly Leu Met Pro Pro Glu Gln Leu 1 5 10 15 Arg Asn Ser Ser Phe His Phe Leu Arg Glu Leu Ser Arg Val Leu His 20 25 30 Thr Asn Val Val Phe Lys Arg Asp Ala His Gly Gln Gln Met Ile Phe 35 40 45 105 48 PRT Homo sapiens 105 Ala Ala Gly Thr Leu Val Val Val Val Leu Met Pro Pro Glu Gln Pro 1 5 10 15 Arg Asn Ser Ser Phe His Phe Leu Arg Glu Leu Ser Arg Val Leu His 20 25 30 Thr Asn Val Val Phe Lys Arg Asp Ala His Gly Gln Gln Met Ile Phe 35 40 45 106 48 PRT Homo sapiens 106 Ala Ala Gly Thr Leu Val Val Val Val Leu Met Pro Pro Glu Gln Gln 1 5 10 15 Arg Asn Ser Ser Phe His Phe Leu Arg Glu Leu Ser Arg Val Leu His 20 25 30 Thr Asn Val Val Phe Lys Arg Asp Ala His Gly Gln Gln Met Ile Phe 35 40 45 107 48 PRT Homo sapiens 107 Ala Ala Gly Thr Leu Val Val Val Val Leu Met Pro Pro Val Gln Leu 1 5 10 15 Arg Asn Ser Ser Phe His Phe Leu Arg Glu Leu Ser Arg Val Leu His 20 25 30 Thr Asn Val Val Phe Lys Arg Asp Ala His Gly Gln Gln Met Ile Phe 35 40 45 108 48 PRT Homo sapiens 108 Ala Ala Gly Thr Leu Val Val Val Val Leu Met Pro Pro Glu Gln Leu 1 5 10 15 Arg Asn Ser Ser Phe His Phe Leu Arg Glu Pro Ser Arg Val Leu His 20 25 30 Thr Asn Val Val Phe Lys Arg Asp Ala His Gly Gln Gln Met Ile Phe 35 40 45 109 48 PRT Homo sapiens 109 Ala Ala Gly Thr Leu Val Val Val Val Leu Met Pro Pro Glu Gln Leu 1 5 10 15 Arg Asn Ser Ser Phe His Phe Leu Arg Glu His Ser Arg Val Leu His 20 25 30 Thr Asn Val Val Phe Lys Arg Asp Ala His Gly Gln Gln Met Ile Phe 35 40 45 110 48 PRT Homo sapiens 110 Ala Ala Gly Thr Leu Val Val Val Val Leu Met Pro Pro Glu Gln Leu 1 5 10 15 Arg Asn Ser Ser Phe His Phe Leu Arg Glu Leu Ile Arg Val Leu His 20 25 30 Thr Asn Val Val Phe Lys Arg Asp Ala His Gly Gln Gln Met Ile Phe 35 40 45 111 48 PRT Homo sapiens 111 Ala Ala Gly Thr Leu Val Val Val Val Leu Met Pro Pro Glu Gln Leu 1 5 10 15 Arg Asn Ser Ser Phe His Phe Leu Arg Glu Leu Ser Arg Val Leu His 20 25 30 Asn Asn Val Val Phe Lys Arg Asp Ala His Gly Gln Gln Met Ile Phe 35 40 45 112 48 PRT Homo sapiens 112 Ala Ala Gly Thr Leu Val Val Val Val Leu Met Pro Pro Glu Gln Leu 1 5 10 15 Arg Asn Ser Ser Phe His Phe Leu Arg Glu Leu Ser Arg Val Leu His 20 25 30 Thr Asn Glu Val Phe Lys Arg Asp Ala His Gly Gln Gln Met Ile Phe 35 40 45 113 48 PRT Homo sapiens 113 Ala Ala Gly Thr Leu Val Val Val Val Leu Met Pro Pro Glu Gln Leu 1 5 10 15 Arg Asn Ser Ser Phe His Phe Leu Arg Glu Leu Ser Arg Val Leu His 20 25 30 Thr Asn Gly Val Phe Lys Arg Asp Ala His Gly Gln Gln Met Ile Phe 35 40 45 114 48 PRT Homo sapiens 114 Ala Ala Gly Thr Leu Val Val Val Val Leu Met Pro Pro Glu Gln Leu 1 5 10 