CA2324492A1

CA2324492A1 - Secreted proteins and polynucleotides encoding them

Info

Publication number: CA2324492A1
Application number: CA002324492A
Authority: CA
Inventors: Kenneth Jacobs; John M. Mccoy; Edward R. Lavallie; Lisa A. Collins-Racie; Cheryl Evans; David Merberg; Maurice Treacy; Michael J. Agostino; Robert J. Ii Steininger
Original assignee: Individual
Current assignee: Genetics Institute LLC
Priority date: 1998-03-31
Filing date: 1999-03-30
Publication date: 1999-10-07
Also published as: EP1068307A1; WO1999050405A1; AU3457299A; JP2002509722A

Abstract

Novel polynucleotides and the proteins encoded thereby are disclosed.

Description

SECRETED PROTEINS AND POLYNUCLEOTIDES ENCODING THEM
This application is a continuation-in-part of provisional application Ser. No.
60/080,110, filed March 31,1998, which is incorporated by reference herein.
FIELD OF THE INVENTION
The present invention provides novel poly-nucleotides and proteins encoded by such polynucleotides, along with therapeutic, diagnostic and research utilities for these polynucleotides and proteins.
BACKGROUND OF THE INVENTION
Technology aimed at the discovery of protein factors (including e.g., cytokines, such as lymphokines, interferons, CSFs and interleukins) has matured rapidly over the past decade. The now routine hybridization cloning and expression cloning techniques

2 5 clone novel polynucleotides "directly" in the sense that they rely on information directly related to the discovered protein (i.e., partial DNA/amino acid sequence of the protein in the case of hybridization cloning; activity of the protein in the case of expression cloning). More recent "indirect" cloning techniques such as signal sequence cloning, which isolates DNA sequences based on the presence of a now well-recognized secretory leader

3 0 sequence motif, as well as various PCR-based or low stringency hybridization cloning techniques, have advanced the state of the art by making available large numbers of DNA/amino acid sequences for proteins that are known to have biological activity by virtue of their secreted nature in the case of leader sequence cloning, or by virtue of the cell or tissue source in the case of PCR-based techniques. It is to these proteins and the 3 5 polynucleotides encoding them that the present invention is directed.

SUMMARY OF THE INVENTION
In one embodiment, the present invention provides a composition comprising an isolated polynucleotide selected from the group consisting of:
(a) a polynucleotide comprising the nucleotide sequence of SEQ ID
NO:1;
(b) a polynucleotide comprising the nucleotide sequence of SEQ ID
NO:1 from nucleotide 216 to nucleotide 2174;
(c) a polynucleotide comprising the nucleotide sequence of SEQ ID
N0:1 from nucleotide 339 to nucleotide 2174;
(d) a polynucleotide comprising the nucleotide sequence of the full-length protein coding sequence of clone cc359 4 deposited under accession number ATCC 98715;
(e) a polynucleotide encoding the full-length protein encoded by the cDNA insert of clone cc359_4 deposited under accession number ATCC 98715;
(f) a polynucleotide comprising the nucleotide sequence of a mature protein coding sequence of clone cc359_4 deposited under accession number ATCC 98715;
(g) a polynucleotide encoding a mature protein encoded by the cDNA
insert of clone cc359 4 deposited under accession number ATCC 98715;
2 0 (h) a polynucleotide encoding a protein comprising the amino acid sequence of SEQ ID N0:2;
(i) a polynucleotide encoding a protein comprising a fragment of the amino acid sequence of SEQ ID N0:2 having biological activity, the fragment comprising eight contiguous amino acids of SEQ ID N0:2;
(j) a polynucleotide which is an allelic variant of a polynucleotide of (a)-(g) above;
(k) a polynucleotide which encodes a species homologue of the protein of (h) or (i) above ;
(1) a polynucleotide that hybridizes under stringent conditions to any 3 0 one of the polynucleotides specified in (a)-(i); and (m) a polynucleotide that hybridizes under stringent conditions to any one of the polynucleotides specified in (a)-(i) and that has a length that is at least 25% of the length of SEQ ID NO:1.

Preferably, such polynucleotide comprises the nucleotide sequence of SEQ ID
NO:1 from nucleotide 216 to nucleotide 2174; the nucleotide sequence of SEQ ID
NO:1 from nucleotide 339 to nucleotide 2174; the nucleotide sequence of the full-length protein coding sequence of clone cc359 4 deposited under accession number ATCC 98715;
or the nucleotide sequence of a mature protein coding sequence of clone cc359_4 deposited under accession number ATCC 98715. In other preferred embodiments, the polynudeotide encodes the full-length or a mature protein encoded by the cDNA
insert of clone cc359 4 deposited under accession number ATCC 98715. In further preferred embodiments, the present invention provides a polynucleotide encoding a protein comprising a fragment of the amino acid sequence of SEQ ID N0:2 having biological activity, the fragment preferably comprising eight (more preferably twenty, most preferably thirty) contiguous amino acids of SEQ ID N0:2, or a polynucleotide encoding a protein comprising a fragment of the amino acid sequence of SEQ ID N0:2 having biological activity, the fragment comprising the amino acid sequence from amino acid 321 to amino acid 330 of SEQ ID N0:2.
Other embodiments provide the gene corresponding to the cDNA sequence of SEQ
ID NO:1.
Further embodiments of the invention provide isolated polynucleoiides produced according to a process selected from the group consisting of:
2 0 (a) a process comprising the steps of:
(i) preparing one or more polynucleotide probes that hybridize in 6X SSC at 65 degrees C to a nucleotide sequence selected from the group consisting of:
(aa) SEQ ID NO:1, but excluding the poly(A) tail at the 2 5 3' end of SEQ ID NO:1; and (ab) the nucleotide sequence of the cDNA insert of clone cc359_4 deposited under accession number ATCC 98715;
(ii) hybridizing said probes) to human genomic DNA in conditions at least as stringent as 4X SSC at 50 degrees C; and 3 0 (iii) isolating the DNA polynucleotides detected with the probe(s);
and (b) a process comprising the steps of:

(i) preparing one or more polynucleotide primers that hybridize in 6X SSC at 65 degrees C to a nucleotide sequence selected from the group consisting of:
(ba) SEQ ID NO:1, but excluding the poly(A) tail at the 3' end of SEQ ID N0:1; and (bb) the nucleotide sequence of the cDNA insert of clone cc359 4 deposited under accession number ATCC 98715;
(ii) hybridizing said primers) to human genomic DNA in conditions at least as stringent as 4X SSC at 50 degrees C;
I 0 (iii) amplifying human DNA sequences; and (iv) isolating the polynucleotide products of step (b)(iii).
Preferably the polynucleotide isolated according to the above process comprises a nucleotide sequence corresponding to the cDNA sequence of SEQ ID NO:1, and extending contiguously from a nucleotide sequence corresponding to the 5' end of SEQ ID
NO:1 to a nucleotide sequence corresponding to the 3' end of SEQ ID NO:1 , but excluding the poly(A) tail at the 3' end of SEQ ID NO:1. Also preferably the polynucleotide isolated according to the above process comprises a nucleotide sequence corresponding to the cDNA sequence of SEQ FD NO:1 from nucleotide 216 to nucleotide 2174, and extending contiguously from a nucleotide sequence corresponding to the 5' end of said sequence of 2 0 SEQ ID NO:1 from nucleotide 216 to nucleotide 2174, to a nucleotide sequence corresponding to the 3' end of said sequence of SEQ ID NO:1 from nucleotide 216 to nucleotide 2174. Also preferably the polynucleotide isolated according to the above process comprises a nucleotide sequence corresponding to the cDNA sequence of SEQ ID
NO:1 from nucleotide 339 to nucleotide 2174, and extending contiguously from a 2 5 nucleotide sequence corresponding to the 5' end of said sequence of SEQ ID
NO:1 from nucleotide 339 to nucleotide 2174, to a nucleotide sequence corresponding to the 3' end of said sequence of SEQ ID NO:1 from nucleotide 339 to nucleotide 2174.
In other embodiments, the present invention provides a composition comprising a protein, wherein said protein comprises an amino acid sequence selected from the group 3 0 consisting of:
(a) the amino acid sequence of SEQ ID N0:2;
(b) a fragment of the amino acid sequence of SEQ ID N0:2, the fragment comprising eight contiguous amino acids of SEQ ID N0:2; and

4

5 PCTNS99/06946 (c) the amino acid sequence encoded by the cDNA insert of clone cc359 4 deposited under accession number ATCC 98715;
the protein being substantially free from other mammalian proteins. Preferably such protein comprises the amino acid sequence of SEQ ID N0:2. In further preferred embodiments, the present invention provides a protein comprising a fragment of the amino acid sequence of SEQ ID N0:2 having biological activity, the fragment preferably comprising eight (more preferably twenty, most preferably thirty) contiguous amino acids of SEQ ID N0:2, or a protein comprising a fragment of the amino acid sequence of SEQ
ID N0:2 having biological activity, the fragment comprising the amino acid sequence from amino acid 321 to amino acid 330 of SEQ ID N0:2.
In one embodiment, the present invention provides a composition comprising an isolated polynucleotide selected from the group consisting of:
(a) a polynudeotide comprising the nucleotide sequence of SEQ ID
N0:3;
(b) a polynucleotide comprising the nucleotide sequence of SEQ ID
N0:3 from nucleotide 202 to nucleotide 414;
(c) a polynucleotide comprising the nucleotide sequence of SEQ ID
N0:3 from nucleotide 370 to nucleotide 414;
(d) a polynucleotide comprising the nucleotide sequence of the full-2 0 length protein coding sequence of clone ct547_2 deposited under accession number ATCC 98715;
(e) a polynucleotide encoding the full-length protein encoded by the cDNA insert of clone ct547 2 deposited under accession number ATCC 98715;
(f) a polynucleotide comprising the nucleotide sequence of a mature 2 5 protein coding sequence of clone ct547_2 deposited under accession number ATCC 98715;
(g) a polynucleotide encoding a mature protein encoded by the cDNA
insert of clone ct547 2 deposited under accession number ATCC 98715;
(h) a polynucleotide encoding a protein comprising the amino acid 3 0 sequence of SEQ ID N0:4;
(i) a polynucleotide encoding a protein comprising a fragment of the amino acid sequence of SEQ ID N0:4 having biological activity, the fragment comprising eight contiguous amino acids of SEQ ID N0:4;

(j) a polynucleotide which is an allelic variant of a polynucleotide of (a)-(g) above;
(k) a polynucleotide which encodes a species homologue of the protein of (h) or (i) above ;
(1) a polynucleotide that hybridizes under stringent conditions to any one of the polynucleotides specified in (a)-(i); and (m) a polynudeotide that hybridizes under stringent conditions to any one of the polynucleotides specified in (a)-(i) and that has a length that is at least 25% of the length of SEQ ID N0:3.
Preferably, such polynucleotide comprises the nucleotide sequence of SEQ ID
N0:3 from nucleotide 202 to nucleotide 414; the nucleotide sequence of SEQ ID
N0:3 from nucleotide 370 to nucleotide 414; the nucleotide sequence of the full-length protein coding sequence of clone ct547 2 deposited under accession number ATCC 98715; or the nucleotide sequence of a mature protein coding sequence of clone ct547_2 deposited under accession number ATCC 98715. In other preferred embodiments, the polynucleotide encodes the full-length or a mature protein encoded by the cDNA
insert of clone ct547 2 deposited under accession number ATCC 98715. In further preferred embodiments, the present invention provides a polynucleotide encoding a protein comprising a fragment of the amino acid sequence of SEQ ID N0:4 having biological 2 0 activity, the fragment preferably comprising eight (more preferably twenty, most preferably thirty) contiguous amino acids of SEQ ID N0:4, or a polynucleotide encoding a protein comprising a fragment of the amino acid sequence of SEQ ID N0:4 having biological activity, the fragment comprising the amino acid sequence from amino acid 30 to amino acid 39 of SEQ ID N0:4.
2 5 Other embodiments provide the gene corresponding to the cDNA sequence of SEQ
ID N0:3.
Further embodiments of the invention provide isolated polynucleotides produced according to a process selected from the group consisting of:
(a) a process comprising the steps of:
3 0 (i) preparing one or more polynucleotide probes that hybridize in 6X SSC at 65 degrees C to a nucleotide sequence selected from the group consisting of:
(aa) SEQ ID N0:3, but excluding the poly(A) tail at the 3' end of SEQ ID N0:3; and

6 (ab) the nucleotide sequence of the cDNA insert of clone ct5~47_2 deposited under accession number ATCC 98715;
(ii) hybridizing said probes) to human genomic DNA in conditions at least as stringent as 4X SSC at 50 degrees C; and (iii) isolating the DNA polynucleotides detected with the probe(s);
and (b) a process comprising the steps of:
(i) preparing one or more polynucleotide primers that hybridize in 6X SSC at 65 degrees C to a nucleotide sequence selected from the group consisting of:
(ba) SEQ ID N0:3, but excluding the poly(A) tail at the 3' end of SEQ ID N0:3; and (bb) the nucleotide sequence of the cDNA insert of clone ct547_2 deposited under accession number ATCC 98715;
(ii) hybridizing said primers) to human genomic DNA in conditions at least as stringent as 4X SSC at 50 degrees C;
(iii) amplifying human DNA sequences; and (iv) isolating the polynucleotide products of step (b)(iii).
2 0 Preferably the polynucleotide isolated according to the above process comprises a nucleotide sequence corresponding to the cDNA sequence of SEQ ID N0:3, and extending contiguously from a nucleotide sequence corresponding to the 5' end of SEQ ID
N0:3 to a nucleotide sequence corresponding to the 3' end of SEQ ID N0:3 , but excluding the poly(A) tail at the 3' end of SEQ ID N0:3. Also preferably the polynucleotide isolated 2 5 according to the above process comprises a nucleotide sequence corresponding to the cDNA sequence of SEQ ID N0:3 from nucleotide 202 to nucleotide 414, and extending contiguously from a nucleotide sequence corresponding to the 5' end of said sequence of SEQ ID N0:3 from nucleotide 202 to nucleotide 414, to a nucleotide sequence corresponding to the 3' end of said sequence of SEQ ID N0:3 from nucleotide 202 to 3 0 nucleotide 414. Also preferably the polynucleotide isolated according to the above process comprises a nucleotide sequence corresponding to the cDNA sequence of SEQ ID
N0:3 from nucleotide 370 to nucleotide 414, and extending contiguously from a nucleotide sequence corresponding to the 5' end of said sequence of SEQ ID
N0:3 from

7 nucleotide 370 to nucleotide 414, to a nucleotide sequence corresponding to the 3' end of said sequence of SEQ ID N0:3 from nucleotide 370 to nucleotide 414.
In other embodiments, the present invention provides a composition comprising a protein, wherein said protein comprises an amino acid sequence selected from the group consisting of:
(a) the amino acid sequence of SEQ ID N0:4;
(b) a fragment of the amino acid sequence of SEQ ID N0:4, the fragment comprising eight contiguous amino acids of SEQ ID N0:4; and (c) the amino acid sequence encoded by the cDNA insert of clone ct547 2 deposited under accession number ATCC 98715;
the protein being substantially free from other mammalian proteins. Preferably such protein comprises the amino acid sequence of SEQ ID N0:4. In further preferred embodiments, the present invention provides a protein comprising a fragment of the amino acid sequence of SEQ ID N0:4 having biological activity, the fragment preferably comprising eight (more preferably twenty, most preferably thirty) contiguous amino acids of SEQ ID N0:4, or a protein comprising a fragment of the amino acid sequence of SEQ
ID N0:4 having biological activity, the fragment comprising the amino acid sequence from amino acid 30 to amino acid 39 of SEQ ID NO:4.
In one embodiment, the present invention provides a composition comprising an 2 0 isolated polynucleotide selected from the group consisting of:
(a) a polynucleotide comprising the nucleotide sequence of SEQ ID
N0:5;
(b) a polynucleotide comprising the nucleotide sequence of SEQ ID
N0:5 from nucleotide 206 to nucleotide 415;
2 5 (c) a polynucleotide comprising the nucleotide sequence of SEQ ID
N0:5 from nucleotide 293 to nucleotide 415;
(d) a polynucleotide comprising the nucleotide sequence of the full-length protein coding sequence of clone en553_1 deposited under accession number ATCC 98715;
3 0 (e) a polynucleotide encoding the full-length protein encoded by the cDNA insert of clone en553_1 deposited under accession number ATCC 98715;
(f) a polynucleotide comprising the nucleotide sequence of a mature protein coding sequence of clone en553_1 deposited under accession number ATCC 98715;

8 WO 99/50405 PC'TIUS99/06946 (g) a polynucleotide encoding a mature protein encoded by the cDNA
insert of clone en553_I deposited under accession number ATCC 98715;
(h) a polynucleotide encoding a protein comprising the amino acid sequence of SEQ ID N0:6;
(i) a polynucleotide encoding a protein comprising a fragment of the amino acid sequence of SEQ ID N0:6 having biological activity, the fragment comprising eight contiguous amino acids of SEQ ID N0:6;
(j) a polynucleotide which is an allelic variant of a polynucleotide of (a)-(g) above;
(k) a polynucleotide which encodes a species homologue of the protein of (h) or (i) above ;
(1) a polynucleotide that hybridizes under stringent conditions to any one of the polynucleotides specified in (a)-(i); and (m) a polynucleotide that hybridizes under stringent conditions to any one of the polynucleotides specified in (a)-(i) and that has a length that is at least 25% of the length of SEQ ID N0:5.
Preferably, such polynucleodde comprises the nucleotide sequence of SEQ ID
N0:5 from nucleotide 206 to nucleotide 415; the nucleotide sequence of SEQ ID
N0:5 from nucleotide 293 to nucleotide 415; the nucleotide sequence of the full-length protein coding 2 0 sequence of clone en553_1 deposited under accession number ATCC 98715; or the nucleotide sequence of a mature protein coding sequence of clone en553_1 deposited under accession number ATCC 98715. In other preferred embodiments, the polynucleotide encodes the full-length or a mature protein encoded by the cDNA
insert of clone en553_1 deposited under accession number ATCC 98715. In further preferred 2 5 embodiments, the present invention provides a polynucleotide encoding a protein comprising a fragment of the amino acid sequence of SEQ ID NO:6 having biological activity, the fragment preferably comprising eight (more preferably twenty, most preferably thirty) contiguous amino acids of SEQ ID N0:6, or a polynucleotide encoding a protein comprising a fragment of the amino acid sequence of SEQ ID N0:6 having 3 0 biological activity, the fragment comprising the amino acid sequence from amino acid 30 to amino acid 39 of SEQ ID N0:6.
Other embodiments provide the gene corresponding to the cDNA sequence of SEQ
ID N0:5.