15 Arg Asn Ser Ser Phe His Phe Leu Arg Glu Leu Ser Arg Val Leu His 20 25 30 Thr Asn Val Val Phe Lys Ser Asp Ala His Gly Gln Gln Met Ile Phe 35 40 45 115 48 PRT Homo sapiens 115 Ala Ala Gly Thr Leu Val Val Val Val Leu Met Pro Pro Glu Gln Leu 1 5 10 15 Arg Asn Ser Ser Phe His Phe Leu Arg Glu Leu Ser Arg Val Leu His 20 25 30 Thr Asn Val Val Phe Lys Arg Asp Ala His Gly Gln Gln Met Asn Phe 35 40 45 116 48 PRT Homo sapiens 116 Ala Ala Gly Thr Leu Val Val Val Val Leu Met Pro Pro Glu Gln Leu 1 5 10 15 Arg Asn Ser Ser Phe His Phe Leu Arg Glu Leu Ser Arg Val Leu His 20 25 30 Thr Asn Val Val Phe Lys Arg Asp Ala His Gly Gln Gln Met Thr Phe 35 40 45 117 48 PRT Homo sapiens 117 Ala Ala Gly Thr Leu Val Val Val Val Leu Met Pro Pro Glu Gln Leu 1 5 10 15 Arg Asn Ser Ser Phe Arg Ser Leu Arg Glu Leu Ser Arg Val Leu His 20 25 30 Thr Asn Val Val Phe Lys Arg Asp Ala His Gly Gln Gln Met Ile Phe 35 40 45 118 47 PRT Homo sapiens 118 Ala Ala Gly Thr Leu Val Val Val Val Leu Met Pro Glu Gln Leu Arg 1 5 10 15 Asn Ser Ser Phe His Phe Leu Arg Glu Leu Ser Arg Val Leu His Thr 20 25 30 Asn Val Val Phe Lys Arg Asp Ala His Gly Gln Gln Met Ile Phe 35 40 45 119 53 PRT Homo sapiens 119 Ala Ala Gly Thr Leu Val Val Val Val Leu Met Pro Pro Glu Gln Leu 1 5 10 15 Arg Asn Ser Ser Phe His Phe Ala Leu Arg Glu Leu Ser Arg Val Leu 20 25 30 His Thr Asn Val Val Phe Lys Arg Asp Ala His Gly Gln Gln Met Ile 35 40 45 Phe Pro Tyr Tyr Gly 50 120 53 PRT Homo sapiens 120 Ala Ala Gly Thr Leu Val Val Val Val Leu Met Pro Pro Glu Gln Leu 1 5 10 15 Arg Asn Ser Ser Phe His Leu Tyr Leu Arg Glu Leu Ser Arg Val Leu 20 25 30 His Thr Asn Val Val Phe Lys Arg Asp Ala His Gly Gln Gln Met Ile 35 40 45 Phe Pro Tyr Tyr Gly 50 121 54 PRT Homo sapiens 121 Ala Ala Gly Thr Leu Val Val Val Val Leu Met Pro Pro Glu Gln Leu 1 5 10 15 Arg Asn Ser Ser Phe His Ser His Leu Lys Arg Glu Leu Ser Arg Val 20 25 30 Leu His Thr Asn Val Val Phe Lys Arg Asp Ala His Gly Gln Gln Met 35 40 45 Ile Phe Pro Tyr Tyr Gly 50 122 56 PRT Homo sapiens 122 Ala Ala Gly Thr Leu Val Val Val Val Leu Met Pro Pro Glu Gln Leu 1 5 10 15 Arg Asn Ser Ser Phe His Phe Leu Arg Glu Leu Ser Arg Val Leu His 20 25 30 Thr Asn Val Leu Pro Pro Pro Ala Phe Lys Arg Asp Ala His Gly Gln 35 40 45 Gln Met Ile Phe Pro Tyr Tyr Gly 50 55 123 54 PRT Homo sapiens 123 Ala Ala Gly Thr Leu Val Val Val Val Leu Met Pro Pro Glu Gln Leu 1 5 10 15 Arg Asn Ser Ser Phe His Phe Leu Arg Glu Leu Ser Arg Val Leu His 20 25 30 Thr Asn Val Leu Gly Arg Phe Lys Arg Asp Ala His Gly Gln Gln Met 35 40 45 Ile Phe Pro Tyr Tyr Gly 50

124 57 PRT Homo sapiens 124 Ala Ala Gly Thr Leu Val Val Val Val Leu Met Pro Pro Glu Gln Leu 1 5 10 15 Arg Asn Ser Ser Phe His Phe Leu Arg Glu Leu Ser Arg Val Leu His 20 25 30 Thr Asn Val Val Phe Glu Arg Ala Arg Ser Lys Arg Asp Ala His Gly 35 40 45 Gln Gln Met Ile Phe Pro Tyr Tyr Gly 50 55 125 53 PRT Homo sapiens 125 Ala Ala Gly Thr Leu Val Val Val Val Leu Met Pro Pro Glu Gln Leu 1 5 10 15 Arg Asn Ser Ser Phe His Phe Leu Arg Glu Leu Ser Arg Val Leu His 20 25 30 Thr Asn Val Val Phe His Lys Arg Asp Ala His Gly Gln Gln Met Ile 35 40 45 Phe Pro Tyr Tyr Gly 50 126 53 PRT Homo sapiens 126 Ala Ala Gly Thr Leu Val Val Val Val Leu Met Pro Pro Glu Gln Leu 1 5 10 15 Arg Asn Ser Ser Phe His Phe Leu Arg Glu Leu Ser Arg Val Leu His 20 25 30 Thr Asn Val Val Phe Asp Lys Arg Asp Ala His Gly Gln Gln Met Ile 35 40 45 Phe Pro Tyr Tyr Gly 50 127 53 PRT Homo sapiens 127 Ala Ala Gly Thr Leu Val Val Val Val Leu Met Pro Pro Glu Gln Leu 1 5 10 15 Arg Asn Ser Ser Phe His Phe Leu Arg Glu Leu Ser Arg Val Leu His 20 25 30 Thr Asn Val Val Phe Lys Arg Asp Ala His Gly Gln Gln Met Ile Leu 35 40 45 Gly Pro Tyr Tyr Gly 50 128 57 PRT Homo sapiens 128 Gly Ser Ile Val Tyr Leu Glu Ile Asp Asn Arg Gln Cys Val Gln Ala 1 5 10 15 Ser Ser Gln Cys Phe Gln Ser Ala Thr Asp Val Ala Ala Phe Leu Gly 20 25 30 Ala Leu Ala Ser Leu Gly Ser Leu Asn Ile Pro Tyr Lys Thr Glu Ala 35 40 45 Val Gln Ser Glu Thr Val Glu Pro Pro 50 55 129 57 PRT Homo sapiens 129 Gly Ser Ile Val Tyr Leu Glu Ile Asp Asn Arg Gln Cys Val Gln Ala 1 5 10 15 Ser Ser Gln Cys Phe Gln Ser Ala Thr Asp Val Ala Pro Phe Leu Gly 20 25 30 Ala Leu Ala Ser Leu Gly Ser Leu Asn Ile Pro Tyr Lys Ile Glu Ala 35 40 45 Val Gln Ser Glu Thr Val Glu Pro Pro 50 55 130 57 PRT Homo sapiens 130 Gly Ser Ile Val Tyr Leu Glu Ile Asp Asn Arg Gln Cys Val Gln Ala 1 5 10 15 Ser Ser Gln Cys Phe Gln Ser Ala Thr Asp Val Ala Ala Phe Leu Gly 20 25 30 Ala Leu Ala Ser Pro Gly Ser Leu Asn Ile Pro Tyr Lys Ile Glu Ala 35 40 45 Val Gln Ser Glu Thr Val Glu Pro Pro 50 55 131 57 PRT Homo sapiens 131 Gly Ser Ile Val Tyr Leu Glu Ile Asp Asn Arg Gln Cys Val Gln Ala 1 5 10 15 Ser Ser Gln Cys Phe Gln Leu Ala Thr Asp Val Ala Ala Phe Leu Gly 20 25 30 Ala Leu Ala Ser Leu Gly Ser Leu Asn Ile Pro Tyr Lys Ile Glu Ala 35 40 45 Val Gln Ser Glu Thr Val Glu Pro Pro 50 55 132 60 PRT Homo sapiens 132 Gly Ser Ile Val Ser Thr Leu Tyr Leu Glu Ile Asp