9 Further embodiments of the invention provide isolated polynudeotides produced according to a process selected from the group consisting of:
(a) a process comprising the steps of:
(i) preparing one or more polynucleotide probes that hybridize in 6X SSC at 65 degrees C to a nucleotide sequence selected from the group consisting of:
(aa) SEQ ID N0:5, but excluding the poly(A) tail at the 3' end of SEQ ID N0:5; and (ab) the nucleotide sequence of the cDNA insert of clone en553_1 deposited under accession number ATCC 98715;
(ii) hybridizing said probes) to human genomic DNA in conditions at least as stringent as 4X SSC at 50 degrees C; and (iii) isolating the DNA polynucleotides detected with the probe(s);
and (b) a process comprising the steps of:
(i) preparing one or more polynucleotide primers that hybridize in 6X SSC at 65 degrees C to a nucleotide sequence selected from the group consisting of:
2 0 (ba) SEQ ID NO:S, but excluding the poly(A) tail at the 3' end of SEQ ID N0:5; and (bb) the nucleotide sequence of the cDNA insert of clone en553_1 deposited under accession number ATCC 98715;
(ii) hybridizing said primers) to human genomic DNA in 2 5 conditions at least as stringent as 9X SSC at 50 degrees C;
(iii) amplifying human DNA sequences; and (iv) isolating the polynucleotide products of step (b)(iii).
Preferably the polynucleotide isolated according to the above process comprises a nucleotide sequence corresponding to the cDNA sequence of SEQ ID N0:5, and extending 3 0 contiguously from a nucleotide sequence corresponding to the 5' end of SEQ
ID N0:5 to a nucleotide sequence corresponding to the 3' end of SEQ ID N0:5 , but excluding the poly(A} tail at the 3' end of SEQ ID N0:5. Also preferably the polynucleotide isolated according to the above process comprises a nucleotide sequence corresponding to the cDNA sequence of SEQ ID N0:5 from nucleotide 206 to nucleotide 415, and extending contiguously from a nucleotide sequence corresponding to the 5' end of said sequence of SEQ ID N0:5 from nucleotide 206 to nucleotide 415, to a nucleotide sequence corresponding to the 3' end of said sequence of SEQ ID N0:5 from nucleotide 206 to nucleotide 415. Also preferably the polynucleotide isolated according to the above process comprises a nucleotide sequence corresponding to the cDNA sequence of SEQ ID
N0:5 from nucleotide 293 to nucleotide 415, and extending contiguously from a nucleotide sequence corresponding to the 5' end of said sequence of SEQ ID
N0:5 from nucleotide 293 to nucleotide 415, to a nucleotide sequence corresponding to the 3' end of said sequence of SEQ ID N0:5 from nucleotide 293 to nucleotide 415.
In other embodiments, the present invention provides a composition comprising a protein, wherein said protein comprises an amino acid sequence selected from the group consisting of:
(a) the amino acid sequence of SEQ ID N0:6;
(b) a fragment of the amino acid sequence of SEQ ID N0:6, the fragment comprising eight contiguous amino acids of SEQ ID N0:6; and (c) the amino acid sequence encoded by the cDNA insert of clone en553_1 deposited under accession number ATCC 98715;
the protein being substantially free from other mammalian proteins. Preferably such protein comprises the amino acid sequence of SEQ ID N0:6. In further preferred 2 0 embodiments, the present invention provides a protein comprising a fragment of the amino acid sequence of SEQ ID N0:6 having biological activity, the fragment preferably comprising eight (more preferably twenty, most preferably thirty) contiguous amino ands of SEQ ID N0:6, or a protein comprising a fragment of the amino acid sequence of SEQ
ID N0:6 having biological activity, the fragment comprising the amino acid sequence from 2 5 amino acid 30 to amino acid 39 of SEQ ID N0:6.
In one embodiment, the present invention provides a composition comprising an isolated polynucleotide selected from the group consisting of:
(a) a polynucleotide comprising the nucleotide sequence of SEQ ID
N0:7;
3 0 (b) a polynucleotide comprising the nucleotide sequence of SEQ ID
N0:7 from nucleotide 120 to nucleotide 1202;
(c) a polynucleotide comprising the nucleotide sequence of the full-length protein coding sequence of clone nn296_2 deposited under accession number ATCC 98715;
lI

WO 99!50405 PCT/US99/06946 (d) a polynucleotide encoding the full-length protein encoded by the cDNA insert of clone nn296 2 deposited under accession number ATCC 98715;
(e) a polynucleotide comprising the nucleotide sequence of a mature protein coding sequence of clone nn296_2 deposited under accession number ATCC 98715;
(f) a polynucleotide encoding a mature protein encoded by the cDNA
insert of clone nn296 2 deposited under accession number ATCC 98715;
(g) a polynucleotide encoding a protein comprising the amino acid sequence of SEQ ID N0:8;
2 0 (h) a polynucleotide encoding a protein comprising a fragment of the amino acid sequence of SEQ ID N0:8 having biological activity, the fragment comprising eight contiguous amino acids of SEQ ID N0:8;
(i) a polynucleotide which is an allelic variant of a polynucleotide of (a)-(f) above;
(j) a polynucleotide which encodes a species homologue of the protein of (g) or (h) above ;
(k) a polynucleotide that hybridizes under stringent conditions to any one of the polynucleotides specified in (a)-(h); and (1) a polynucleotide that hybridizes under stringent conditions to any 2 0 one of the polynucleotides specified in (a)-(h) and that has a length that is at least 25% of the length of SEQ ID N0:7.
Preferably, such polynucleotide comprises the nucleotide sequence of SEQ ID
N0:7 from nucleotide 120 to nucleotide 1202; the nucleotide sequence of the full-length protein coding sequence of clone nn296_2 deposited under accession number ATCC
2 5 98715; or the nucleotide sequence of a mature protein coding sequence of clone nn296_2 deposited under accession number ATCC 98715. In other preferred embodiments, the polynucleotide encodes the full-length or a mature protein encoded by the cDNA
insert of clone nn296 2 deposited under accession number ATCC 98715. In further preferred embodiments, the present invention provides a polynucleotide encoding a protein 3 0 comprising a fragment of the amino acid sequence of SEQ ID N0:8 having biological activity, the fragment preferably comprising eight (more preferably twenty, most preferably thirty) contiguous amino acids of SEQ ID N0:8, or a polynucleotide encoding a protein comprising a fragment of the amino acid sequence of SEQ ID N0:8 having biological activity, the fragment comprising the amino acid sequence from amino and I75 to amino acid 184 of SEQ ID N0:8.
Other embodiments provide the gene corresponding to the cDNA sequence of SEQ
ID N0:7.
Further embodiments of the invention provide isolated polynucleotides produced according to a process selected from the group consisting of:
(a) a process comprising the steps of:
(i) preparing one or more polynucleotide probes that hybridize in 6X SSC at 65 degrees C to a nucleotide sequence selected from the group consisting of:
(aa) SEQ ID N0:7, but excluding the poly(A) tail at the 3' end of SEQ ID N0:7; and (ab) the nucleotide sequence of the cDNA insert of clone nn296_2 deposited under accession number ATCC 98715;
(ii) hybridizing said probes) to human genomic DNA in conditions at least as stringent as 4X SSC at 50 degrees C; and (iii) isolating the DNA polynucleotides detected with the probe(s);
and 2 0 (b) a process comprising the steps of:
(i) preparing one or more polynucleotide primers that hybridize in 6X SSC at 65 degrees C to a nucleotide sequence selected from the group consisting of:
(ba) SEQ ID N0:7, but excluding the poly(A) tail at the 2 5 3' end of SEQ ID N0:7; and (bb) the nucleotide sequence of the cDNA insert of clone nn296_2 deposited under accession number ATCC 98715;
(ii) hybridizing said primers) to human genomic DNA in conditions at least as stringent as 4X SSC at 50 degrees C;
3 0 (iii) amplifying human DNA sequences; and (iv) isolating the polynucleotide products of step (b)(iii).
Preferably the polynucleotide isolated according to the above process comprises a nucleotide sequence corresponding to the cDNA sequence of SEQ ID N0:7, and extending contiguously from a nucleotide sequence corresponding to the 5' end of SEQ ID
N0:7 to a nucleotide sequence corresponding to the 3' end of SEQ ID N0:7 , but excluding the poly(A) tail at the 3' end of SEQ ID N0:7. Also preferably the polynucleotide isolated according to the above process comprises a nucleotide sequence corresponding to the cDNA sequence of SEQ ID N0:7 from nucleotide 120 to nucleotide 1202, and extending contiguously from a nucleotide sequence corresponding to the 5' end of said sequence of SEQ ID N0:7 from nucleotide 120 to nucleotide 1202, to a nucleotide sequence corresponding to the 3' end of said sequence of SEQ ID N0:7 from nucleotide 120 to nucleotide 1202.
In other embodiments, the present invention provides a composition comprising a protein, wherein said protein comprises an amino acid sequence selected from the group consisting of:
(a) the amino acid sequence of SEQ ID N0:8;
(b) a fragment of the amino acid sequence of SEQ ID NO:B, the fragment comprising eight contiguous amino acids of SEQ ID N0:8; and (c) the amino acid sequence encoded by the cDNA insert of clone nn296 2 deposited under accession number ATCC 98715;
the protein being substantially free from other mammalian proteins. Preferably such protein comprises the amino acid sequence of SEQ ID N0:8. In further preferred embodiments, the present invention provides a protein comprising a fragment of the 2 0 amino acid sequence of SEQ ID N0:8 having biological activity, the fragment preferably comprising eight (more preferably twenty, most preferably thirty) contiguous amino ands of SEQ ID N0:8, or a protein comprising a fragment of the amino acid sequence of SEQ
ID N0:8 having biological activity, the fragment comprising the amino acid sequence from amino acid 175 to amino acid 184 of SEQ ID N0:8.
2 5 In one embodiment, the present invention provides a composition comprising an isolated polynucleotide selected from the group consisting of:
(a) a polynucleotide comprising the nucleotide sequence of SEQ ID
N0:9;
(b) a polynucleotide comprising the nucleotide sequence of SEQ ID
3 0 N0:9 from nucleotide 597 to nucleotide 704;
(c} a polynucleotide comprising the nucleotide sequence of the full-length protein coding sequence of clone nq27_13 deposited under accession number ATCC 98715;

(d) a polynucleotide encoding the full-length protein encoded by the cDNA insert of clone nq27_13 deposited under accession number ATCC 98715;
(e) a polynucleotide comprising the nucleotide sequence of a mature protein coding sequence of clone nq27_13 deposited under accession number ATCC 98715;
(f) a polynucleotide encoding a mature protein encoded by the cDNA
insert of clone nq27_13 deposited under accession number ATCC 98715;
(g) a polynucleotide encoding a protein comprising the amino acid sequence of SEQ ID N0:10;
(h) a polynucleotide encoding a protein comprising a fragment of the amino acid sequence of SEQ ID NO:10 having biological activity, the fragment comprising eight contiguous amino acids of SEQ ID NO:10;
(i) a polynucleotide encoding a protein comprising the amino acid sequence of SEQ ID N0:32;
(j) a polynucleotide encoding a protein comprising a fragment of the amino acid sequence of SEQ ID N0:32 having biological activity, the fragment comprising eight contiguous amino acids of SEQ ID N0:32;
(k) a polynucleotide which is an allelic variant of a polynucleotide of (a)-(f) above;
2 0 (1) a polynucleotide which encodes a species homologue of the protein of (g)-(j) above ;
(m) a polynucleotide that hybridizes under stringent conditions to any one of the polynucleotides specified in (a)-(j); and (n) a polynucleotide that hybridizes under stringent conditions to any 2 5 one of the polyriucleotides specified in (a)-(j) and that has a length that is at least 25% of the length of SEQ ID N0:9.
Preferably, such polynucleotide comprises the nucleotide sequence of SEQ ID
N0:9 from nucleotide 597 to nucleotide 704; the nucleotide sequence of the full-length protein coding sequence of clone nq27_13 deposited under accession number ATCC
3 0 98715; or the nucleotide sequence of a mature protein coding sequence of clone nq27 13 deposited under accession number ATCC 98715. In other preferred embodiments, the polynucleotide encodes the full-length or a mature protein encoded by the cDNA
insert of clone nq27_13 deposited under accession number ATCC 98715. In further preferred embodiments, the present invention provides a polynucleotide encoding a protein comprising a fragment of the amino acid sequence of SEQ ID NO:10 having biological activity, the fragment preferably comprising eight (more preferably twenty, most ' preferably thirty) contiguous amino acids of SEQ ID NO:10, or a polynucleotide encoding a protein comprising a fragment of the amino acid sequence of SEQ ID NO:10 having biological activity, the fragment comprising the amino acid sequence from amino acid 13 to amino acid 22 of SEQ ID NO:20.
Other embodiments provide the gene corresponding to the cDNA sequence of SEQ
ID N0:9.
Further embodiments of the invention provide isolated polynucleotides produced according to a process selected from the group consisting of:
(a) a process comprising the steps of:
(i) preparing one or more polynucleotide probes that hybridize in 6X SSC at 65 degrees C to a nucleotide sequence selected from the group consisting of:
(aa) SEQ ID N0:9, but excluding the poly(A) tail at the 3' end of SEQ ID N0:9; and (ab) the nucleotide sequence of the cDNA insert of clone nq27_13 deposited under accession number ATCC 98715;
(ii) hybridizing said probes) to human genomic DNA in 2 0 conditions at least as stringent as 4X SSC at 50 degrees C; and (iii) isolating the DNA polynucleotides detected with the probe(s);
and (b) a process comprising the steps of:
2 5 (i) preparing one or more polynucleotide primers that hybridize in 6X SSC at 65 degrees C to a nucleotide sequence selected from the group consisting of:
(ba) SEQ ID N0:9, but excluding the poly(A) tail at the 3' end of SEQ ID N0:9; and 3 0 (bb) the nucleotide sequence of the cDNA insert of clone nq27_13 deposited under accession number ATCC 98715;
(ii) hybridizing said primers) to human genomic DNA in conditions at least as stringent as 9X SSC at 50 degrees C;
(iii) amplifying human DNA sequences; and (iv) isolating the polynucleotide products of step (b)(iii).
Preferably the polynucleotide isolated according to the above process comprises a nucleotide sequence corresponding to the cDNA sequence of SEQ ID N0:9, and extending contiguously from a nucleotide sequence corresponding to the 5' end of SEQ ID
N0:9 to a nucleotide sequence corresponding to the 3' end of SEQ ID N0:9 , but excluding the poly(A) tail at the 3' end of SEQ ID NO:9. Also preferably the polynucleotide isolated according to the above process comprises a nucleotide sequence corresponding to the cDNA sequence of SEQ ID N0:9 from nucleotide 597 to nucleotide 704, and extending contiguously from a nucleotide sequence corresponding to the 5' end of said sequence of SEQ ID N0:9 from nucleotide 597 to nucleotide 704, to a nucleotide sequence corresponding to the 3' end of said sequence of SEQ ID N0:9 from nucleotide 597 to nucleotide 704.
In other embodiments, the present invention provides a composition comprising a protein, wherein said protein comprises an amino acid sequence selected from the group consisting of:
(a) the amino acid sequence of SEQ ID NO:10;
(b) a fragment of the amino acid sequence of SEQ ID NO:10, the fragment comprising eight contiguous amino acids of SEQ ID NO:10; and (c) the amino acid sequence of SEQ ID N0:32;
2 0 (d) a fragment of the amino acid sequence of SEQ ID N0:32, the fragment comprising eight contiguous amino acids of SEQ ID N0:32; and (e) the amino acid sequence encoded by the cDNA insert of clone nq27_13 deposited under accession number ATCC 98715;
the protein being substantially free from other mammalian proteins. Preferably such 2 5 protein comprises the amino acid sequence of SEQ ID NO:10. In further preferred embodiments, the present invention provides a protein comprising a fragment of the amino acid sequence of SEQ ID NO:10 having biological activity, the fragment preferably comprising eight (more preferably twenty, most preferably thirty) contiguous amino acids of SEQ ID NO:10, or a protein comprising a fragment of the amino acid sequence of SEQ
3 0 ID N0:10 having biological activity, the fragment comprising the amino acid sequence from amino acid 13 to amino acid 22 of SEQ ID N0:10.
In one embodiment, the present invention provides a composition comprising an isolated polynucleotide selected from the group consisting of:

(a) a polynucleotide comprising the nucleotide sequence of SEQ ID
N0:11;
(b) a polynucleotide comprising the nucleotide sequence of SEQ ID
NO:11 from nucleotide 44 to nucleotide 475;
(c) a polynucleotide comprising the nucleotide sequence of the full-length protein coding sequence of clone pk65 4 deposited under accession number ATCC 98715;
(d) a polynucleotide encoding the full-length protein encoded by the cDNA insert of clone pk65_4 deposited under accession number ATCC 98715;
(e) a polynucleotide comprising the nucleotide sequence of a mature protein coding sequence of clone pk65 4 deposited under accession number ATCC 98715;
(f) a polynucleotide encoding a mature protein encoded by the cDNA
insert of clone pk65 4 deposited under accession number ATCC 98715;
(g) a polynucleotide encoding a protein comprising the amino acid sequence of SEQ ID N0:12;
(h) a polynucleotide encoding a protein comprising a fragment of the amino acid sequence of SEQ ID N0:12 having biological activity, the fragment comprising eight contiguous amino acids of SEQ ID N0:12;
2 0 (i) a polynucleotide which is an allelic variant of a polynucleotide of (a)-(f) above;
(j) a polynucleotide which encodes a species homologue of the protein of (g) or (h) above ;
(k) a polynucleotide that hybridizes under stringent conditions to any 2 5 one of the polynucleotides specified in (a~(h); and (1) a polynucleotide that hybridizes under stringent conditions to any one of the polynucleotides specified in (a)-(h) and that has a length that is at least 25% of the length of SEQ ID N0:11.
Preferably, such polynucleotide comprises the nucleotide sequence of SEQ ID
3 0 NO:11 from nucleotide 44 to nucleotide 475; the nucleotide sequence of the full-length protein coding sequence of clone pk65_4 deposited under accession number ATCC
98715;
or the nucleotide sequence of a mature protein coding sequence of clone pk65 4 deposited under accession number ATCC 98715. In other preferred embodiments, the polynucleotide encodes the full-length or a mature protein encoded by the cDNA
insert of clone pk65 4 deposited under accession number ATCC 98715. In further preferred embodiments, the present invention provides a polynucleotide encoding a protein comprising a fragment of the amino acid sequence of SEQ ID N0:12 having biological activity, the fragment preferably comprising eight (more preferably twenty, most preferably thirty) contiguous amino acids of SEQ ID NO:12, or a polynucleotide encoding a protein comprising a fragment of the amino acid sequence of SEQ ID N0:12 having biological activity, the fragment comprising the amino acid sequence from amino acid 67 to amino acid 76 of SEQ ID N0:12.
Other embodiments provide the gene corresponding to the cDNA sequence of SEQ
ID NO:11.
Further embodiments of the invention provide isolated polynucleotides produced according to a process selected from the group consisting of:
(a) a process comprising the steps of:
(i) preparing one or more polynucleotide probes that hybridize in 6X SSC at 65 degrees C to a nucleotide sequence selected from the group consisting of:
(aa) SEQ ID N0:11, but excluding the poly(A) tail at the 3' end of SEQ ID NO:11; and (ab) the nucleotide sequence of the cDNA insert of clone 2 0 pk65_4 deposited under accession number ATCC 98715;
(ii) hybridizing said probes) to human genomic DNA in conditions at least as stringent as 4X SSC at 50 degrees C; and (iii) isolating the DNA polynucleotides detected with the probe(s);
2 5 and (b) a process comprising the steps of:
(i) preparing one or more polynucleotide primers that hybridize in 6X SSC at 65 degrees C to a nucleotide sequence selected from the group consisting of:
3 0 (ba) SEQ ID NO:11, but excluding the poly(A) tail at the 3' end of SEQ ID N0:11; and (bb) the nucleotide sequence of the cDNA insert of clone pk65_4 deposited under accession number ATCC 98715;

WO 99/50405 ~ PCT/US99/06946 (ii) hybridizing said prirner(s) to human genomic DNA in conditions at least as stringent as 4X SSC at 50 degrees C;
(iii) amplifying human DNA sequences; and (iv) isolating the polynucleotide products of step (b)(iii).
Preferably the polynucleotide isolated according to the above process comprises a nucleotide sequence corresponding to the cDNA sequence of SEQ ID NO:11, and extending contiguously from a nucleotide sequence corresponding to the 5' end of SEQ
ID N0:11 to a nucleotide sequence corresponding to the 3' end of SEQ ID NO:11 , but excluding the poly(A) tail at the 3' end of SEQ ID NO:11. Also preferably the polynucleotide isolated according to the above process comprises a nucleotide sequence corresponding to the cDNA sequence of SEQ ID N0:11 from nucleotide 44 to nucleotide 475, and extending contiguously from a nucleotide sequence corresponding to the 5' end of said sequence of SEQ ID N0:11 from nucleotide 44 to nucleotide 475, to a nucleotide sequence corresponding to the 3' end of said sequence of SEQ ID NO:11 from nucleotide 44 to nucleotide 475.
In other embodiments, the present invention provides a composition comprising a protein, wherein said protein comprises an amino acid sequence selected from the group consisting of:
(a) the amino acid sequence of SEQ ID N0:12;
2 0 (b) a fragment of the amino acid sequence of SEQ ID N0:12, the fragment comprising eight contiguous amino acids of SEQ ID N0:12; and (c) the amino acid sequence encoded by the cDNA insert of clone pk65 4 deposited under accession number ATCC 98715;
the protein being substantially free from other mammalian proteins. Preferably such 2 5 protein comprises the amino acid sequence of SEQ ID N0:12. In further preferred embodiments, the present invention provides a protein comprising a fragment of the amino acid sequence of SEQ ID N0:12 having biological activity, the fragment preferably comprising eight (more preferably twenty, most preferably thirty) contiguous amino acids of SEQ ID N0:12, or a protein comprising a fragment of the amino acid sequence of SEQ
3 0 ID N0:12 having biological activity, the fragment comprising the amino acid sequence from amino acid 67 to amino acid 76 of SEQ ID N0:12.
In one embodiment, the present invention provides a composition comprising an isolated polynucleotide selected from the group consisting of:

(a) a polynucleotide comprising the nucleotide sequence of SEQ ID
N0:13;
(b} a polynucleotide comprising the nucleotide sequence of SEQ ID
N0:13 from nucleotide 285 to nucleotide 590;
(c) a polynucleotide comprising the nucleotide sequence of SEQ ID
N0:13 from nucleotide 408 to nucleotide 590;
(d) a polynucleotide comprising the nucleotide sequence of the full-length protein coding sequence of clone pk855_1 deposited under accession number ATCC 98715;
(e) a polynucleotide encoding the full-length protein encoded by the cDNA insert of clone pk855_1 deposited under accession number ATCC 98715;
(f) a polynucleotide comprising the nucleotide sequence of a mature protein coding sequence of clone pk855_1 deposited under accession number ATCC 98715;
(g) a polynudeotide encoding a mature protein encoded by the cDNA
insert of clone pk855_1 deposited under accession number ATCC 98715;
(h) a polynucleotide encoding a protein comprising the amino acid sequence of SEQ ID N0:14;
(i) a polynucleotide encoding a protein comprising a fragment of the 2 0 amino acid sequence of SEQ ID N0:14 having biological activity, the fragment comprising eight contiguous amino acids of SEQ ID N0:14;
(j) a polynucleotide which is an allelic variant of a polynucleotide of (a}-(g) above;
(k) a polynucleotide which encodes a species homologue of the protein 2 5 of (h) or (i) above ;
(1) a polynucleotide that hybridizes under stringent conditions to any one of the polynucleotides specified in (a)-(i); and (m) a polynucleotide that hybridizes under stringent conditions to any one of the polynucleotides specified in (a)-(i) and that has a length that is at least 3 0 25% of the length of SEQ ID N0:13.
Preferably, such polynucleotide comprises the nucleotide sequence of SEQ ID
N0:13 from nucleotide 285 to nucleotide 590; the nucleotide sequence of SEQ ID
N0:13 from nucleotide 408 to nucleotide 590; the nucleotide sequence of the full-length protein coding sequence of clone pk855 1 deposited under accession number ATCC 98715;
or the nucleotide sequence of a mature protein coding sequence of clone pk855 1 deposited under accession number ATCC 98715. In other preferred embodiments, the polynucleotide encodes the full-length or a mature protein encoded by the cDNA
insert of clone pk855_1 deposited under accession number ATCC 98715. In further preferred embodiments, the present invention provides a polynucleotide encoding a protein comprising a fragment of the amino acid sequence of SEQ ID N0:14 having biological activity, the fragment preferably comprising eight (more preferably twenty, most preferably thirty) contiguous amino acids of SEQ ID N0:14, or a polynucleotide encoding a protein comprising a fragment of the amino acid sequence of SEQ ID N0:14 having biological activity, the fragment comprising the amino acid sequence from amino acid 46 to amino acid 55 of SEQ ID N0:14.
Other embodiments provide the gene corresponding to the cDNA sequence of SEQ
ID N0:13.
Further embodiments of the invention provide isolated polynucleotides produced according to a process selected from the group consisting of:
(a) a process comprising the steps of:
(i) preparing one or more polynucleotide probes that hybridize in 6X SSC at 65 degrees C to a nucleotide sequence selected from the group consisting of:
2 0 (aa) SEQ ID N0:13, but excluding the poly(A) tail at the 3' end of SEQ ID N0:13; and (ab) the nucleotide sequence of the cDNA insert of clone pk855 1 deposited under accession number ATCC 98715;
(ii) hybridizing said probes) to human genomic DNA in 2 5 conditions at least as stringent as 4X SSC at 50 degrees C; and (iii) isolating the DNA polynucleotides detected with the probe(s);
and (b) a process comprising the steps of:
3 0 (i) preparing one or more polynucleotide primers that hybridize in 6X SSC at 65 degrees C to a nucleotide sequence selected from the group consisting of:
(ba) SEQ ID N0:13, but excluding the poly(A) tail at the 3' end of SEQ ID N0:13; and (bb) the nucleotide sequence of the cDNA insert of clone pk855_1 deposited under accession number ATCC 98715;
(ii) hybridizing said primers) to human genomic DNA in conditions at least as stringent as 4X SSC at 50 degrees C;
(iii) amplifying human DNA sequences; and (iv) isolating the polynucleotide products of step (b)(iii).
Preferably the polynucleotide isolated according to the above process comprises a nucleotide sequence corresponding to the cDNA sequence of SEQ ID N0:13, and extending contiguously from a nucleotide sequence corresponding to the 5' end of SEQ
ID N0:13 to a nucleotide sequence corresponding to the 3' end of SEQ ID N0:13 , but excluding the poly(A) tail at the 3' end of SEQ ID N0:13. Also preferably the polynucleotide isolated according to the above process comprises a nucleotide sequence corresponding to the cDNA sequence of SEQ ID N0:13 from nucleotide 285 to nucleotide 590, and extending contiguously from a nucleotide sequence corresponding to the 5' end of said sequence of SEQ ID NO:13 from nucleotide 285 to nucleotide 590, to a nucleotide sequence corresponding to the 3' end of said sequence of SEQ ID N0:13 from nucleotide 285 to nucleotide 590. Also preferably the polynucleotide isolated according to the above process comprises a nucleotide sequence corresponding to.the cDNA sequence of SEQ ID
N0:13 from nucleotide 408 to nucleotide 590, and extending contiguously from a 2 0 nucleotide sequence corresponding to the 5' end of said sequence of SEQ ID
N0:13 from nucleotide 408 to nucleotide 590, to a nucleotide sequence corresponding to the 3' end of said sequence of SEQ ID N0:13 from nucleotide 408 to nucleotide 590.
In other embodiments, the present invention provides a composition comprising a protein, wherein said protein comprises an amino acid sequence selected from the group 2 5 consisting of:
(a) the amino acid sequence of SEQ ID N0:14;
(b) a fragment of the amino acid sequence of SEQ ID N0:14, the fragment comprising eight contiguous amino acids of SEQ ID N0:14; and (c) the amino acid sequence encoded by the cDNA insert of clone 3 0 pk855_1 deposited under accession number ATCC 98715;
the protein being substantially free from other mammalian proteins. Preferably such protein comprises the amino acid sequence of SEQ ID N0:14. In further preferred embodiments, the present invention provides a protein comprising a fragment of the amino acid sequence of SEQ ID N0:14 having biological activity, the fragment preferably comprising eight (more preferably twenty, most preferably thirty) contiguous amino acids of SEQ ID N0:14, or a protein comprising a fragment of the amino acid sequence of SEQ
ID N0:14 having biological activity, the fragment comprising the amino acid sequence from amino acid 46 to amino acid 55 of SEQ ID N0:14.
In one embodiment, the present invention provides a composition comprising an isolated polynucleotide selected from the group consisting of:
(a) a polynucleotide comprising the nucleotide sequence of SEQ ID
N0:15;
(b) a polynucleotide comprising the nucleotide sequence of SEQ ID
N0:15 from nucleotide 193 to nucleotide 2223;
(c) a polynucleotide comprising the nucleotide sequence of SEQ ID
N0:15 from nucleotide 1045 to nucleotide 2223;
(d) a polynucleotide comprising the nucleotide sequence of the full-length protein coding sequence of clone PL776_6 deposited under accession number ATCC 98715;
(e) a polynucleotide encoding the full-length protein encoded by the cDNA insert of clone PL776_6 deposited under accession number ATCC 98715;
(f) a polynucleotide comprising the nucleotide sequence of a mature protein coding sequence of clone PL776 6 deposited under accession number 2 0 ATCC 98715;
(g) a polynucleotide encoding a mature protein encoded by the cDNA
insert of clone PL776 6 deposited under accession number ATCC 98715;
(h) a polynucleotide encoding a protein comprising the amino acid sequence of SEQ ID N0:16;
2 5 (i) a polynucleotide encoding a protein comprising a fragment of the amino acid sequence of SEQ ID N0:16 having biological activity, the fragment comprising eight contiguous amino acids of SEQ ID N0:16;
(j) a polynucleotide which is an allelic variant of a polynucleotide of (a)-(g) above;
3 0 (k) a polynucleotide which encodes a species homologue of the protein of (h) or (i) above ;
(1) a polynucleotide that hybridizes under stringent conditions to any one of the polynucleotides specified in (a)-(i); and (m) a polynucleotide that hybridizes under stringent conditions to any one of the polynucleotides specified in (a)-(i) and that has a length that is at least 25% of the length of SEQ ID N0:15.
Preferably, such polynucleotide comprises the nucleotide sequence of SEQ ID
N0:15 from nucleotide 193 to nucleotide 2223; the nucleotide sequence of SEQ
ID NO:15 from nucleotide 1045 to nucleotide 2223; the nucleotide sequence of the full-length protein coding sequence of clone PL776_6 deposited under accession number ATCC
98715; or the nucleotide sequence of a mature protein coding sequence of clone PL776_6 deposited under accession number ATCC 98715. In other preferred embodiments, the polynucleotide encodes the full-length or a mature protein encoded by the cDNA
insert of clone PL776_6 deposited under accession number ATCC 98715. In further preferred embodiments, the present invention provides a polynucleotide encoding a protein comprising a fragment of the amino acid sequence of SEQ ID N0:16 having biological activity, the fragment preferably comprising eight (more preferably twenty, most preferably thirty) contiguous amino acids of SEQ ID N0:16, or a polynucleotide encoding a protein comprising a fragment of the amino acid sequence of SEQ ID N0:16 having biological activity, the fragment comprising the amino acid sequence from amino acid 333 to amino acid 342 of SEQ ID N0:16.
Other embodiments provide the gene corresponding to the cDNA sequence of SEQ
2 0 ID N0:15.
Further embodiments of the invention provide isolated polynucleotides produced according to a process selected from the group consisting of:
(a) a process comprising the steps of:
(i) preparing one or more polynucleotide probes that hybridize 2 5 in 6X SSC at 65 degrees C to a nucleotide sequence selected from the group consisting of:
(aa) SEQ ID N0:15; and (ab) the nucleotide sequence of the cDNA insert of clone PL776_6 deposited under accession number ATCC 98715;
3 0 (ii) hybridizing said probes) to human genomic DNA in conditions at least as stringent as 4X SSC at 50 degrees C; and (iii) isolating the DNA polynucleotides detected with the probe(s);
and (b) a process comprising the steps of:
(i) preparing one or more polynucleotide primers that hybridize in 6X SSC at 65 degrees C to a nucleotide sequence selected from the group consisting of:
(ba) SEQ ID N0:15; and (bb) the nucleotide sequence of the cDNA insert of clone PL776_6 deposited under accession number ATCC 98715;
(ii) hybridizing said primers) to human genomic DNA in conditions at least as stringent as 4X SSC at 50 degrees C;
(iii) amplifying human DNA sequences; and (iv) isolating the polynucleotide products of step (b)(iii).
Preferably the polynucleotide isolated according to the above process comprises a nucleotide sequence corresponding to the cDNA sequence of SEQ ID N0:15, and extending contiguously from a nucleotide sequence corresponding to the 5' end of SEQ
ID N0:15 to a nucleotide sequence corresponding to the 3' end of SEQ ID N0:15 . Also preferably the polynucleotide isolated according to the above process comprises a nucleotide sequence corresponding to the cDNA sequence of SEQ ID N0:15 from nucleotide 193 to nucleotide 2223, and extending contiguously from a nucleotide sequence corresponding to the 5' end of said sequence of SEQ ID N0:15 from nucleotide 193 to 2 0 nucleotide 2223, to a nucleotide sequence corresponding to the 3' end of said sequence of SEQ ID N0:15 from nucleotide 193 to nucleotide 2223. Also preferably the polynucleotide isolated according to the above process comprises a nucleotide sequence corresponding to the cDNA sequence of SEQ ID N0:15 from nucleotide 1045 to nucleotide 2223, and extending contiguously from a nucleotide sequence corresponding to the 5' end of said 2 5 sequence of SEQ ID N0:15 from nucleotide 1045 to nucleotide 2223, to a nucleotide sequence corresponding to the 3' end of said sequence of SEQ ID N0:15 from nucleotide 1045 to nucleotide 2223.
In other embodiments, the present invention provides a composition comprising a protein, wherein said protein comprises an amino acid sequence selected from the group 3 0 consisting of:
(a) the amino acid sequence of SEQ ID N0:16;
(b) a fragment of the amino acid sequence of SEQ ID N0:16, the fragment comprising eight contiguous amino acids of SEQ ID N0:16; and (c) the amino acid sequence encoded by the cDNA insert of clone PL776_6 deposited under accession number ATCC 98715;
the protein being substantially free from other mammalian proteins. Preferably such protein comprises the amino acid sequence of SEQ ID N0:16. In further preferred embodiments, the present invention provides a protein comprising a fragment of the amino acid sequence of SEQ ID N0:16 having biological activity, the fragment preferably comprising eight (more preferably twenty, most preferably thirty) contiguous amino acids of SEQ ID N0:16, or a protein comprising a fragment of the amino acid sequence of SEQ
ID N0:16 having biological activity, the fragment comprising the amino acid sequence from amino acid 333 to amino acid 342 of SEQ ID N0:16.
In one embodiment, the present invention provides a composition comprising an isolated polynucleotide selected from the group consisting of:
(a) a polynucleotide comprising the nucleotide sequence of SEQ ID
N0:17;
(b) a polynucleotide comprising the nucleotide sequence of SEQ ID
N0:17 from nucleotide 198 to nucleotide 1121;
(c) a polynucleotide comprising the nucleotide sequence of SEQ ID
N0:17 from nucleotide 381 to nucleotide 1121;
(d) a polynucleotide comprising the nucleotide sequence of the full-2 0 length protein coding sequence of clone pm4_13 deposited under accession number ATCC 98715;
(e) a polynucleotide encoding the full-length protein encoded by the cDNA insert of clone pm4_13 deposited under accession number ATCC 98715;
(f) a polynucleotide comprising the nucleotide sequence of a mature 2 5 protein coding sequence of clone pm4_13 deposited under accession number ATCC 98715;
(g) a polynucleotide encoding a mature protein encoded by the cDNA
insert of clone pm4_13 deposited under accession number ATCC 98715;
(h) a polynucleotide encoding a protein comprising the amino acid 3 0 sequence of SEQ ID N0:18;
(i) a polynucleotide encoding a protein comprising a fragment of the amino acid sequence of SEQ ID N0:18 having biological activity, the fragment comprising eight contiguous amino acids of SEQ ID N0:18;