Asn Arg Gln Cys 1 5 10 15 Val Gln Ala Ser Ser Gln Cys Phe Gln Ser Ala Thr Asp Val Ala Ala 20 25 30 Phe Leu Gly Ala Leu Ala Ser Leu Gly Ser Leu Asn Ile Pro Tyr Lys 35 40 45 Ile Glu Ala Val Gln Ser Glu Thr Val Glu Pro Pro 50 55 60 133 57 PRT Homo sapiens 133 Gly Ser Ile Val Tyr Leu Glu Ile Asp Asn Arg Gln Cys Val Gln Ala 1 5 10 15 Ser Ser Gln Cys Phe Gln Ser Ala Asn Asp Val Ala Ala Phe Leu Gly 20 25 30 Ala Leu Ala Ser Leu Gly Ser Leu Asn Ile Pro Tyr Lys Ile Glu Ala 35 40 45 Val Gln Ser Glu Thr Val Glu Pro Pro 50 55 134 57 PRT Homo sapiens 134 Gly Ser Ile Val Tyr Leu Glu Asn Asp Asn Arg Gln Cys Val Gln Ala 1 5 10 15 Ser Ser Gln Cys Phe Gln Ser Ala Thr Asp Val Ala Ala Phe Leu Gly 20 25 30 Ala Leu Ala Ser Leu Gly Ser Leu Asn Ile Pro Tyr Lys Ile Glu Ala 35 40 45 Val Gln Ser Glu Thr Val Glu Pro Pro 50 55 135 57 PRT Homo sapiens 135 Gly Ser Ile Asp Tyr Leu Glu Ile Asp Asn Arg Gln Cys Val Gln Ala 1 5 10 15 Ser Ser Gln Cys Phe Gln Ser Ala Thr Asp Val Ala Ala Phe Leu Gly 20 25 30 Ala Leu Ala Ser Leu Gly Ser Leu Asn Ile Pro Tyr Lys Ile Glu Ala 35 40 45 Val Gln Ser Glu Thr Val Glu Pro Pro 50 55 136 57 PRT Homo sapiens 136 Gly Ser Ile Val Tyr Pro Glu Ile Asp Asn Arg Gln Cys Val Gln Ala 1 5 10 15 Ser Ser Gln Cys Phe Gln Ser Ala Thr Asp Val Ala Ala Phe Leu Gly 20 25 30 Ala Leu Ala Ser Leu Gly Ser Leu Asn Ile Pro Tyr Lys Ile Glu Ala 35 40 45 Val Gln Ser Glu Thr Val Glu Pro Pro 50 55 137 70 PRT Homo sapiens 137 Gly Ser Ile Val Tyr Leu Glu Ile Asp Asn Arg Gln Cys Val Gln Ala 1 5 10 15 Ser Ser Gln Cys Phe Gln Ser Ala Thr Asp Val Ala Ala Phe Leu Gly 20 25 30 Ala Leu Ala Ser Leu Gly Ser Leu Asn Ile Pro Tyr Lys Ile Glu Arg 35 40 45 Leu Gly Ser Leu Asn Ile Pro Tyr Lys Ile Glu Ala Val Val Gln Ser 50 55 60 Glu Thr Val Glu Pro Pro 65 70 138 81 PRT Homo sapiens 138 Trp Leu Ser Arg Leu Gln Ser Gly Met Val Pro Asn Gln Tyr Asn Pro 1 5 10 15 Leu Arg Gly Ser Val Ala Pro Gly Pro Leu Ser Thr Gln Ala Pro Ser 20 25 30 Leu Gln His Gly Met Val Gly Pro Leu His Ser Ser Leu Ala Ala Ser 35 40 45 Ala Leu Ser Gln Met Met Ser Tyr Gln Gly Leu Pro Ser Thr Arg Leu 50 55 60 Ala Thr Gln Pro His Leu Val Gln Thr Gln Gln Val Gln Pro Gln Asn 65 70 75 80 Leu 139 168 PRT Homo sapiens 139 Trp Leu Ser Arg Leu Gln Ser Gly Met Val Pro Asn Gln Tyr Asn Pro 1 5 10 15 Leu Arg Gly Ser Val Ala Pro Gly Pro Leu Ser Thr Gln Ala Pro Ser 20 25 30 Leu Gln His Gly Met