(j) a polynucleotide which is an allelic variant of a polynucleotide of (a)-(g) above;
(k) a polynucleotide which encodes a species homologue of the protein of (h) or (i) above ;
(1) a polynucleotide that hybridizes under stringent conditions to any one of the polynucleotides specified in (a)-(i); and (m) a polynucleotide that hybridizes under stringent conditions to any one of the polynucleotides specified in (a)-(i) and that has a length that is at least 25% of the length of SEQ ID N0:17.
Preferably, such polynucleotide comprises the nucleotide sequence of SEQ ID
N0:17 from nucleotide 198 to nucleotide 1121; the nucleotide sequence of SEQ
ID N0:17 from nucleotide 381 to nucleotide 1121; the nucleotide sequence of the full-length protein coding sequence of clone pm4_13 deposited under accession number ATCC 98715;
or the nucleotide sequence of a mature protein coding sequence of clone pm4_13 deposited under accession number ATCC 98715. In other preferred embodiments, the polynucleotide encodes the full-length or a mature protein encoded by the cDNA
insert of clone pm4_13 deposited under accession number ATCC 98715. In further preferred embodiments, the present invention provides a polynucleotide encoding a protein comprising a fragment of the amino acid sequence of SEQ ID N0:18 having biological 2 0 activity, the fragment preferably comprising eight (more preferably twenty, most preferably thirty} contiguous amino ands of SEQ ID N0:18, or a polynucleotide encoding a protein comprising a fragment of the amino acid sequence of SEQ ID N0:18 having biological activity, the fragment comprising the amino acid sequence from amino acid 149 to amino acid 158 of SEQ ID N0:18.
2 5 Other embodiments provide the gene corresponding to the cDNA sequence of SEQ
ID N0:17.
Further embodiments of the invention provide isolated polynucleotides produced according to a process selected from the group consisting of:
(a) a process comprising the steps of:
3 0 (i) preparing one or more polynucleotide probes that hybridize in 6X SSC at 65 degrees C to a nucleotide sequence selected from the group consisting of:
(aa) SEQ ID N0:17, but excluding the poly(A) tail at the 3' end of SEQ ID N0:17; and (ab) the nucleotide sequence of the cDNA insert of clone pm4_13 deposited under accession number ATCC 98715;
(ii) hybridizing said probes) to human genomic DNA in conditions at least as stringent as 4X SSC at 50 degrees C; and (iii) isolating the DNA polynucleotides detected with the probe(s);
and (b) a process comprising the steps of:
(i) preparing one or more polynucleotide primers that hybridize in 6X SSC at 65 degrees C to a nucleotide sequence selected from the group consisting of:
(ba) SEQ ID N0:17, but excluding the poly(A) tail at the 3' end of SEQ ID N0:17; and (bb) the nucleotide sequence of the cDNA insert of clone ~ 5 pm4_13 deposited under accession number ATCC 98715;
(ii) hybridizing said primers) to human genomic DNA in conditions at least as stringent as 4X SSC at 50 degrees C;
(iii) amplifying human DNA sequences; and (iv) isolating the polynucleotide products of step (b)(iii).
2 0 Preferably the polynucleotide isolated according to the above process comprises a nucleotide sequence corresponding to the cDNA sequence of SEQ ID N0:17, and extending contiguously from a nucleotide sequence corresponding to the 5' end of SEQ
ID N0:17 to a nucleotide sequence corresponding to the 3' end of SEQ ID N0:17 , but excluding the poly(A) tail at the 3' end of SEQ ID N0:17. Also preferably the 2 5 polynucleotide isolated according to the above process comprises a nucleotide sequence corresponding to the cDNA sequence of SEQ ID N0:17 from nucleotide 198 to nucleotide 1121, and extending contiguously from a nucleotide sequence corresponding to the 5' end of said sequence of SEQ ID N0:17 from nucleotide 198 to nucleotide 1121, to a nucleotide sequence corresponding to the 3' end of said sequence of SEQ ID N0:17 from nucleotide 3 0 198 to nucleotide 1121. Also preferably the polynucleotide isolated according to the above process comprises a nucleotide sequence corresponding to the cDNA sequence of SEQ ID
N0:17 from nucleotide 381 to nucleotide 1121, and extending contiguously from a nucleotide sequence corresponding to the 5' end of said sequence of SEQ ID
N0:17 from nucleotide 381 to nucleotide 1121, to a nucleotide sequence corresponding to the 3' end of said sequence of SEQ ID N0:17 from nucleotide 381 to nucleotide 1121.
In other embodiments, the present invention provides a composition comprising a protein, wherein said protein comprises an amino acid sequence selected from the group consisting of:
(a) the amino acid sequence of SEQ ID N0:18;
(b) a fragment of the amino acid sequence of SEQ ID N0:18, the fragment comprising eight contiguous amino acids of SEQ ID NO:18; and (c) the amino acid sequence encoded by the cDNA insert of clone pm4_13 deposited under accession number ATCC 98715;
the protein being substantially free from other mammalian proteins. Preferably such protein comprises the amino acid sequence of SEQ ID N0:18. In further preferred embodiments, the present invention provides a protein comprising a fragment of the amino acid sequence of SEQ ID NO:I8 having biological activity, the fragment preferably comprising eight (more preferably twenty, most preferably thirty) contiguous amino acids of SEQ ID N0:18, or a protein comprising a fragment of the amino acid sequence of SEQ
ID N0:18 having biological activity, the fragment comprising the amino acid sequence from amino acid 149 to amino acid 158 of SEQ ID N0:18.
In one embodiment, the present invention provides a composition comprising an 2 0 isolated polynucleotide selected from the group consisting of:
(a) a polynucleotide comprising the nucleotide sequence of SEQ ID
N0:19;
(b) a polynucleotide comprising the nucleotide sequence of SEQ ID
N0:19 from nucleotide 19 to nucleotide 1953;
2 5 (c) a polynucleotide comprising the nucleotide sequence of the full-length protein coding sequence of clone pt326_4 deposited under accession number ATCC 98715;
(d) a polynucleotide encoding the full-length protein encoded by the cDNA insert of clone pt326 4 deposited under accession number ATCC 98715;
3 0 (e) a polynucleotide comprising the nucleotide sequence of a mature protein coding sequence of clone pt326 4 deposited under accession number ATCC 98715;
(f) a polynucleotide encoding a mature protein encoded by the cDNA
insert of clone pt326 4 deposited under accession number ATCC 98715;

(g) a polynucleotide encoding a protein comprising the amino acid sequence of SEQ ID N0:20;
(h) a polynucleotide encoding a protein comprising a fragment of the amino acid sequence of SEQ ID N0:20 having biological activity, the fragment comprising eight contiguous amino acids of SEQ ID N0:20;
(i) a polynucleotide which is an allelic variant of a polynucleotide of (a)-(f) above;
(j) a polynucleotide which encodes a species homologue of the protein of (g) or (h) above ;
(k) a polynucleotide that hybridizes under stringent conditions to any one of the polynucleotides specified in (a)-(h); and (1) a polynucleotide that hybridizes under stringent conditions to any one of the polynucleotides specified in (a~(h) and that has a length that is at least 25% of the length of SEQ ID N0:19.
Preferably, such polynucleotide comprises the nucleotide sequence of SEQ ID
N0:19 from nucleotide 19 to nucleotide 1953; the nucleotide sequence of the full-length protein coding sequence of clone pt326_4 deposited under accession number ATCC
98715;
or the nucleotide sequence of a mature protein coding sequence of clone pt326_4 deposited under accession number ATCC 98715. In other preferred embodiments, the 2 0 polynucleotide encodes the full-length or a mature protein encoded by the cDNA insert of clone pt326 4 deposited under accession number ATCC 98715. In further preferred embodiments, the present invention provides a polynucleotide encoding a protein comprising a fragment of the amino acid sequence of SEQ ID N0:20 having biological activity, the fragment preferably comprising eight (more preferably twenty, most 2 5 preferably thirty) contiguous amino acids of SEQ ID N0:20, or a polynucleotide encoding a protein comprising a fragment of the amino acid sequence of SEQ ID N0:20 having biological activity, the fragment comprising the amino acid sequence from amino acid 317 to amino acid 326 of SEQ ID N0:20.
Other embodiments provide the gene corresponding to the cDNA sequence of SEQ
3 0 ID N0:19.
Further embodiments of the invention provide isolated polynucleotides produced according to a process selected from the group consisting of:
(a) a process comprising the steps of:

(i) preparing one or more polynucleotide probes that hybridize in 6X SSC at 65 degrees C to a nucleotide sequence selected from the group consisting of:
(aa) SEQ ID N0:19, but excluding the poly(A) tail at the 3' end of SEQ ID N0:19; and {ab) the nucleotide sequence of the cDNA insert of clone pt326_4 deposited under accession number ATCC 98715;
(ii) hybridizing said probes) to human eenomic DNA in conditions at least as stringent as 9X SSC at 50 degrees C; and (iii) isolating the DNA polynucleotides detected with the probe(s);
and (b) a process comprising the steps of:
{i) preparing one or more polynucleotide primers that hybridize in 6X SSC at 65 degrees C to a nucleotide sequence selected from the group consisting of:
(ba) SEQ ID N0:19, but excluding the poly(A) tail at the 3' end of SEQ ID N0:19; and (bb) the nucleotide sequence of the cDNA insert of done 2 0 pt326 4 deposited under accession number ATCC 98715;
(ii) hybridizing said primers) to human genomic DNA in conditions at least as stringent as 4X SSC at 50 degrees C;
(iii) amplifying human DNA sequences; and (iv) isolating the polynucleotide products of step (b)(iii).
2 5 Preferably the polynucleotide isolated according to the above process comprises a nucleotide sequence corresponding to the cDNA sequence of SEQ ID N0:19, and extending contiguously from a nucleotide sequence corresponding to the 5' end of SEQ
ID N0:19 to a nucleotide sequence corresponding to the 3' end of SEQ ID N0:19 , but excluding the poly(A) tail at the 3' end of SEQ ID N0:19. Also preferably the 3 0 polynucleotide isolated according to the above process comprises a nucleotide sequence corresponding to the cDNA sequence of SEQ ID NO:I9 from nucleotide 19 to nucleotide 1953, and extending contiguously from a nucleotide sequence corresponding to the 5' end of said sequence of SEQ ID N0:19 from nucleotide 19 to nucleotide 1953, to a nucleotide sequence corresponding to the 3' end of said sequence of SEQ ID N0:19 from nucleotide 19 to nucleotide 1953.
In other embodiments, the present invention provides a composition comprising a protein, wherein said protein comprises an amino acid sequence selected from the group consisting of:
(a) the amino acid sequence of SEQ ID N0:20;
(b) a fragment of the amino acid sequence of SEQ ID N0:20, the fragment comprising eight contiguous amino acids of SEQ ID N0:20; and (c) the amino acid sequence encoded by the cDNA insert of clone pt326 4 deposited under accession number ATCC 98715;
the protein being substantially free from other mammalian proteins. Preferably such protein comprises the amino acid sequence of SEQ ID N0:20. In further preferred embodiments, the present invention provides a protein comprising a fragment of the amino and sequence of SEQ ID N0:20 having biological activity, the fragment preferably comprising eight (more preferably twenty, most preferably thirty) contiguous amino acids of SEQ ID N0:20, or a protein comprising a fragment of the amino acid sequence of SEQ
ID N0:20 having biological activity, the fragment comprising the amino acid sequence from amino acid 317 to amino acid 326 of SEQ ID N0:20.
In certain preferred embodiments, the polynucleotide is operably linked to an 2 0 expression control sequence. The invention also provides a host cell, including bacterial, yeast, insect and mammalian cells, transformed with such polynucleotide compositions.
Also provided by the present invention are organisms that have enhanced, reduced, or modified expression of the genes) corresponding to the polynucleotide sequences disclosed herein.
2 5 Processes are also provided for producing a protein, which comprise:
(a) growing a culture of the host cell transformed with such polynucleotide compositions in a suitable culture medium; and (b) purifying the protein from the culture.
The protein produced according to such methods is also provided by the present 3 0 invention.
Protein compositions of the present invention may further comprise a pharmaceutically acceptable carrier. Compositions comprising an antibody which specifically reacts with such protein are also provided by the present invention.

Methods are also provided for preventing, treating or ameliorating a medical condition which comprises administering to a mammalian subject a therapeutically effective amount of a composition comprising a protein of the present invention and a pharmaceutically acceptable Garner.
BRIEF DESCRIPTION OF THE DRAWINGS
Figures lA and 1B are schematic representations of the pED6 and pNOTs vectors, respectively, used for deposit of clones disclosed herein.
DETAILED DESCRIIrTION
ISOLATED PROTEINS AND POLYNUCLEOTIDES
Nucleotide and amino acid sequences, as presently determined, are reported below for each clone and protein disclosed in the present application. The nucleotide sequence of each clone can readily be determined by sequencing of the deposited clone in accordance with known methods. The predicted amino acid sequence (both full-length and mature forms) can then be determined from such nucleotide sequence. The amino acid sequence of the protein encoded by a particular clone can also be determined by expression of the clone in a suitable host cell, collecting the protein and determining its sequence. For each disclosed protein applicants have identified what they have 2 0 determined to be the reading frame best identifiable with sequence information available at the time of filing.
As used herein a "secreted" protein is one which, when expressed in a suitable host cell, is transported across or through a membrane, including transport as a result of signal sequences in its amino acaid sequence. "Secreted" proteins include without limitation 2 5 proteins secreted wholly (e.g., soluble proteins) or partially (e.g. , receptors) from the cell in which they are expressed. "Secreted" proteins also include without limitation proteins which are transported across the membrane of the endoplasmic reticulurn.
Clone "cc359 4"
3 0 A polynucleotide of the present invention has been identified as clone "cc359 4".
cc359 4 was isolated from a human adult brain cDNA library using methods which are selective for cDNAs encoding secreted proteins (see U.S. Pat. No. 5,536,637), or was identified as encoding a secreted or transmembrane protein on the basis of computer analysis of the amino acid sequence of the encoded protein. cc359 4 is a full-length clone, including the entire coding sequence of a secreted protein (also referred to herein as "cc359 4 protein").
The nucleotide sequence of cc359_4 as presently determined is reported in SEQ
ID
NO:1, and includes a poly(A) tail. What applicants presently believe to be the proper reading frame and the predicted amino acid sequence of the cc359 4 protein corresponding to the foregoing nucleotide sequence is reported in SEQ ID N0:2.
Amino acids 29 to 41 of SEQ ID N0:2 are a predicted leader/signal sequence, with the predicted mature amino acid sequence beginning at amino acid 42. Due to the hydrophobic nature of the predicted leader/signal sequence, it is likely to act as a transmembrane domain should the predicted leader/signal sequence not be separated from the remainder of the cc359 4 protein.
The EcoRI/NotI restriction fragment obtainable from the deposit containing clone cc359 4 should be approximately 2200 bp.
The nucleotide sequence disclosed herein for cc359 4 was searched against the GenBank and GeneSeq nucleotide sequence databases using BLASTN/BLASTX and FASTA search protocols. cc359 4 demonstrated at least some similarity with sequences identified as H23117 (ym51g12.s1 Homo sapiens cDNA clone 51970 3'). Based upon sequence similarity, cc359 4 proteins and each similar protein or peptide may share at least some activity. The cc359 4 protein sequence contains four chaoptin domains (at 2 0 amino acids 94-115,118-139,142-163, and 261-282 of SEQ ID N0:2). Chaoptin is a cell-surface glycoprotein required for Drosophiln photoreceptor cell morphogenesis.
Chaoptin is largely composed of 41 potentially amphipathic repeats. Chaoptin repeats that have been reported in both yeast and human proteins are remarkably similar, suggesting their general importance as a structural and/or functional motif. Additionally, the cc359_4 2 5 protein was found to contain a leucine zipper motif. The TopPredlT
computer program predicts five potential transmembrane domains within the cc359_4 protein sequence, centered around amino acids 20, 410, 490, 530, and 590 of SEQ ID N0:2, respectively.
Clone "ct547 2"
3 0 A polynucleotide of the present invention has been identified as clone "ct547 2".
ct547_2 was isolated from a human adult brain cDNA library using methods which are selective for cDNAs encoding secreted proteins (see U.S. Pat. No. 5,536,637), or was identified as encoding a secreted or transmembrane protein on the basis of computer analysis of the amino acid sequence of the encoded protein. ct547_2 is a full-length clone, including the entire coding sequence of a secreted protein (also referred to herein as "ct547 2 protein").
The nucleotide sequence of ct547_2 as presently determined is reported in SEQ
ID
N0:3, and includes a poly(A) tail. What applicants presently believe to be the proper reading frame and the predicted amino acid sequence of the ct547_2 protein corresponding to the foregoing nucleotide sequence is reported in SEQ ID N0:4.
Amino acids 44 to 56 of SEQ ID N0:4 are a predicted leader/signal sequence, with the predicted mature amino acid sequence beginning at amino acid 57. Due to the hydrophobic nature of the predicted Ieader/signal sequence, it is likely to act as a transmembrane domain should the predicted leader/signal sequence not be separated from the remainder of the ct547 2 protein.
The EcoRI/NotI restriction fragment obtainable from the deposit containing clone ct547 2 should be approximately 1600 bp.
The nucleotide sequence disclosed herein for ct547 2 was searched against the 25 GenBank and GeneSeq nucleotide sequence databases using BLASTN/BLASTX and FASTA search protocols. ct547 2 demonstrated at least some similarity with sequences identified as AA428546 (zw47d03.r1 Soares total fetus Nb2HF8 9w Homo sapiens cDNA
clone 773189 5'}, 820032 (yg31g04.r1 Homo sapiens cDNA clone 34069 5'), 827350 (yh53c12.r1 Homo Sapiens cDNA clone 133462 5'), and 866971 (yi29e12.r1 Homo sapiens 2 0 cDNA clone 140686 5'). Based upon sequence similarity, ct547_2 proteins and each similar protein or peptide may share at least some activity.
Clone "en553 1"
A polynucleotide of the present invention has been identified as clone "en553 1".
2 5 en553_1 was isolated from a human fetal brain cDNA Library and was identified as encoding a secreted or transmembrane protein on the basis of computer analysis of the amino acid sequence of the encoded protein. en553_1 is a full-length clone, including the entire coding sequence of a secreted protein (also referred to herein as "en553_1 protein").
The nucleotide sequence of en553_1 as presently determined is reported in SEQ
3 0 ID N0:5, and includes a poly(A) tail. What applicants presently believe to be the proper reading frame and the predicted amino acid sequence of the en553_1 protein corresponding to the foregoing nucleotide sequence is reported in SEQ ID N0:6.
Amino acids 17 to 29 of SEQ ID N0:6 are a predicted leader/signal sequence, with the predicted mature amino acid sequence beginning at amino acid 30. Due to the hydrophobic nature of the predicted leader/signal sequence, it is likely to act as a transmembrane domain should the predicted leader/signal sequence not be separated from the remainder of the en553_1 protein. Another potential en553_1 reading frame and predicted amino acid sequence is encoded by basepairs 1334 to 1597 of SEQ ID N0:5 and is reported in SEQ ID
N0:31 The EcoRI/NotI restriction fragment obtainable from the deposit containing clone en553_1 should be approximately 2675 bp.
The nucleotide sequence disclosed herein for en553_1 was searched against the GenBank and GeneSeq nucleotide sequence databases using BLASTN/BLASTX and FASTA search protocols. en553_1 demonstrated at least some similarity with sequences identified as No hits were found in the databases. The nucleotide sequence of en553_1 indicates that it may contain one or more copies of the following repetitive elements: Alu, MIIt.
Clone "nn296 2"
A polynucleotide of the present invention has been identified as clone "nn296 2".
nn296_2 was isolated from a human fetal kidney (293 cell line) cDNA library using methods which are selective for cDNAs encoding secreted proteins (see U.S.
Pat. No.
5,536,637), or was identified as encoding a secreted or transmembrane protein on the basis 2 0 of computer analysis of the amino acid sequence of the encoded protein.
nn296 2 is a full-length clone, including the entire coding sequence of a secreted protein (also referred to herein as "nn296_2 protein").
The nucleotide sequence of nn296 2 as presently determined is reported in SEQ
ID N0:7, and includes a poly(A) tail. What applicants presently believe to be the proper 2 5 reading frame and the predicted amino acid sequence of the nn296 2 protein corresponding to the foregoing nucleotide sequence is reported in SEQ ID N0:8.
The EcoRI/NotI restriction fragment obtainable from the deposit containing clone nn296_2 should be approximately 4500 bp.
The nucleotide sequence disclosed herein for nn296_2 was searched against the 3 0 GenBank and GeneSeq nucleotide sequence databases using BLASTN/BLASTX and FASTA search protocols. nn296_2 demonstrated at least some similarity with sequences identified as AA054538 (zk83e02.s1 Soares pregnant uterus NbHPU Homo sapiens cDNA
clone 489434 3'), AA175407 (ms80c06.r1 Soares mouse 3NbMS Mus musculus cDNA
clone 617866 5' similar to WP R08C7.2 CE07425), AA510586 (vg33c05.r1 Soares mouse mammary gland NbMMG Mus musculus cDNA clone 863144 5' similar to WP:R08C7.2 CE07425), C18077 (Human placenta cDNA 5'-end GEN-557B03}, T24416 (Human gene signature HUMGS06449), and W74238 (zd02f09.r1 Pancreatic Islet Homo sapiens cDNA
clone 339497 5'). The predicted amino acid sequence disclosed herein for nn296_2 was searched against the GenPept and GeneSeq amino acid sequence databases using the BLASTX search protocol. The predicted nn296 2 protein demonstrated at least some similarity to sequences identified as U61953 (cosmid R08C7 [Caenorhabditis elegansj).
Based upon sequence similarity, nn296 2 proteins and each similar protein or peptide may share at least some activity. The TopPredII computer program predicts two potential transmembrane domains within the nn296 2 protein sequence, one centered around amino acid 50 and another around amino acid 340 of SEQ ID N0:8. The nucleotide sequence of nn296 2 indicates that it may contain at least one repetitive element.
Clone "n 2q 7 13"
A polynucleotide of the present invention has been identified as clone "nq27 13".
nq27 13 was isolated from a human adult blood (erythroleukemia TF-1) cDNA
library using methods which are selective for cDNAs encoding secreted proteins (see U.S. Pat.
No. 5,536,637), or was identified as encoding a secreted or transmembrane protein on the basis of computer analysis of the amino acid sequence of the encoded protein.
nq27 13 2 0 is a full-length clone, including the entire coding sequence of a secreted protein (also referred to herein as "nq27 13 protein").
The nucleotide sequence of nq27 13 as presently determined is reported in SEQ
ID N0:9, and includes a poly(A) tail. What applicants presently believe to be the proper reading frame and the predicted amino acid sequence of the nq27 13 protein 2 5 corresponding to the foregoing nucleotide sequence is reported in SEQ ID
NO:10.
Another potential nq27 13 reading frame and predicted amino acid sequence is encoded by basepairs 371 to 616 of SEQ ID N0:9 and is reported in SEQ ID N0:32.
The EcoRI/NotI restriction fragment obtainable from the deposit containing clone nq27 13 should be approximately 900 bp.
3 0 The nucleotide sequence disclosed herein for nq27_23 was searched against the GenBank and GeneSeq nucleotide sequence databases using BLASTN/BLASTX and FASTA search protocols. nq27 13 demonstrated at least some similarity with sequences identified as AA015599 (ze20h10.s1 Soares fetal heart NbHHI9W Homo sapiens cDNA
clone 359587 3'), AA465I12 (aa32c05.r1 NCI CGAP_GCBl Homo Sapiens cDNA clone IMAGE:814952 5'), AA628397 (af26a12.sI Soares total fetus Nb2HF8 9w Homo sapiens cDNA clone 1032766 3'), AA628438 (af26e12.s1 Soares total fetus Nb2HF8 9w Homo Sapiens cDNA done 1032814 3'), D79280 (Human aorta cDNA 5'-end GEN-213H05), and H81836 (ys68g11.r1 Homo sapiens cDNA clone 220004 5'). Based upon sequence similarity, nq27 13 proteins and each similar protein or peptide may share at least some activity. The TopPredII computer program predicts a potential transmembrane domain at the N-terminus of the nq27 13 protein sequence, centered around amino arid 13 of SEQ
ID NO:10.
nq27 13 protein was expressed in a COS cell expression system, and an expressed protein band of approximately 11 kDa was detected in membrane fractions using SDS
polyacrylamide gel electrophoresis.
Clone "pk65 4"
A polynucleotide of the present invention has been identified as clone "pk65 4".
pk65 4 was isolated from a human fetal kidney (293 cell line) cDNA library using methods which are selective for cDNAs encoding secreted proteins (see U.S.
Pat. No.
5,536,637), or was identified as encoding a secreted or transmembrane protein on the basis of computer analysis of the amino acid sequence of the encoded protein. pk65 4 is a full length clone, including the entire coding sequence of a secreted protein (also referred to 2 0 herein as "pk65 4 protein").
The nucleotide sequence of pk65 4 as presently determined is reported in SEQ
ID
N0:11, and includes a poly(A) tail. What applicants presently believe to be the proper reading frame and the predicted amino acid sequence of the pk65 4 protein corresponding to the foregoing nucleotide sequence is reported in SEQ ID
N0:12.
2 5 The EcoRI/NotI restriction fragment obtainable from the deposit containing clone pk65 4 should be approximately 1500 bp.
The nucleotide sequence disclosed herein for pk65 4 was searched against the GenBank and GeneSeq nucleotide sequence databases using BLASTN/BLASTX and FASTA search protocols. pk65 4 demonstrated at least some similarity with sequences 3 0 identified as AA309041 (EST179822 Colon carcinoma (Caco-2) cell line I
Homo Sapiens cDNA 5' end), AA316187 (EST187903 HCC cell line (matastasis to liver in mouse) II Homo sapiens cDNA 5' end), AA488843 (aa55a10.s1 NCI CGAP_GCB1 Homo sapiens cDNA
clone IMAGE:824826 3'), AF022811 (Mus musculus cornichon mItNA, complete cds), D16980 (Human HepG2 partial cDNA, clone hmd2a04m5), N44081 (yy32a02.r1 Homo Sapiens cDNA clone 272906 5' similar to PIR A56724 A56724 cni protein - fruit fly). The predicted amino acid sequence disclosed herein for pk65_4 was searched against the GenPept and GeneSeq amino acid sequence databases using the B1.ASTX search protocol.
The predicted pk65 4 protein demonstrated at Ieast some similarity to sequences identified as AB006191 (cornichon-like protein [Mus musculus]), AF022811 (cornichon (Mus musculus]), and U28069 (cni gene product (Drosophila melanogasterJ).
Based upon sequence similarity, pk65_4 proteins and each similar protein or peptide may share at least some activity. The TopPredII computer program predicts three potential transmembrane domains within the pk65 4 protein sequence, centered around amino acids 16, 67, and 133 of SEQ ID N0:12, respectively.
pk65 4 protein was expressed in a COS cell expression system, and an expressed protein band of approximately 15 kDa was detected in membrane fractions using SDS
polyacrylamide gel electrophoresis.
Clone "pk85 1"
A polynucleotide of the present invention has been identified as clone "pk855_1".
pk855_1 was isolated from a human fetal kidney (293 cell Line) cDNA library using methods which are selective for cDNAs encoding secreted proteins (see U.S.
Pat. No.
5,536,637), or was identified as encoding a secreted or transmembrane protein on the basis 2 0 of computer analysis of the amino acid sequence of the encoded protein.
pk855_1 is a full-length clone, including the entire coding sequence of a secreted protein (also referred to herein as "pk855_1 protein").
The nucleotide sequence of pk855_1 as presently determined is reported in SEQ
ID N0:13, and includes a poly(A) tail. What applicants presently believe to be the proper 2 5 reading frame and the predicted amino acid sequence of the pk855_1 protein corresponding to the foregoing nucleotide sequence is reported in SEQ ID
N0:14. Amino acids 29 to 41 of SEQ ID N0:14 are a predicted leader/signal sequence, with the predicted mature amino acid sequence beginning at amino acid 42. Due to the hydrophobic nature of the predicted leader/signal sequence, it is likely to act as a transmembrane domain 3 0 should the predicted leader/signal sequence not be separated from the remainder of the pk855_1 protein.
The EcoRI/NotI restriction fragment obtainable from the deposit containing clone pk855_1 should be approximately 1400 bp.

The nucleotide sequence disclosed herein for pk855_1 was searched against the GenBank and GeneSeq nucleotide sequence databases using BLASTN/BLASTX and FASTA search protocols. pk855_1 demonstrated at least some similarity with sequences identified as AA236320 (zr53h03.r1 Soares NI~iMPu S1 Homo Sapiens cDNA clone 667157 5'), AA491296 (aa53d03.s1 NCI CGAP_GCBl Homo Sapiens cDNA clone ILVIAGE:824645 3'), and W76249 (zd66e06.r1 Soares fetal heart NbHHI9W Homo sapiens cDNA clone 345634 5'). Based upon sequence similarity, pk855_1 proteins and each similar protein or peptide may share at least some activity. The TopPredlI
computer program predicts an additional potential transmembrane domain within the pk855_1 protein sequence centered around amino acid 78 of SEQ ID N0:14. The nucleotide sequence of pk855_1 indicates that it may contain some repeat elements (including some short stretches of poly(T~).
Clone "PL776 6"
A polynucleotide of the present invention has been identified as clone "PL776 6".
PL776_6 was isolated from a human fetal kidney (293 cell line) cDNA library using methods which are selective for cDNAs encoding secreted proteins (see U.S.
Pat. No.
5,536,637), or was identified as encoding a secreted or transmembrane protein on the basis of computer analysis of the amino acid sequence of the encoded protein.
PL776_6 is a full-2 0 length clone, including the entire coding sequence of a secreted protein (also referred to herein as "PL776_6 protein").
The nucleotide sequence of PL776_6 as presently determined is reported in SEQ
ID N0:15, and includes a poly(A) tail. What applicants presently believe to be the proper reading frame and the predicted amino acid sequence of the PL776 6 protein 2 5 corresponding to the foregoing nucleotide sequence is reported in SEQ ID
N0:16. Amino acids 272 to 284 of SEQ ID N0:16 are a predicted leader/signal sequence, with the predicted mature amino acid sequence beginning at amino acid 285. Due to the hydrophobic nature of the predicted leader/signal sequence, it is likely to act as a transmembrane domain should the predicted leader/signal sequence not be separated 3 0 from the remainder of the PL776_6 protein.
The EcoRI/NotI restriction fragment obtainable from the deposit containing clone PL776_6 should be approximately 3600 bp.
The nucleotide sequence disclosed herein for PL776_6 was searched against the GenBank and GeneSeq nucleotide sequence databases using BLASTN/BLASTX and FASTA search protocols. PL776 6 demonstrated at least some similarity with sequences identified as AA009770 (zi04c06.s1 Soares fetal liver spleen 1NFLS Sl Homo sapiens cDNA clone 429802 3'), AA056343 (z166f04.s1 Stratagene colon (#937204) Homo Sapiens cDNA clone 509599 3'}, AA076433 (zm19g07.s1 Stratagene pancreas (#937208) Homo sapiens cDNA clone 526140 3'), AA076517 (zm19g07.r1 Stratagene pancreas (#937208) Homo Sapiens cDNA clone 526140 5'), M93661 (Rat notch 2 mRNA), and 836803 (CBS-Homo Sapiens cDNA clone CBS-216 5' end). Based upon sequence similarity, PL776_6 proteins and each similar protein or peptide may share at least some activity.
Clone " m A polynucleotide of the present invention has been identified as clone "pm4_13".
pm4_I3 was isolated from a human fetal kidney (293 cell line) cDNA library using methods which are selective for cDNAs encoding secreted proteins (see U.S.
Pat. No.
5,536,637), or was identified as encoding a secreted or transmembrane protein on the basis of computer analysis of the amino acid sequence of the encoded protein. pm4_13 is a full-length clone, including the entire coding sequence of a secreted protein (also referred to herein as "pm4_13 protein"}.
The nucleotide sequence of pm4_13 as presently determined is reported in SEQ
ID N0:17, and includes a poly(A) tail. What applicants presently believe to be the proper 2 0 reading frame and the predicted amino acid sequence of the pm4_13 protein corresponding to the foregoing nucleotide sequence is reported in SEQ ID
N0:18. Amino acids 49 to 61 of SEQ ID N0:18 are a predicted leader/signal sequence, with the predicted mature amino acid sequence beginning at amino acid 62. Due to the hydrophobic nature of the predicted leader/signal sequence, it is likely to act as a transmembrane domain 2 5 should the predicted leader/signal sequence not be separated from the remainder of the pm4_13 protein.
The EcoRI/NotI restriction fragment obtainable from the deposit containing clone pm4_13 should be approximately 3100 bp.
The nucleotide sequence disclosed herein for pm4_13 was searched against the 3 0 GenBank and GeneSeq nucleotide sequence databases using BLASTN/BLASTX and FASTA search protocols. pm4_13 demonstrated at least some similarity with sequences identified as H98853 (yx15d10.s1 Homo sapiens cDNA clone 261811 3'), 843710 (yg19f12.s1 Homo sapiens cDNA clone 32692 3'), and T22962 (Human gene signature HUMGS04687). The predicted amino acid sequence disclosed herein for pm4_13 was searched against the GenPept and GeneSeq amino acid sequence databases using the BLASTX search protocol. The predicted pm4_13 protein demonstrated at least some similarity to sequences identified as U20865 (y1r246wp [Saccharomyces cerevisiae]), 229121 (ZK757.1 [Caenorhabditis elegansJ), 246259 (N0325 gene product [Saccharomyces cerevisiaeJ), 268507 (m18.8 [Caenorhabditis elegansJ), and 271602 (orf yn1326c [Saccharo-myces cerevisiaeJ). Based upon sequence similarity, pm4_13 proteins and each similar protein or peptide may share at least some activity. The TopPredII computer program predicts three additional potential transmembrane domains within the pm4_13 protein sequence, centered around amino acids 86,184, and 225 of SEQ ID N0:18, respectively.
Clone "pt326 4"
A polynucleotide of the present invention has been identified as clone "pt326 4".
pt326_4 was isolated from a human adult blood (lymphoblastic leukemia MOLT-4}
cDNA
library using methods which are selective for cDNAs encoding secreted proteins (see U.S.
Pat. No. 5,536,637), or was identified as encoding a secreted or transmembrane protein on the basis of computer analysis of the amino acid sequence of the encoded protein. pt326 4 is a full-length clone, including the entire coding sequence of a secreted protein (also referred to herein as "pt326_4 protein").
The nucleotide sequence of pt326 4 as presently determined is reported in SEQ
ID
2 0 N0:19, and includes a poly(A) tail. What applicants presently believe to be the proper reading frame and the predicted amino acid sequence of the pt326 4 protein corresponding to the foregoing nucleotide sequence is reported in SEQ ID
N0:20.
The EcoRI/NotI restriction fragment obtainable from the deposit containing clone pt326 4 should be approximately 3100 bp.
2 5 The nucleotide sequence disclosed herein for pt326_4 was searched against the GenBank and GeneSeq nucleotide sequence databases using BLASTN/BLASTX and FASTA search protocols. pt326_4 demonstrated at least some similarity with sequences identified as AA361324 (EST70660 T-cell lymphoma Homo sapiens cDNA 5' end), (yv19c12.s1 Homo sapiens cDNA clone 243190 3'), T22309 (Human gene signature 3 0 HUMGS03882), and U17907 (Human chromosome 17q21 mRNA clone B169). The predicted amino acid sequence disclosed herein for pt326 4 was searched against the GenPept and GeneSeq amino acid sequence databases using the BLASTX search protocol.
The predicted pt326 4 protein demonstrated at least some similarity to sequences identified as D89646 (GTBP-ALT [Homo sapiens]) and U28946 (G/T mismatch binding protein (GTBP) [Homo sapiensj). Based upon sequence similarity, pt326 4 proteins and each similar protein or peptide may share at least some activity.
posit of Clones Clones cc359 4, ct547 2, en553_l, nn296_2, nq27 13, pk65 4, pk855_1, PL776_6, pm4_13, and pt326 4 were deposited on March 31,1998 with the American Type Culture Collection (10801 University Boulevard, Manassas, Virginia 20110-2209 U.S.A.) as an original deposit under the Budapest Treaty and were given the accession number ATCC
98715, from which each clone comprising a particular polynucleotide is obtainable. All restrictions on the availability to the public of the deposited material will be irrevocably removed upon the granting of the patent, except for the requirements specified in 37 C.F.R. ~ 1.808(b), and the term of the deposit will comply with 37 C.F.R. ~
1.806.
Each clone has been transfected into separate bacterial cells (E. coli) in this composite deposit. Each clone can be removed from the vector in which it was deposited by performing an EcoRI/NotI digestion (5' site, EcoRI; 3' site, NotI) to produce the appropriate fragment for such clone. Each clone was deposited in either the pED6 or pNOTs vector depicted in Figures 1A and 1B, respectively. The pED6dpc2 vector ("pED6") was derived from pED6dpc1 by insertion of a new polylinker to facilitate cDNA cloning (Kaufman et al.,1991, Nucleic Acids Res. 19: 4485-4490); the pNOTs vector 2 0 was derived from pMT2 (Kaufman et al.,1989, Mol. Cell. Biol. 9: 946-958) by deletion of the DHFR sequences, insertion of a new polylinker, and insertion of the M13 origin of replication in the CIaI site. In same instances, the deposited clone can become "flipped"
(i.e., in the reverse orientation) in the deposited isolate. In such instances, the cDNA insert can still be isolated by digestion with EcoRI and NotI. However, NotI will then produce 2 5 the 5' site and EcoRI will produce the 3' site for placement of the cDNA
in proper orientation for expression in a suitable vector. The cDNA may also be expressed from the vectors in which they were deposited.
Bacterial cells containing a particular clone can be obtained from the composite deposit as follows:
3 0 An oligonucleotide probe or probes should be designed to the sequence that is known for that particular clone. This sequence can be derived from the sequences provided herein, or from a combination of those sequences. The sequence of an oligonucleotide probe that was used to isolate or to sequence each full-length clone is identified below, and should be most reliable in isolating the clone of interest.