Val Gly Pro Leu His Ser Ser Leu Ala Ala Ser 35 40 45 Ala Leu Ser Gln Met Met Ser Tyr Gln Gly Leu Pro Ser Thr Arg Leu 50 55 60 Ala Thr Gln Pro His Leu Val Gln Thr Gln Gln Val Gln Pro Gln Asn 65 70 75 80 Leu Gln Met Gln Gln Gln Asn Leu Gln Pro Ala Asn Ile Gln Gln Gln 85 90 95 Gln Ser Leu Gln Pro Pro Pro Pro Pro Pro Gln Pro His Leu Gly Val 100 105 110 Ser Ser Ala Ala Ser Gly His Leu Gly Arg Ser Phe Leu Ser Gly Glu 115 120 125 Pro Ser Gln Ala Asp Val Gln Pro Leu Gly Pro Ser Ser Leu Ala Val 130 135 140 His Thr Ile Leu Pro Gln Glu Ser Pro Ala Leu Pro Thr Ser Leu Pro 145 150 155 160 Ser Leu Ile Leu Trp Ser His Pro 165 140 134 PRT Homo sapiens 140 Trp Leu Ser Arg Leu Gln Ser Gly Met Val Pro Asn Gln Tyr Asn Pro 1 5 10 15 Leu Arg Gly Ser Val Ala Pro Gly Pro Leu Ser Thr Gln Ala Pro Ser 20 25 30 Leu Gln His Gly Met Val Gly Pro Leu His Ser Ser Leu Ala Ala Ser 35 40 45 Ala Leu Ser Gln Met Met Ser Tyr Gln Gly Leu Pro Ser Thr Arg Leu 50 55 60 Ala Thr Gln Pro His Leu Val Gln Thr Gln Gln Val Gln Pro Gln Asn 65 70 75 80 Leu Gln Met Gln Gln Gln Asn Leu Gln Pro Ala Asn Ile Gln Gln Gln 85 90 95 Gln Ser Leu Gln Pro Pro Pro Pro Pro Pro Gln Pro His Leu Gly Val 100 105 110 Ser Ser Ala Ala Ser Gly His Leu Gly Arg Ser Phe Leu Ser Gly Glu 115 120 125 Pro Ser Gln Ala Val Leu 130 141 116 PRT Homo sapiens 141 Trp Leu Ser Arg Leu Gln Ser Gly Met Val Pro Asn Gln Tyr Asn Pro 1 5 10 15 Leu Arg Gly Ser Val Ala Pro Gly Pro Leu Ser Thr Gln Ala Pro Ser 20 25 30 Leu Gln His Gly Met Val Gly Pro Leu His Ser Ser Leu Ala Ala Ser 35 40 45 Ala Leu Ser Gln Met Met Ser Tyr Gln Gly Leu Pro Ser Thr Arg Leu 50 55 60 Ala Thr Gln Pro His Leu Val Gln Thr Gln Gln Val Gln Pro Gln Asn 65 70 75 80 Leu Gln Met Gln Gln Gln Asn Leu Gln Pro Ala Asn Ile Gln Gln Gln 85 90 95 Gln Ser Leu Cys Gly Leu Ser Pro Arg Arg Pro His His His His Ser 100 105 110 Arg Thr Leu Ala 115 142 99 PRT Homo sapiens 142 Trp Leu Ser Arg Leu Gln Ser Gly Met Val Pro Asn Gln Tyr Asn Pro 1 5 10 15 Leu Arg Gly Ser Val Ala Pro Gly Pro Leu Ser Thr Gln Ala Pro Ser 20 25 30 Leu Gln His Gly Met Val Gly Pro Leu His Ser Ser Leu Ala Ala Ser 35 40 45 Ala Leu Ser Gln Met Met Ser Tyr Gln Gly Leu Pro Ser Thr Arg Leu 50 55 60 Ala Thr Gln Pro His Leu Val Gln Thr Gln Gln Val Gln Pro Gln Asn 65 70 75 80 Leu Gln Met Gln Gln Gln Asn Leu Gln Pro Ala Asn Ile Gln Gln Gln 85 90 95 Gln Ser Leu 143 174 PRT Homo sapiens 143 Trp Leu Ser Arg Leu Gln Ser Gly Met Val Pro Asn Gln Tyr Asn Pro 1 5 10 15 Leu Arg Gly Ser Val Ala Pro Gly Pro Leu Ser Thr Gln Ala Pro Ser 20 25 30 Leu Gln His Gly Met Val Gly Pro Leu His Ser Ser Leu Ala Ala Ser 35 40 45 Ala Leu Ser Gln Met Met Ser Tyr Gln Gly Leu Pro Ser Thr Arg Leu 50 55 60 Ala Thr Gln Pro His Leu Val Gln Thr Gln Gln Val Gln Pro Gln Asn 65 70 75 80 Leu Gln Met Gln Gln Gln Asn Leu Gln Pro Ala Asn Ile Gln Gln Gln 85 90 95 Gln Ser Leu Gln Gln Pro His Leu Gly Val Ser Ser Ala Ala Ser Gly 100 105 110 His Leu Gly Arg Ser Phe Leu Ser Gly Glu Pro Ser Gln Ala Asp Val 115 120 125 Gln Pro Leu Gly Pro Ser Ser Leu Ala Val His Thr Ile Leu Pro Gln 130 135 140 Glu Ser Pro Ala Leu Pro Thr Ser Leu Pro Ser Ser Leu Val Pro Pro 145 150 155 160 Val Thr Ala Ala Gln Phe Leu Thr Pro Pro Ser Gln His Ser 165 170 144 297 PRT Homo sapiens 144 Trp Leu Ser Arg Leu Gln Ser Gly Met Val Pro Asn Gln Tyr Asn Pro 1 5 10 15 Leu Arg Gly Ser Val Ala Pro Gly Pro Leu Ser Thr Gln Ala Pro Ser 20 25 30 Leu Gln His Gly Met Val Gly Pro Leu His Ser Ser Leu Ala Ala Ser 35 40 45 Ala Leu Ser Gln Met Met Ser Tyr Gln Gly Leu Pro Ser Thr Arg Leu 50 55 60 Ala Thr Gln Pro His Leu Val Gln Thr Gln Gln Val Gln Pro Gln Asn 65 70 75 80 Leu Gln Met Gln Gln Gln Asn Leu Gln Pro Ala Asn Ile Gln Gln Gln 85 90 95 Gln Ser Leu Gln Pro Pro Pro Pro Pro Pro Gln Pro His Leu Gly Val 100 105 110 Ser Ser Ala Ala Ser Gly His Leu Gly Arg Ser Phe Leu Ser Gly Glu 115 120 125 Pro Ser Gln Ala Asp Val Gln Pro Leu Gly Pro Ser Ser Leu Ala Val 130 135 140 His Thr Ile Leu Pro Gln Glu Ser Pro Ala Leu Pro Thr Ser Leu Pro 145 150 155 160 Ser Ser Leu Val Pro Pro Val Thr Ala Ala Gln Phe Leu Thr Pro Pro 165 170 175 Ser Gln His Ser Tyr Ser Ser Pro Val Asp Asn Thr Pro Ser His Gln 180 185 190 Leu Gln Val Pro Glu His Pro Phe Leu Thr Pro Ser Pro Glu Ser Pro 195 200 205 Asp Gln Trp Ser Ser Ser Ser Pro His Pro Ala Phe Gln Arg Leu Arg 210 215 220 Leu Val Arg Gly Arg Leu Gln Pro Ser His Gln His Ala Val Pro Asp 225 230 235 240 Arg Pro His Ser Gly Gly Leu Gln Val Asn Gly Ala Pro His Glu Thr 245 250 255 Pro Ala Ser Phe Pro Lys Pro Ser Gly Val Cys Val Arg Ser Val Asp 260 265 270 Ala Arg Ala Asp Gln Arg Ser Leu Phe Lys Thr His Val Phe Ile Gln 275 280 285 Asn Lys Asn Glu Asp Phe Asn Phe Phe 290 295 145 49 PRT Homo sapiens 145 Ala Ala Gly Thr Leu Val Val Val Val Leu Met Pro Pro