ne Probe Sequence cc359 4 SEQ ID N0:21 c~~ 2 SEQ ID N0:22 en553_1 SEQ ID N0:23 nn296 2 SEQ ID N0:24 nq27 13 SEQ ID N0:25 Pk~-4 SEQ ID N0:26 pk855_1 SEQ ID N0:27 PL776_6 SEQ ID N0:28 pm4_13 SEQ ID N0:29 pt326_4 SEQ ID N0:30 In the sequences listed above which include an N at position 2, that position is occupied in preferred probes/primers by a biotinylated phosphoaramidite residue rather than a nucleotide (such as, for example, that produced by use of biotin phosphoramidite (1-dimethoxytrityloxy-2-(N-biotinyl-4-aminobutyl)-propyl-3-O-(2-cyanoethyl)-(N,N-diisopropyl)-phosphoramadite) (Glen Research, cat. no. 10-1953)).
The design of the oligonucleotide probe should preferably follow these parameters:
2 0 (a) It should be designed to an area of the sequence which has the fewest ambiguous bases ("N's'"), if any;
(b) It should be designed to have a T~, of approx. 80 ° C (assuming 2° for each A or T and 4 degrees for each G or C).
The oligonucleotide should preferably be labeled with y 32P ATP (specific activity 6000 Ci/mmole) and T4 polynucleotide kinase using commonly employed techniques for labeling oligonucleotides. Other labeling techniques can also be used.
Unincorporated label should preferably be removed by gel filtration chromatography or other established methods. The amount of radioactivity incorporated into the probe should be quantitated by measurement in a scintillation counter. Preferably, specific activity of the resulting 3 0 probe should be approximately 4e+6 dpm/pmole.
The bacterial culture containing the pool of full-length clones should preferably be thawed and 100 ~.zl of the stock used to inoculate a sterile culture flask containing 25 ml of sterile L-broth containing ampicillin at 100 ug/ml. The culture should preferably be grown to saturation at 37°C, and the saturated culture should preferably be diluted in fresh L-broth. Aliquots of these dilutions should preferably be plated to determine the dilution and volume which will yield approximately 5000 distinct and well-separated colonies on solid bacteriological media containing L-broth containing ampicillin at 100 lxg/ml and agar at 1.5% in a 150 mm petri dish when grown overnight at 37°C. Other known methods of obtaining distinct, well-separated colonies can also be employed.
Standard colony hybridization procedures should then be used to transfer the colonies to nitrocellulose filters and lyse, denature and bake them.
The filter is then preferably incubated at 65°C for 1 hour with gentle agitation in 6X SSC (20X stock is 175.3 g NaCI/liter, 88.2 g Na citrate/liter, adjusted to pH 7.0 with NaOH) containing 0.5% SDS,100 ug/ml of yeast RNA, and 10 mM EDTA
(approximately

10 mL per 150 mm filter). Preferably, the probe is then added to the hybridization mix at a concentration greater than or equal to 1e+6 dpm/mL. The filter is then preferably incubated at 65°C with gentle agitation overnight. The filter is then preferably washed in 500 mL of 2X SSC/0.5% SDS at room temperature without agitation, preferably followed by 500 mL of 2X SSC/0.1% SDS at room temperature with gentle shaking for 15 minutes.
A third wash with O.1X SSC/0.5% SDS at 65°C for 30 minutes to 1 hour is optional. The filter is then preferably dried and subjected to autoradiography for sufficient time to visualize the positives on the X-ray film. Other known hybridization methods can also be employed.
2 0 The positive colonies are picked, grown in culture, and plasmid DNA
isolated using standard procedures. The clones can then, be verified by restriction analysis, hybridization analysis, or DNA sequencing.
Fragments of the proteins of the present invention which are capable of exhibiting biological activity are also encompassed by the present invention. Fragments of the 2 5 protein may be in linear form or they may be cyclized using known methods, for example, as described in H.U. Saragovi, et al., Bio/Technology 1~( , 773-778 (1992) and in R.S.
McDowell, et al., J. Amer. Chem. Soc.1~ 9245-9253 {1992), both of which are incorporated herein by reference. Such fragments may be fused to carrier molecules such as immunoglobulins for many purposes, including increasing the valency of protein binding 3 0 sites. For example, fragments of the protein may be fused through "linker sequences to the Fc portion of an immunoglobulin. For a bivalent form of the protein, such a fusion could be to the Fc portion of an IgG molecule. Other immunoglobulin isotypes may also be used to generate such fusions. For example, a protein - IgM fusion would generate a decavalent form of the protein of the invention.

The present invention also provides both full-length and mature forms of the disclosed proteins. The full-length form of the such proteins is identified in the sequence listing by translation of the nucleotide sequence of each disclosed clone. The mature forms) of such protein may be obtained by expression of the disclosed full-length polynucleotide (preferably those deposited with ATCC) in a suitable mammalian cell or other host cell. The sequences) of the mature forms) of the protein may also be determinable from the amino acid sequence of the full-length form.
The present invention also provides genes corresponding to the polynucleotide sequences disclosed herein. "Corresponding genes" are the regions of the genome that are transcribed to produce the mRNAs from which cDNA polynucleotide sequences are derived and may include contiguous regions of the genome necessary for the regulated expression of such genes. Corresponding genes may therefore include but are not limited to coding sequences, 5' and 3' untranslated regions, alternatively spliced exons, introns, promoters, enhancers, and silencer or suppressor elements. The corresponding genes can be isolated in accordance with known methods using the sequence information disclosed herein. Such methods include the preparation of probes or primers from the disclosed sequence information for identification and/or amplification of genes in appropriate genomic libraries or other sources of genomic materials. An "isolated gene" is a gene that has been separated from the adjacent coding sequences, if any, present in the genome of 2 0 the organism from which the gene was isolated.
The chromosomal location corresponding to the polynucleotide sequences disclosed herein may also be determined, for example by hybridizing appropriately labeled polynucleotides of the present invention to chromosomes in situ. It may also be possible to determine the corresponding chromosomal location for a disclosed 2 5 polynucleotide by identifying significantly similar nucleotide sequences in public databases, such as expressed sequence tags (ESTs), that have already been mapped to particular chromosomal locations. For at least some of the polynucleotide sequences disclosed herein, public database sequences having at least some similarity to the polynucleotide of the present invention have been listed by database accession number.
3 0 Searches using the GenBank accession numbers of these public database sequences can then be performed at an Internet site provided by the National Center for Biotechnology Information having the address http://www.ncbi.nlm.nih.gov/UniGene/, in order to identify "UniGene clusters" of overlapping sequences. Many of the "UruGene clusters"
so identified will already have been mapped to particular chromosomal sites.

WO 99!50405 PCTNS99/06946 Organisms that have enhanced, reduced, or modified expression of the genes) corresponding to the polynucleotide sequences disclosed herein are provided.
The desired change in gene expression can be achieved through the use of antisense polynucleotides or ribozymes that bind and/or cleave the mRNA transcribed from the gene (Albert and Morris,1994, Trends Pluirmacol. Sci.15(7): 250-254; Lavarosky et al.,1997, Biochem. Mol. Med. 62(1):11-22; and liampel,1998, Prog. Nucleic Acid Res. Mol.
Biol. 58:1-39; all of which are incorporated by reference herein). Transgenic animals that have multiple copies of the genes) corresponding to the polynucleotide sequences disclosed herein, preferably produced by transformation of cells with genetic constructs that are stably maintained within the transformed cells and their progeny, are provided.
Transgenic animals that have modified genetic control regions that increase or reduce gene expression levels, or that change temporal or spatial patterns of gene expression, are also provided (see European Patent No. 0 649 464 Bl, incorporated by reference herein).
In addition, organisms are provided in which the genes) corresponding to the polynucleotide sequences disclosed herein have been partially or completely inactivated, through insertion of extraneous sequences into the corresponding genes) or through deletion of all or part of the corresponding gene(s). Partial or complete gene inactivation can be accomplished through insertion, preferably followed by imprecise excision, of transposable elements (Plasterk,1992, Bioessays 14(9): 629-633; Zwaal et al.,1993, Proc. Natl.
2 0 Acad. Sci. USA 90(16): 7431-7435; Clark et al.,1994, Proc. Natl. Acad.
Sci. USA 91(2): 719-722;
all of which are incorporated by reference herein), or through homologous recombination, preferably detected by positive/negative genetic selection strategies (Mansour et al.,1988, Nature 336: 348-352; U.S. Patent Nos. 5,464,764; 5,487,992; 5,627,059;
5,631,153; 5,614, 396;
5,616,491; and 5,679,523; all of which are incorporated by reference herein).
These 2 5 organisms with altered gene expression are preferably eukaryotes and more preferably are mammals. Such organisms are useful for. the development of non-human models for the study of disorders involving the corresponding gene(s), and for the development of assay systems for the identification of molecules that interact with the protein products) of the corresponding gene(s).
3 0 Where the protein of the present invention is membrane-bound (e.g., is a receptor), the present invention also provides for soluble forms of such protein. In such forms, part or all of the intracellular and transmembrane domains of the protein are deleted such that the protein is fully secreted from the cell in which it is expressed. The intracellular and transmembrane domains of proteins of the invention can be identified in accordance with known techniques for determination of such domains from sequence information.
For example, the TopPredII computer program can be used to predict the location of transmembrane domains in an amino acid sequence, domains which are described by the location of the center of the transmsmbrane domain, with at least ten transmembrane amino acids on each side of the reported central residue(s).
Proteins and protein fragments of the present invention include proteins with amino acid sequence lengths that are at least 25%(more preferably at least 50%, and most preferably at least 75%) of the length of a disclosed protein and have at least 60% sequence identity (more preferably, at least 75% identity; most preferably at least 90%
or 95%
identity) with that disclosed protein, where sequence identity is determined by comparing the amino acid sequences of the proteins when aligned so as to maximize overlap and identity while minimizing sequence gaps. Also included in the present invention are proteins and protein fragments that contain a segment preferably comprising 8 or more (more preferably 20 or more, most preferably 30 or more) contiguous amino atids that shares at least 75% sequence identity (more preferably, at least 85% identity;
most preferably at least 95% identity) with any such segment of any of the disclosed proteins.
In particular, sequence identity may be determined using WU-BLAST
(Washington University BLAST) version 2.0 software, which builds upon WU-BLAST
version 1.4, which in turn is based on the public domain NCBI-BLAST version 1.4 2 0 (Altschul and Gish,1996, Local alignment statistics, Doolittle ed., Methods in Enzymology 266: 460-480; Altschul et al., 1990, Basic local alignment search tool, Journal of Molecular Biology 215: 403-410; Gish and States, 1993, Identification of protein coding regions by database similarity search, Nature Genetics 3: 266-272; Karlin and Altschul, 1993, Applications and statistics for multiple high-scoring segments in molecular 2 5 sequences, Proc. Natl. Acad. Sci. USA 90: 5873-5877; all of which are incorporated by reference herein). WU-BLAST version 2.0 executable programs for several UNIX
platforms can be downloaded from ftp://blast.wustl.edu/blast/executables. The complete suite of search programs (BLASTP, BLASTN, BLASTX, TBLASTN, and TBLASTX) is provided at that site, in addition to several support programs. WU-BLAST 2.0 is 3 0 copyrighted and may not be sold or redistributed in any form or manner without the express written consent of the author; but the posted executables may otherwise be freely used for commercial, nonprofit, or academic purposes. In all search programs in the suite -- BLASTP, BLASTN, BLASTX, TBLASTN and TBLASTX -- the gapped alignment routines are integral to the database search itself, and thus yield much better sensitivity and selectivity while producing the more easily interpreted output. Gapping can optionally be fumed off in all of these programs, if desired. The default penalty (Q) for a gap of length one is Q=9 for proteins and BLASTP, and Q=10 for BLASTN, but may be changed to any integer value including zero, one through eight, nine, ten, eleven, twelve through twenty, twenty-one through fifty, fifty-one through one hundred, etc. The default per-residue penalty for extending a gap (R) is R=2 for proteins and BLASTP, and R=10 for BLASTN, but may be changed to any integer value including zero, one, two, three, four, five, six, seven, eight, nine, ten, eleven, twelve through twenty, twenty-one through fifty, fifty-one through one hundred, etc. Any combination of values for Q and R can be used in order to align sequences so as to maximize overlap and identity while minimizing sequence gaps.
The default amino acid comparison matrix is BLOSLTM62, but other amino acid comparison matrices such as PAM can be utilized.
Species homologues of the disclosed polynucleotides and proteins are also provided by the present invention. As used herein, a "species homologue" is a protein or polynucleotide with a different species of origin from that of a given protein or polynucleotide, but with significant sequence similarity to the given protein or polynucleotide. Preferably, polynucleotide species homologues have at least 60% sequence 2 0 identity (more preferably, at least 75% identity; most preferably at least 90% identity) with the given polynucleotide, and protein species homologues have at least 30%
sequence identity (more preferably, at least 45% identity; most preferably at least 60%
identity) with the given protein, where sequence identity is determined by comparing the nucleotide sequences of the polynucleotides or the amino acid sequences of the proteins when 2 5 aligned so as to maximize overlap and identity while minimizing sequence gaps. Species homologues may be isolated and identified by making suitable probes or primers from the sequences provided herein arid screening a suitable nucleic acid source from the desired species. Preferably, species homologues are those isolated from mammalian species. Most preferably, species homologues are those isolated from certain mammalian 3 0 species such as, for example, Pan troglodytes, Gorilla gorilla, Pongo pygmaeus, Hylobates concolor, Macaca mulatta, Papio papio, Papio humadryas, Cercopithecus aethiops, Cebus capucinus, Aotus trivirgatus, Sanguinus Oedipus, Microcebus murinus, Mus musculus, Rattus norvegicus, Cricetulus griseus, Felis catus, Mustela vison, Canis famiIiaris, Oryctolagus cuniculus, Bos taurus, Ovis aries, Sus scrofa, and Eguus caballus, for which genetic maps have been created allowing the identification of syntenic relationships between the genomic organization of genes in one species and the genomic organization of the related genes in another species (O'Brien and Seuanez, 1988, Ann. Rev. Genet. 22: 323-351; O'Brien et al., 1993, Nature Genetics 3:103-112; Johansson et al.,1995, Genomics 25: 682-690; Lyons et al.,1997, Nature Genetics 15: 47-56; O'Brien et al.,1997, Trends in Genetics 13(10): 393-399;
Carver and Stubbs, 1997, Genome Research 7:1123-1137; all of which are incorporated by reference herein).
The invention also encompasses allelic variants of the disclosed polynucleotides or proteins; that is, naturally-occurring alternative forms of the isolated polynucleotides which also encode proteins which are identical or have significantly similar sequences to those encoded by the disclosed polynudeotides. Preferably, allelic variants have at least 60% sequence identity (more preferably, at least 75% identity; most preferably at least 90%
identity) with the given polynucleotide, where sequence identity is determined by comparing the nucleotide sequences of the polynucleotides when aligned so as to maximize overlap and identity while minimizing sequence gaps. Allelic variants may be isolated and identified by making suitable probes or primers from the sequences provided herein and screening a suitable nucleic acid source from individuals of the appropriate species.
The invention also includes polynucleotides with sequences complementary to 2 0 those of the polynucleotides disclosed herein.
The present invention also includes polynucleotides that hybridize under reduced stringency conditions, more preferably stringent conditions, and most preferably highly stringent conditions, to polynucleotides described herein. Examples of stringency conditions are shown in the table below: highly stringent conditions are those that are at 2 5 least as stringent as, for example, conditions A-F; stringent conditions are at least as stringent as, for example, conditions G-L; and reduced stringency conditions are at least as stringent as, for example, conditions M-R.
Sl StringencyPolynucleotideHybridHybridization TemperatureWash ConditionHybrid Lengthand Temperature (bp)$ Buffers and Buffer' A DNA:DNA s SO 65C; lxSSC -or- 65C; 0.3xSSC
42C; lxSSC, 50% formamide B DNA:DNA <50 TB*; lxSSC TB*; ixSSC

C DNA:RNA 2 50 67C; lxSSC -or- 67C; 0.3xSSC
45C; lxSSC, 50% fonsnamide D DNA:RNA <50 Tp*; lxSSC Tp*; lxSSC

E RNA:RNA 2 SO 70C; lxSSC-or- 70C; 0.3xSSC
50C; lxSSC, 50% formamide F RNA:RNA <50 TF*; lxSSC TF*; lxSSC

G DNA:DNA 2 50 65C; 4xSSC -or- 65C; lxSSC
42C; 4xSSC, 50% formamide H DNA:DNA <50 TH*; 4xSSC TH*; 4xSSC

I DNA:RNA Z 50 67C; 4xSSC -or- 67C; lxSSC
45C; 4xSSC, 50% formamide J DNA:RNA <50 T~*; 4xSSC TJ*; 4xSSC

K RNA:RNA i 50 70C; 4xSSC -or- 67C; lxSSC
50C; 4xSSC, 50% formamide L RNA:RNA <50 T~*; 2xSSC T~*; 2xSSC

M DNA:DNA 2 50 50C; 4xSSC -or- 50C; 2xSSC
40C; 6xSSC, 50% fonmamide N DNA:DNA t50 TN*; 6xSSC TN*; 6xSSC

O DNA:RNA Z 50 55C; 4xSSC -or- 55C; 2xSSC
42C; 6xSSC, 50% formamide P DNA:RNA <50 Tp*; 6xSSC TP*; 6xSSC

Q RNA:RNA 2 50 60C; 4xSSC -or- 60C; 2xSSC
45C; 6xSSC, 50% formamide 2 R RNA:RNA <50 TR"; 4xSSC TR*; 4xSSC

t: The hybrid length is that antidpated for the hybridized regions) of the hybridizing polynucleotides. When hybridizing a polynucleotide to a target polynucleotide of unknown sequence, the hybrid length is assumed to be that of the hybridizing polynucleotide. When polynucleotides of known sequence are hybridized, the 2 5 hybrid length can be determined by aligning the sequences of the polynucleotides and identifying the region or regions of optimal sequence complementarity.
': SSPE (lxSSPE is 0.15M NaCI, lOmM NaHZPO" and 1.25mM EDTA, pH 7.4) can be substituted for SSC
(lxSSC is 0.15M NaCI and lSmM sodium citrate) in the hybridization and wash buffers; washes are performed for 15 minutes after hybridization is complete.
3 0 "TB - TR: The hybridization temperature for hybrids anticipated to be less than 50 base pairs in length should be 5-10°C less than the melting temperature (Tm) of the hybrid, where Tm is determined according to the following equations. For hybrids less than 18 base pairs in length, Tm(°C) = 2(# of A + T bases) + 4(# of G +
C bases). For hybrids between 18 and 49 base pairs in length, Tm(°C) =
81.5 + 16.6(log,o[Na']) + 0.41(%G+C) (600/N), where N is the number of bases in the hybrid, and [Na'] is the concentration of sodium ions in the 3 5 hybridization buffer ([Na'] for lxSSC = 0.165 M).