Glu Gln Leu 1 5 10 15 Arg Asn Ser Ser Phe His Phe Leu Ser Arg Glu Leu Ser Arg Val Leu 20 25 30 His Thr Asn Val Val Phe Lys Arg Asp Ala His Gly Gln Gln Met Ile 35 40 45 Phe 146 42 DNA Homo sapiens 146 cccggttggg cagcctcaac atcccctaca agatcgaggc cg 42 147 12 DNA Homo sapiens 147 atcaatgccc cc 12 148 17 DNA Homo sapiens 148 gagcacacag gccccct 17 149 14 DNA Homo sapiens 149 ttaagtagag actt 14 150 12 DNA Homo sapiens 150 gggccgtgga cg 12 151 15 DNA Homo sapiens 151 gtggaggggg gatgg 15 152 10 DNA Homo sapiens 152 gtcccctgac 10 153 10 DNA Homo sapiens 153 tcctcacccc 10 154 9 PRT Homo sapiens 154 Gly Cys His Pro Arg Trp Ser His Pro 1 5 155 13 PRT Homo sapiens 155 Pro Cys Pro Arg Arg Cys His Pro Arg Trp Ser His Pro 1 5 10 156 37 PRT Homo sapiens 156 Leu Ser Arg Asp Leu Cys Ser His Trp Ala Pro Ala Ala Trp Arg Cys 1 5 10 15 Thr Leu Phe Cys Pro Arg Arg Ala Pro Pro Cys Pro Arg Arg Cys His 20 25 30 Pro Arg Trp Ser His 35 157 34 PRT Homo sapiens 157 Cys Cys Ser His Trp Ala Pro Ala Ala Trp Arg Cys Thr Leu Phe Cys 1 5 10 15 Pro Arg Arg Ala Pro Pro Cys Pro Arg Arg Cys His Pro Arg Trp Ser 20 25 30 His Pro 158 38 PRT Homo sapiens 158 Glu Gly Arg Gly Arg Cys Ser His Trp Ala Pro Ala Ala Trp Arg Cys 1 5 10 15 Thr Leu Phe Cys Pro Arg Arg Ala Pro Pro Cys Pro Arg Arg Cys His 20 25 30 Pro Arg Trp Ser His Pro 35 159 38 PRT Homo sapiens 159 Gly Gly Gly Gly Met Gly Ser His Trp Ala Pro Ala Ala Trp Arg Cys 1 5 10 15 Thr Leu Phe Cys Pro Arg Arg Ala Pro Pro Cys Pro Arg Arg Cys His 20 25 30 Pro Arg Trp Ser His Pro 35 160 39 PRT Homo sapiens 160 Glu Arg Ala Arg Gln Thr Cys Ser His Trp Ala Pro Ala Ala Trp Arg 1 5 10 15 Cys Thr Leu Phe Cys Pro Arg Arg Ala Pro Pro Cys Pro Arg Arg Cys 20 25 30 His Pro Arg Trp Ser His Pro 35 161 40 PRT Homo sapiens 161 Arg Val Arg Ala Arg Gln Thr Cys Ser His Trp Ala Pro Ala Ala Trp 1 5 10 15 Arg Cys Thr Leu Phe Cys Pro Arg Arg Ala Pro Pro Cys Pro Arg Arg 20 25 30 Cys His Pro Arg Trp Ser His Pro 35 40 162 12 PRT Homo sapiens 162 Arg His His His His His Ser Arg Thr Leu Val Ala 1 5 10 163 70 PRT Homo sapiens 163 Asp Ser Gly Pro Ala Arg Pro Arg Ile Pro Thr Ser Pro Thr Gly Pro 1 5 10 15 Arg Ala Ser Pro Ala Leu Pro Pro Ala Cys Ser Pro Arg Ser Pro Ala 20 25 30 Phe Arg Arg Pro Ser Ser Lys Arg Arg Ala Pro Arg Asp Pro Gly Phe 35 40 45 Leu Ser Gln Ala Phe Gly Arg Leu Cys Ala Leu Cys Gly Cys Gln Gly 50 55 60 Arg Pro

Glu Glu Pro Ser 65 70 164 12 PRT Homo sapiens 164 Gly Arg Gln His Pro Gln Pro Pro Ala Thr Gly Ala 1 5 10

Claims

1. A substantially purified mutant NOTCH-1 receptor, comprising the amino acid sequence of SEQ ID NO:1 except that said amino acid sequence has one or more mutations in amino acids 1571-1622 and/or 1674-1730 and/or amino acid 2311 to amino acid 2556 such that, relative to unmutated NOTCH-1, said mutant NOTCH-1 receptor exhibits increased signaling in response to incubation with gamma-secretase.

2. The substantially purified mutant receptor of claim 1, wherein, except for said one or more mutations, said mutant receptor consists of the amino sequence of SEQ ID NO:1.

3. The substantially purified mutant NOTCH-1 receptor of claim 1, wherein said one or more mutations include one or more of the following: a) the portion of the amino acid sequence of SEQ ID NO:1 from amino acid 1571 to amino acid 1618 is replaced with a sequence selected from the group consisting of SEQ ID NO:12-SEQ ID NO:22, SEQ ID NO:102-118, and SEQ ID NO:145; b) the portion of the amino acid sequence of SEQ ID NO:1 from amino acid 1571 to amino acid 1622 is replaced with a sequence selected from the group consisting of SEQ ID NO:23- SEQ ID NO:29, or SEQ ID NO:119-127; c) the portion of the amino acid sequence of SEQ ID NO:1 from amino acid 1674 to amino acid 1730 is replaced with a sequence selected from the group consisting of SEQ ID NO:70- SEQ ID NO:75, or SEQ ID NO:128-137; d) the portion of the amino acid sequence of SEQ ID NO:1 from amino acid 2311 to amino acid 2556 is replaced with a sequence selected from the group consisting of SEQ ID NO:30- SEQ ID NO:59 or SEQ ID NO:138-144.

4. The substantially purified mutant receptor of claim 2, wherein, except for said one or more mutations, said mutant receptor consists of the amino sequence of SEQ ID NO:1.

5. The substantially purified mutant NOTCH-1 receptor of claim 1, wherein said sequence is mutated at one or more amino acid positions selected from the group consisting of: 1571; 1573; 1575; 1576; 1578; 1579; 1583; 1586; 1588; 1590; 1593; 1594; 1595; 1598; 1600-1607; 1610-1618; 1674; 1675; 1677; 1679-1684; 1686; 1695; 1696; 1703; 1706; 1715; 1720; 1725; and 1728.

6. The substantially purified receptor of claim 5, wherein, except for said one or more mutated amino acids, said receptor consists of the amino acid sequence of SEQ ID NO:1.

7. A substantially purified polynucleotide comprising a nucleotide sequence encoding the mutant NOTCH-1 receptor of claim 1.

8. The substantially purified polynucleotide of claim 7, wherein said polynucleotide consists of a nucleotide sequence encoding the mutant receptor of claim 1.

9. A substantially purified polynucleotide, at least 15 nucleotides in length which hybridizes to the polynucleotide of claim 8 under stringent conditions but which does not hybridize to a polynucleotide encoding unmutated NOTCH-1 under the same conditions.

10. A vector comprising a promoter operably linked to a polynucleotide encoding the mutant NOTCH-1 receptor of claim 1.

11. A host cell transformed with the vector of claim 10.

12. An antibody that binds to the mutant NOTCH-1 receptor of claim 1 with at least a hundredfold greater affinity than to the unmutated NOTCH-1 receptor of SEQ ID NO:1.

13. A method of diagnostically testing a biological sample for the presence of cancer cells, comprising assaying said biological sample for the mutant NOTCH-1 receptor of claim 1.

14. A method of diagnostically testing a biological sample for the presence of cancer cells, comprising assaying said biological sample for cells expressing the polynucleotide of claim 7.

15. A method of diagnostically testing a biological sample for the presence of cancer cells, comprising assaying said biological sample for cells expressing the polynucleotide of claim 8.

16. The method of claim 15, wherein said method is performed by PCR amplifying either the full length polynucleotide encoding said mutant NOTCH-1 receptor or a portion of the polynucleotide encoding said mutant NOTCH-1 receptor.

17. A method of slowing or stopping the growth of cells identified as having the mutant NOTCH-1 receptor of claim 1, comprising contacting said cells with an effective amount of a gamma-secretase inhibitor.

18. A method of treating a patient for cancer comprising: a) assaying a biological sample from said patient for the presence of cancer cells having the mutant Notch-1 receptor of claim 1; b) treating said patient with an effective amount of an agent that inhibits Notch-1 receptor signaling if said biological sample is found to have said mutant Notch-1 receptor.

19. The method of claim 18, wherein said cancer is T-cell acute lymphoblastic leukemia.

20. The method of claim 18, wherein said agent is a gamma-secretase inhibitor.

21. A method of assaying a test compound for its ability to inhibit abnormal cellular growth induced by the presence of a mutated NOTCH-1 receptor, comprising: a) incubating said test compound with cells that express the mutant NOTCH-1 receptor of claim 1, said incubation being carried out in the presence of gamma-secretase; b) comparing the amount of receptor activity that occurs as the result of the incubation of step a) with the amount of activity that occurs in a similar incubation carried out in the absence of said test compound; and c) concluding that said test compound should inhibit the growth of cells expressing mutant NOTCH-1 receptor if the amount of activity observed in the presence of said test compound is less than the amount of activity observed in the absence of said test compound.