Additional examples of stringency conditions for polynucleotide hybridization are provided in Sambrook, J., E.F. Fritsch, and T. Maniatis, 1989, Molecular Cloning: A
Laboratory Manual, Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY, chapters 9 and 11, and Current Protocols in Molecular Biology,1995, F.M.
Ausubel et al., eds., John Wiley & Sons, Inc., sections 2.10 and 6.3-6.4, incorporated herein by reference.
Preferably, each such hybridizing polynucleotide has a length that is at least 25%(more preferably at least 50%, and most preferably at least 75%) of the length of the polynucleotide of the present invention to which it hybridizes, and has at least 60%
sequence identity (more preferably, at least 75% identity; most preferably at least 90% or 95% identity) with the polynucleotide of the present invention to which it hybridizes, where sequence identity is determined by comparing the sequences of the hybridizing polynucleotides when aligned so as to maximize overlap and identity while minimizing sequence gaps.
The isolated polynucleotide of the invention may be operably linked to an expression control sequence such as the pMT2 or pED expression vectors disclosed in Kaufman et al., Nucleic Acids Res. ~, 4485-4490 (1991), in order to produce the protein recombinantly. Many suitable expression control sequences are known in the art. General methods of expressing recombinant proteins are also known and are exemplified in R.
Kaufman, Methods in Enzymology 1~ 537-566 (1990). As defined herein "operably 2 0 linked" means that the isolated polynucleotide of the invention and an expression control sequence are situated within a vector or cell in such a way that the protein is expressed by a host cell which has been transformed (transfected) with the ligated polynucleotide/expression control sequence.
A number of types of cells may act as suitable host cells for expression of the 2 5 protein. Mammalian host cells include, for example, monkey COS cells, Chinese Hamster Ovary (CHO) cells, human kidney 293 cells, human epidermal A431 cells, human Co1o205 cells, 3T3 cells, CV-1 cells, other transformed primate cell lines, normal diploid cells, cell strains derived from in vi o culture of primary tissue, primary explants, HeLa cells, mouse L cells, BHK, HL-60, U937, HaK or jurkat cells.
3 0 Alternatively, it may be possible to produce the protein in lower eukaryotes such as yeast or in prokaryotes such as bacteria. Potentially suitable yeast strains include Saccharomyces cerevisiae, Schizosaccharomyces pombe, Kluyveromyces strains, Candida, or any yeast strain capable of expressing heterologous proteins. Potentially suitable bacterial strains include Escherichia coli, Bacillus subtilis, Salmonella typhimurium, or any bacterial strain capable of expressing heterologous proteins. If the protein is made in yeast or bacteria, it may be necessary to modify the protein produced therein, for example by phosphorylation or glycosylation of the appropriate sites, in order to obtain the functional protein. Such covalent attachments may be accomplished using known chemical or enzymatic methods.
The protein may also be produced by operably linking the isolated polynucleotide of the invention to suitable control sequences in one or more insect expression vectors, and employing an insect expression system. Materials and methods for baculovirus/insect cell expression systems are commercially available in kit form from, e.g., Invitrogen, San Diego, California, U.S.A. (the MaxBac~ kit), and such methods are well known in the art, as described in Summers and Smith, Texas A~;ricultural Experiment Station Bulletin No 1555 r198~, incorporated herein by reference. As used herein, an insect cell capable of expressing a polynucleotide of the present invention is "transformed."
The protein of the invention may be prepared by culturing transformed host cells under culture conditions suitable to express the recombinant protein. The resulting expressed protein may then be purified from such culture (i.e., from culture medium or cell extracts} using known purification processes, such as gel filtration and ion exchange chromatography. The purification of the protein may also include an affinity column 2 0 containing agents which will bind to the protein; one or more column steps over such affinity resins as concanavalin A-agarose, heparin-toyopearl~ or Cibacrom blue Sepharose0; one or more steps involving hydrophobic interaction chromatography using such resins as phenyl ether, butyl ether, or propyl ether; or immunoaffinity chromatography.
2 5 Alternatively, the protein of the invention may also be expressed in a form which will facilitate purification. For example, it may be expressed as a fusion protein, such as those of maltose binding protein (MBP), glutathione-S-transferase (GST) or thioredoxin (TRX). Kits for expression and purification of such fusion proteins are commercially available from New England BioLabs (Beverly, MA), Pharmacia (Piscataway, NJ) and 3 0 Invitrogen Corporation (Carlsbad, CA), respectively. The protein can also be tagged with an epitope and subsequently purified by using a specific antibody directed to such epitope. One such epitope ("Flag") is commercially available from the Eastman Kodak Company (New Haven, CT).

Finally, one or more reverse-phase high performance liquid chromatography (RP-I-iPLC) steps employing hydrophobic RP-HPLC media, e.g., silica gel having pendant methyl or other aliphatic groups, can be employed to further purify the protein. Some or all of the foregoing purification steps, in various combinations, can also be employed to provide a substantially homogeneous isolated recombinant protein. The protein thus purified is substantially free of other mammalian proteins and is defined in accordance with the present invention as an "isolated protein."
The protein of the invention may also be expressed as a product of transgenic animals, e.g., as a component of the milk of transgenic cows, goats, pigs, or sheep which are characterized by somatic or germ cells containing a nucleotide sequence encoding the protein.
The protein may also be produced by known conventional chemical synthesis.
Methods for constructing the proteins of the present invention by synthetic means are known to those skilled in the art. The synthetically-constructed protein sequences, by virtue of sharing primary, secondary or tertiary structural and/or conformational characteristics with proteins may possess biological properties in common therewith, including protein activity. Thus, they may be employed as biologically active or immunological substitutes for natural, purified proteins in screening of therapeutic compounds and in immunological processes for the development of antibodies.
2 0 The proteins provided herein also include proteins characterized by amino acid sequences similar to those of purified proteins but into which modification are naturally provided or deliberately engineered. For example, modifications in the peptide or DNA
sequences can be made by those skilled in the art using known techniques.
Modifications of interest in the protein sequences may include the alteration, substitution, replacement, 2 5 insertion or deletion of a selected amino acid residue in the coding sequence. For example, one or more of the cysteine residues may be deleted or replaced with another amino acid to alter the conformation of the molecule. Techniques for such alteration, substitution, replacement, insertion or deletion are well known to those skilled in the art (see, e.g., U.S. Patent No. 4,518,584). Preferably, such alteration, substitution, replacement, 3 0 insertion or deletion retains the desired activity of the protein.
Other fragments and derivatives of the sequences of proteins which would be expected to retain protein activity in whole or in part and may thus be useful for screening or other immunological methodologies may also be easily made by those skilled in the art given the disclosures herein. Such modifications are believed to be encompassed by the present invention.
USES AND BIOLOGICAL ACTIVITY
The polynucleotides and proteins of the present invention are expected to exhibit one or more of the uses or biological activities (including those associated with assays cited herein) identified below. Uses or activities described for proteins of the present invention may be provided by administration or use of such proteins or by administration or use of polynucleotides encoding such proteins (such as, for example, in gene therapies or vectors suitable for introduction of DNA).
Research s and Utilities The polynucleotides provided by the present invention can be used by the research community for various purposes. The polynucleotides can be used to express recombinant protein for analysis, characterization or therapeutic use; as markers for tissues in which the corresponding protein is preferentially expressed (either constitutively or at a particular stage of tissue differentiation or development or in disease states); as molecular weight markers on Southern gels; as chromosome markers or tags (when labeled) to identify chromosomes or to map related gene positions; to compare 2 0 with endogenous DNA sequences in patients to identify potential genetic disorders; as probes to hybridize and thus discover novel, related DNA sequences; as a source of information to derive PCR primers for genetic fingerprinting; as a probe to "subtract-out"
known sequences in the process of discovering other novel polynucleotides; for selecting and making oligomers for attachment to a "gene chip" or other support, including for 2 5 examination of expression patterns; to raise anti-protein antibodies using DNA
immunization techniques; and as an antigen to raise anti-DNA antibodies or elicit another immune response. Where the polynucleotide encodes a protein which binds or potentially binds to another protein (such as, for example, in a receptor-ligand interaction), the polynucleotide can also be used in interaction trap assays (such as, for example, those 3 0 described in Gyuris et al.,1993, Cell 75: 791-803 and in Rossi et al.,1997, Proc. Natl. Acad.
Sci. USA 94: 8405-8410, all of which are incorporated by reference herein) to identify polynucleotides encoding the other protein with which binding occurs or to identify inhibitors of the binding interaction.

The proteins provided by the present invention can similarly be used in assay to determine biological activity, including in a panel of multiple proteins for high-throughput screening; to raise antibodies or to elicit another immune response; as a reagent (including the labeled reagent) in assays designed to quantitatively determine levels of the protein (or its receptor) in biological fluids; as markers for tissues in which the corresponding protein is preferentially expressed (either constitutively or at a particular stage of tissue differentiation or development or in a disease state); and, of course, to isolate correlative receptors or ligands. Where the protein binds or potentially binds to another protein (such as, for example, in a receptor-ligand interaction), the protein can be used to identify the other protein with which binding occurs or to identify inhibitors of the binding interaction. Proteins involved in these binding interactions can also be used to screen for peptide or small molecule inhibitors or agorusts of the binding interaction.
Any or all of these research utilities are capable of being developed into reagent grade or kit format for commercialization as research products.
Methods for performing the uses listed above are well known to those skilled in the art. References disclosing such methods include without limitation "Molecular Cloning: A Laboratory Manual", 2d ed., Cold Spring Harbor Laboratory Press, Sambrook, J., E.F. Fritsch and T. Maruatis eds., 1989, and "Methods in Enzymology: Guide to 2 0 Molecular Cloning Techniques", Academic Press, Berger, S.L. and A.R.
Kimmel eds.,1987.
Nutritional Uses Polynucleotides and proteins of the present invention can also be used as nutritional sources or supplements. Such uses include without limitation use as a protein 2 5 or amino acid supplement, use as a carbon source, use as a nitrogen source and use as a source of carbohydrate. In such cases the protein or polynucleotide of the invention can be added to the feed of a particular organism or can be administered as a separate solid or liquid preparation, such as in the form of powder, pills, solutions, suspensions or capsules. In the case of microorganisms, the protein or polynucleotide of the invention 3 0 can be added to the medium in or on which the microorganism is cultured.
Cytokine and Cell Proliferation/Differentiation Activity A protein of the present invention may exhibit cytokine, cell proliferation (either inducing or inhibiting) or cell differentiation (either inducing or inhibiting) activity or may WO 99/50405 PCTNS99i06946 induce production of other cytokines in certain cell populations. Many protein factors discovered to date, including all known cytokines, have exhibited activity in one or more factor-dependent cell proliferation assays, and hence the assays serve as a convenient confirmation of cytokine activity. The activity of a protein of the present invention is evidenced by any one of a number of routine factor dependent cell proliferation assays for cell lines including, without limitation, 32D, DA2, DA1G, T10, B9, B9/11, BaF3, MC9/G, M+ (preB M+), 2E8, RBS, DA1,123, T1165, HT2, CTLL2, TF-1, Mole and CMK.
The activity of a protein of the invention may, among other means, be measured by the following methods:
Assays for T-cell or thymocyte proliferation include without limitation those described in: Current Protocols in Immunology, Ed by J. E. Coligan, A.M.
Kruisbeek, D.H.
Margulies, E.M. Shevach, W Strober, Pub. Greene Publishing Associates and Wiley-Interscience (Chapter 3, In Vitro assays for Mouse Lymphocyte Function 3.1-3.19; Chapter 7, Immunologic studies in Humans); Takai et al., J. Immunol. 137:3494-3500, 1986;
Bertagnolli et al., J. Irnmunol.145:1706-1712, 1990; Bertagnolli et al., Cellular Immunology 133:327-341, 1991; Bertagnolli, et al., J. Immunol. 149:3778-3783,1992; Bowman et al., J.
Immunol.152: 1756-1761,1994.
Assays for cytokine production and/or proliferation of spleen cells, lymph node cells or thymocytes include, without limitation, those described in:
Polyclonal T cell 2 0 stimulation, Kruisbeek, A.M. and Shevach, E.M. In Current Protocols in Immunology. J.E.e.a.
Coligan eds. Vol 1 pp. 3.12.1-3.12.14, John Wiley and Sons, Toronto. 1994; and Measurement of mouse and human Interferon Y, Schreiber, R.D. In Current Protocols in Immunology. J.E.e.a. Coligan eds. Vol 1 pp. 6.8.1-6.8.8, John Wiley and Sons, Toronto.1994.
Assays for proliferation and differentiation of hernatopoietic and lymphopoietic 2 5 cells include, without limitation, those described in: Measurement of Human and Murine Interleukin 2 and Interleukin 4, Bottomiy, K., Davis, L.S. and Lipsky, P.E. In Current Protocols in Immunology. J.E.e.a. Coligan eds. Vol 1 pp. 6.3.1-6.3.12, John Wiley and Sons, Toronto. 1991; deVries et al., J. Exp. Med. 173:1205-1211, 1991; Moreau et al., Nature 336:690-692, 1988; Greenberger et al., Proc. Natl. Acad. Sci. U.S.A. 80:2931-2938, 1983;
3 0 Measurement of mouse and human interleukin 6 - Nordan, R. In Current Protocols in Immunology. J.E.e.a. Coligan eds. Vol 1 pp. 6.6.1-6.6.5, John Wiley and Sons, Toronto.1991;
Smith et al., Proc. Natl. Acad. Sci. U.S.A. 83:1857-1861, 1986; Measurement of human Interleukin 11- Benrtett, F., Giannotti, J., Clark, S.C. and Turner, K. J. In Current Protocols in Immunology. J.E.e.a. Coligan eds. Vol 1 pp. 6.15.1 John Wiley and Sons, Toronto. 1991;

Measurement of mouse and human lnterleukin 9 - Ciarletta, A., Giannotti, J., Clark, S.C.
and Turner, K.J. In Current Protocols in Immunology. J.E.e.a. Coligan eds. Vol 1 pp. 6.13.1, John Wiley and Sons, Toronto. 1991.
Assays for T-cell clone responses to antigens (which will identify, among others, proteins that affect APC-T cell interactions as well as direct T-cell effects by measuring proliferation and cytokine production) include, without limitation, those described in:
Current Protocols in Immunology, Ed by J. E. Coligan, A.M. Kruisbeek, D.H.
Margulies, E.M. Shevach, W Strober, Pub. Greene Publishing Associates and Wffey-Interscience (Chapter 3, In Vitro assays for Mouse Lymphocyte Function; Chapter 6, Cytokines and their cellular receptors; Chapter 7, Immunologic studies in Humans);
Weinberger et al., Proc. Natl. Acad. Sci. USA 77:6091-6095, 1980; Weinberger et al., Eur. J.
Immun.

11:405-411, 1981; Takai et al., J. Immunol. 137:3494-3500, 1986; Takai et al., J. Immunol.
140:508-512, 1988.
Immune Stimulating or Sup ressing Activity A protein of the present invention may also exhibit immune stimulating or immune suppressing activity, including without limitation the activities for which assays are described herein. A protein may be useful in the treatment of various immune deficiencies and disorders (including severe combined immunodeficiency (SCID)), e.g., 2 0 in regulating (up or down) growth and proliferation of T and/or B
lymphocytes, as well as effecting the cytolytic activity of NK cells and other cell populations.
These immune deficiencies may be genetic or be caused by viral (e.g., HIV) as well as bacterial or fungal infections, or may result from autoimmune disorders. More specifically, infectious diseases causes by viral, bacterial, fungal or other infection may be treatable using a 2 5 protein of the present invention, including infections by HIV, hepatitis viruses, herpesviruses, mycobacteria, Leishmania spp., malaria spp. and various fungal infections such as candidiasis. Of course, in this regard, a protein of the present invention may also be useful where a boost to the immune system generally may be desirable, i.e., in the treatment of cancer.
3 0 Autoimmune disorders which may be treated using a protein of the present invention include, for example, connective tissue disease, multiple sclerosis, systemic lupus erythematosus, rheumatoid arthritis, autoimmune pulmonary inflammation, Guillain-Barre syndrome, autoimmune thyroiditis, insulin dependent diabetes mellitis, myasthenia gravis, graft-versus-host disease and autoimmune inflammatory eye disease.

Such a protein of the present invention may also to be useful in the treatment of allergic reactions and conditions, such as asthma (particularly allergic asthma) or other respiratory problems. Other conditions, in which immune suppression is desired (including, for example, organ transplantation), may also be treatable using a protein of the present invention.
Using the proteins of the invention it may also be possible to regulate immune responses in a number of ways. Down regulation may be in the form of inhibiting or blocking an immune response already in progress or may involve preventing the induction of an immune response. The functions of activated T cells may be inhibited by suppressing T cell responses or by inducing specific tolerance in T cells, or both.
Immunosuppression of T cell responses is generally an active, non-antigen-specific, process which requires continuous exposure of the T cells to the suppressive agent.
Tolerance, which involves inducing non-responsiveness or anergy in T cells, is distinguishable from immunosuppression in that it is generally antigen-specific and persists after exposure to the tolerizing agent has ceased. Operationally, tolerance can be demonstrated by the lack of a T cell response upon reexposure to specific antigen in the absence of the tolerizing agent.
Down regulating or preventing one or more antigen functions (including without limitation B lymphocyte antigen functions (such as , for example, B7)), e.g., preventing 2 0 high level lymphokine synthesis by activated T cells, will be useful in situations of tissue, skin and organ transplantation and in graft-versus-host disease (GVHD). For example, blockage of T cell function should result in reduced tissue destruction in tissue transplantation. Typically, in tissue transplants, rejection of the transplant is initiated through its recognition as foreign by T cells, followed by an immune reaction that destroys 2 5 the transplant. The administration of a molecule which inhibits or blocks interaction of a B7 lymphocyte antigen with its natural ligand(s) on immune cells (such as a soluble, monomeric form of a peptide having B7-2 activity alone or in conjunction with a monomeric form of a peptide having an activity of another B lymphocyte antigen (e.g., B7-1, B7-3) or blocking antibody), prior to transplantation can lead to the binding of the 3 0 molecule to the natural ligand(s) on the immune cells without transmitting the corresponding costimulatory signal. Blocking B lymphocyte antigen function in this matter prevents cytokine synthesis by immune cells, such as T cells, and thus acts as an immunosuppressant. Moreover, the lack of costimulation may also be sufficient to anergize the T cells, thereby inducing tolerance in a subject. Induction of long-term tolerance by B lymphocyte antigen-blocking reagents may avoid the necessity of repeated administration of these blocking reagents. To achieve sufficient immunosuppression or tolerance in a subject, it may also be necessary to block the function of a combination of B lymphocyte antigens.
The efficacy of particular blocking reagents in preventing organ transplant rejection or GVHD can be assessed using animal models that are predictive of efficacy in humans. Examples of appropriate systems which can be used include allogeneic cardiac grafts in rats and xenogeneic pancreatic islet cell grafts in mice, both of which have been used to examine the immunosuppressive effects of CTLA4Ig fusion proteins in vivo as described in Lenschow et al., Science 257:789-792 (1992) and Turka et al., Proc. Natl. Acad.
Sci USA, 89:11102-11105 (1992). In addition, marine models of GVHD (see Paul ed., Fundamental Immunology, Raven Press, New York, 1989, pp. 846-847) can be used to determine the effect of blocking B lymphocyte antigen function in vivo on the development of that disease.
Blocking antigen function may also be therapeutically useful for treating autoimmune diseases. Many autoimmune disorders are the result of inappropriate activation of T cells that are reactive against self tissue and which promote the production of cytokines and autoantibodies involved in the pathology of the diseases.
Preventing the activation of autoreactive T cells may reduce or eliminate disease symptoms.
2 0 Administration of reagents which block costimulation of T cells by disrupting receptor:ligand interactions of B lymphocyte antigens can be used to inhibit T
cell activation and prevent production of autoantibodies or T cell-derived cytokines which may be involved in the disease process. Additionally, blocking reagents may induce antigen-specific tolerance of autoreactive T cells which could lead to long-term relief from the disease. The efficacy of blocking reagents in preventing or alleviating autoimmune disorders can be determined using a number of well-characterized animal models of human autoimmune diseases. Examples include marine experimental autoimmune encephalitis, systemic lupus erythmatosis in MRL/lpr/Ipr mice or NZB hybrid mice, marine autoimmune collagen arthritis, diabetes mellitus in NOD mice and BB
rats, and 3 0 marine experimental myasthenia gravis (see Paul ed., Fundamental Immunology, Raven Press, New York,1989, pp. 840-856).
Upregulation of an antigen function (preferably a B lymphocyte antigen function), as a means of up regulating immune responses, may also be useful in therapy.
Upregulation of immune responses may be in the form of enhancing an existing immune response or eliciting an initial immune response. For example, enhancing an immune response through stimulating B lymphocyte antigen function may be useful in cases of viral infection. In addition, systemic viral diseases such as influenza, the common cold, and encephalitis might be alleviated by the administration of stimulatory forms of B
lymphocyte antigens systemically.
Alternatively, anti-viral immune responses may be enhanced in an infected patient by removing T cells from the patient, costimulating the T cells in vitro with viral antigen-pulsed APCs either expressing a peptide of the present invention or together with a stimulatory form of a soluble peptide of the present invention and reintroducing the in vitro activated T cells into the patient. Another method of enhancing anti-viral immune responses would be to isolate infected cells from a patient, transfect them with a nucleic acid encoding a protein of the present invention as described herein such that the cells express all or a portion of the protein on their surface, and reintroduce the transfected cells into the patient. The infected cells would now be capable of delivering a costimulatory signal to, and thereby activate, T cells in vivo.
In another application, up regulation or enhancement of antigen function (preferably B lymphocyte antigen function) may be useful in the induction of tumor immunity. Tumor cells (e.g., sarcoma, melanoma, lymphoma, leukemia, neuroblastoma, carcinoma) transfected with a nucleic acid encoding at least one peptide of the present 2 0 invention can be administered to a subject to overcome tumor-specific tolerance in the subject. If desired, the tumor cell can be transfected to express a combination of peptides.
For example, tumor cells obtained from a patient can be transfected ex vivo with an expression vector directing the expression of a peptide having B7-2-like activity alone, or in conjunction with a peptide having B7-1-like activity and/or B7-3-like activity. The transfected tumor cells are returned to the patient to result in expression of the peptides on the surface of the transfected cell. Alternatively, gene therapy techniques can be used to target a tumor cell for transfection in vivo.
The presence of the peptide of the present invention having the activity of a B
lymphocyte antigens) on the surface of the tumor cell provides the necessary 3 0 costimulation signal to T cells to induce a T cell mediated immune response against the transfected tumor cells. In addition, tumor cells which lack MHC class I or MHC class II
molecules, or which fail to reexpress sufficient amounts of MHC class I or MHC
class II
molecules, can be transfected with nucleic acid encoding all or a portion of (e.g., a cytoplasmic-domain truncated portion) of an MHC class I a chain protein and ~3z microglobulin protein or an MHC class II a chain protein and an MHC class II
~i chain protein to thereby express MHC class I or MHC class II proteins on the cell surface.
Expression of the appropriate class I or class II MHC in conjunction with a peptide having the activity of a B lymphocyte antigen (e.g., B7-1, B7-2, B7 3) induces a T
cell mediated immune response against the transfected tumor cell. Optionally, a gene encoding an antisense construct which blocks expression of an MHC class II associated protein, such as the invariant chain, can also be cotransfected with a DNA encoding a peptide having the activity of a B lymphocyte antigen to promote presentation of tumor associated antigens and induce tumor specific immunity. Thus, the induction of a T cell mediated immune response in a human subject may be sufficient to overcome tumor-specific tolerance in the subject.
The activity of a protein of the invention may, among other means, be measured by the following methods:
Suitable assays for thymocyte or splenocyte cytotoxicity include, without limitation, those described in: Current Protocols in Immunology, Ed by J. E.
Coligan, A.M.
ICruisbeek, D.H. Margulies, E.M. Shevach, W Strober, Pub. Greene Publishing Associates and Wiley-Interscience (Chapter 3, In Vitro assays for Mouse Lymphocyte Function 3.1-3.19; Chapter 7, Immunologic studies in Humans); Herrmann et al., Proc. Natl.
Acad. Sci.
USA 78:2488-2492,1981; Herrmann et al., J. Immunol.128:1968-1974,1982; Handa et al., 2 0 J. Immunol.135:1564-1572,1985; Takai et al., J. )mmunol.137:3494-3500,1986; Takai et al., J. Immunol.140:508-512, 1988; Herrmann et al., Proc. Natl. Acad. Sci. USA
78:2488-2492, 1981; Hemnann et al., J. lmmunol. 128:1968-1974, 1982; Handa et al., J.
Immunol.
135:1564-1572, 1985; Takai et al., J. Immunol. 137:3494-3500, 1986; Bowmanet al., J.
Virology 61:1992-1998; Takai et al., J. Immunol. 140:508-512, 1988;
Bertagnolli et aL, 2 5 Cellular Immunology 133:327-341,1991; Brown et al., J. Lmmunol.153:3079-3092, 1994.
Assays for T-cell-dependent immunoglobulin responses and isotype switching (which will identify, among others, proteins that modulate T-cell dependent antibody responses and that affect Th1 /Th2 profiles) include, without limitation, those described in: Maliszewski, J. Immunol.144:3028-3033,1990; and Assays for B cell function: In vitro 3 0 antibody production, Mond, J.J. and Brunswick, M. In Current Protocols in Immunology.
J.E.e.a. Coligan eds. Vol 1 pp. 3.8.1-3.8.16, John Wiley and Sons, Toronto.1994.
Mixed lymphocyte reaction (MLR) assays (which will identify, among others, proteins that generate predominantly Thl and CTL responses) include, without limitation, those described in: Current Protocols in Immunology, Ed by J. E. Coligan, A.M.
ICruisbeek, D.H. Margulies, E.M. Shevach, W Strober, Pub. Greene Publishing Associates and Wiley-Interscience (Chapter 3, In Vitro assays for Mouse Lymphocyte Function 3.1-3.19; Chapter 7, Immunologic studies in Humans); Takai et al., J. Immunol. 137:3494-3500,1986; Takai et al., J. Immunol.140:508-512, 1988; Bertagnolli et al., J. Immunol.149:3778-3783,1992.
Dendritic cell-dependent assays (which will identify, among others, proteins expressed by dendritic cells that activate naive T-cells) include, without limitation, those described in: Guery et al., J. Immunol. 134:536-544, 1995; Inaba et al., Journal of Experimental Medicine 173:549-559, 1991; Macatonia et al., Journal of Immunology 154:5071-5079,1995; Porgador et al., Journal of Experimental Medicine 182:255-260,1995;
Nair et al., Journal of Virology 67:4062-4069, 1993; Huang et al., Science 264:961-965, 1994; Macatonia et al., Journal of Experimental Medicine 169:1255-1264,1989;
Bhardwaj et aL, Journal of Clinical Investigation 94:797-807, 1994; and Inaba et al., Journal of Experimental Medicine 172:631-640,1990.
Assays for lymphocyte survival/apoptosis (which will identify, among others, proteins that prevent apoptosis after superantigen induction and proteins that regulate lymphocyte homeostasis) include, without limitation, those described in:
Darzynkiewicz et al., Cytometry 13:795-808,1992; Gorczyca et al., Leukemia 7:659-670,1993;
Gorczyca et al., Cancer Research 53:1945-1951, 1993; Itoh et al., Cell 66:233-243, 1991;
Zacharchuk, journal of Immunology 145:4037-4045, 1990; Zamai et al., Cytometry 14:891-897, 1993;
2 0 Gorczyca et al., International Journal of Oncology 1:639-648,1992.
Assays for proteins that influence early steps of T-cell commitment and development include, without limitation, those described in: Antica et al., Blood 84:111-117, 1994; Fine et al., Cellular Immunology 155:111-122,1994; Galy et al., Blood 85:2770-2778,1995; Toki et al., Proc. Nat. Acad Sci. USA 88:7548-7551,1991.
Hematgpoiesis Regulating Activ'~ty A protein of the present invention may be useful in regulation of hematopoiesis and, consequently, in the treatment of myeloid or lymphoid cell deficiencies.
Even marginal biological activity in support of colony forming cells or of factor-dependent cell 3 0 lines indicates involvement in regulating hematopoiesis, e.g. in supporting the growth and proliferation of erythroid progenitor cells alone or in combination with other cytokines, thereby indicating utility, for example, in treating various anemias or for use in conjunction with irradiation/chemotherapy to stimulate the production of erythroid precursors and/or erythroid cells; in supporting the growth and proliferation of myeloid cells such as granulocytes and monocytes/macrophages (i.e., traditional CSF
activity) useful, for example, in conjunction with chemotherapy to prevent or treat consequent myelo-suppression; in supporting the growth and proliferation of megakaryocytes and consequently of platelets thereby allowing prevention or treatment of various platelet disorders such as thrombocytopenia, and generally for use in place of or complimentary to platelet transfusions; and/or in supporting the growth and proliferation of hematopoietic stem cells which are capable of maturing to any and all of the above-mentioned hematopoietic cells and therefore find therapeutic utility in various stem cell disorders (such as those usually treated with transplantation, including, without limitation, aplastic anemia and paroxysmal nocturnal hemoglobinuria), as well as in repopulating the stem cell compartment post irradiation/chemotherapy, either in-vivo or ex-vivo (i.e., in conjunction with bone marrow transplantation or with peripheral progenitor cell transplantation (homologous or heterologous)) as normal cells or genetically manipulated for gene therapy.
The activity of a protein of the invention may, among other means, be measured by the following methods:
Suitable assays for proliferation and differentiation of various hematopoietic lines are cited above.
Assays for embryonic stem cell differentiation (which will identify, among others, 2 0 proteins that influence embryonic differentiation hematopoiesis) include, without limitation, those described in: Johansson et al. Cellular Biology 15:141-151,1995; Keller et al., Molecular and Cellular Biology 13:473-486, 1993; McClanahan et al., Blood 81:2903-2915,1993.
Assays for stem cell survival and differentiation (which will identify, among 2 5 others, proteins that regulate lympho-hematopoiesis) include, without limitation, those described in: Methylcellulose colony forming assays, Freshney, M.G. In Culture of Hematopoietic Cells. R.I. Freshney, et al. eds. Vol pp. 265-268, Wiley-Liss, Inc., New York, NY. 1994; Hirayama et al., Proc. Natl. Acad. Sci. USA 89:5907-5911, 1992;
Primitive hematopoietic colony forming cells with high proliferative potential, McNiece, LK. and 3 0 Briddell, R.A. In Culture of Hematopoietic Cells. R.I. Freshney, et al.
eds. Vol pp. 23-39, Wiley-Liss, Inc., New York, NY.1994; Neben et al., Experimental Hematology 22:353-359, 1994; Cobblestone area forming cell assay, Ploemacher, R.E. In Culture of Hematopoietic Cells. R.I. Freshney, et al. eds. Vol pp. 1-21, Wiley-Liss, Inc.., New York, NY.1994; Long term bone marrow cultures in the presence of stromal cells, Spooncer, E., Dexter, M. and Allen, T. In Culture of Hematopoietic Cells. R.I. Freshney, et al. eds. Vol pp. 163-179, Wiley-Liss, Inc., New York, NY.1994; Long term culture initiating cell assay, Sutherland, H.J. In Culture of Hematapoietic Cells. R.I. Freshney, et al. eds. Vol pp.139-162, Wiley-Liss, Inc., New York, NY.1994.
Tissue Growth ActivitX
A protein of the present invention also may have utility in compositions used for bone, cartilage, tendon, ligament and/or nerve tissue growth or regeneration, as well as for wound healing and tissue repair and replacement, and in the treatment of burns, incisions and ulcers.
A protein of the present invention, which induces cartilage and/or bone growth in circumstances where bone is not normally formed, has application in the healing of bone fractures and cartilage damage or defects in humans and other animals.
Such a preparation employing a protein of the invention may have prophylactic use in closed as well as open fracture reduction and also in the improved fixation of artificial joints. De novo bone formation induced by an osteogenic agent contributes to the repair of congenital, trauma induced, or oncologic resection induced craniofacial defects, and also is useful in cosmetic plastic surgery.
A protein of this invention may also be used in the treatment of periodontal 2 0 disease, and in other tooth repair processes. Such agents may provide an environment to attract bone-forming cells, stimulate growth of bone-forming cells or induce differentiation of progenitors of bone-forming cells. A protein of the invention may also be useful in the treatment of osteoporosis or osteoarthritis, such as through stimulation of bone and/or cartilage repair or by blocking inflammation or processes of tissue 2 5 destruction (collagenase activity, osteoclast activity, etc.) mediated by inflammatory processes.
Another category of tissue regeneration activity that may be attributable to the protein of the present invention is tendon/ligament formation. A protein of the present invention, which induces tendon/ligament-like tissue or other tissue formation in 3 0 circumstances where such tissue is not normally formed, has application in the healing of tendon or ligament tears, deformities and other tendon or ligament defects in humans and other animals. Such a preparation employing a tendon/ligament-like tissue inducing protein may have prophylactic use in preventing damage to tendon or ligament tissue, as well as use in the improved fixation of tendon or ligament to bone or other tissues, and in repairing defects to tendon or ligament tissue. De novo tendon/ligament-like tissue formation induced by a composition of the present invention contributes to the repair of congenital, trauma induced, or other tendon or ligament defects of other origin, and is also useful in cosmetic plastic surgery for attachment or repair of tendons or ligaments. The compositions of the present invention may provide an environment to attract tendon- or ligament-forming cells, stimulate growth of tendon- or ligament-forming cells, induce differentiation of progenitors of tendon- or ligament-forming cells, or induce growth of tendon/ligament cells or progenitors ex vivo for return in vivo to effect tissue repair. The compositions of the invention may also be useful in the treatment of tendinitis, carpal tunnel syndrome and other tendon or ligament defects. The compositions may also include an appropriate matrix and/or sequestering agent as a carrier as is well known in the art.
The protein of the present invention may also be useful for proliferation of neural cells and for regeneration of nerve and brain tissue, i.e. for the treatment of central and peripheral nervous system diseases and neuropathies, as well as mechanical and traumatic disorders, which involve degeneration, death or trauma to neural cells or nerve tissue. More specifically, a protein may be used in the treatment of diseases of the peripheral nervous system, such as peripheral nerve injuries, peripheral neuropathy and localized neuropathies, and central nervous system diseases, such as Alzheimer's, 2 0 Parkinson's disease, Huntingtori s disease, amyotrophic lateral sclerosis, and Shy-Drager syndrome. Further conditions which may be treated in accordance with the present invention include mechanical and traumatic disorders, such as spinal cord disorders, head trauma and cerebrovascular diseases such as stroke. Peripheral neuropathies resulting from chemotherapy or other medical therapies may also be treatable using a protein of the 2 5 invention.
Proteins of the invention may also be useful to promote better or faster closure of non-healing wounds, including without limitation pressure ulcers, ulcers associated with vascular insufficiency, surgical and traumatic wounds, and the like.
It is expected that a protein of the present invention may also exhibit activity for 3 0 generation or regeneration of other tissues, such as organs (including, for example, pancreas, liver, intestine, kidney, skin, endothelium), muscle (smooth, skeletal or cardiac) and vascular (including vascular endothelium) tissue, or for promoting the growth of cells comprising such tissues. Part of the desired effects may be by inhibition or modulation of fibrotic scarring to allow normal tissue to regenerate. A protein of the invention may also exhibit angiogenic activity.
A protein of the present invention may also be useful for gut protection or regeneration and treatment of lung or liver fibrosis, reperfusion injury in various tissues, and conditions resulting from systemic cytokine damage.
A protein of the present invention may also be useful for promoting or inhibiting differentiation of tissues described above from precursor tissues or cells; or for inhibiting the growth of tissues described above.
The activity of a protein of the invention may, among other means, be measured by the following methods:
Assays for tissue generation activity include, without limitation, those described in: International Patent Publication No. W095/16035 (bone, cartilage, tendon);
International Patent Publication No. W095/05846 (nerve, neuronal);
International Patent Publication No. W091 /07491 (skin, endothelium ).
Assays for wound healing activity include, without limitation, those described in:
Winter, epidermal Wound Healing,, pps. 71-112 (Maibach, HI and Rovee, DT, eds.), Year Book Medical Publishers, lnc., Chicago, as modified by Eagistein and Mertz, J.
Invest.
Dermatol 71:382-84 (1978).
2 0 Activin/Inhibin ActivitX
A protein of the present invention may also exhibit activin- or inhibin-related activities. Inhibins are characterized by their ability to inhibit the release of follicle stimulating hormone (FSH), while activins and are characterized by their ability to stimulate the release of follicle stimulating hormone (FSH). Thus, a protein of the present 2 5 invention, alone or in heterodimers with a member of the inhibin a family, may be useful as a contraceptive based on the ability of inhibins to decrease fertility in female mammals and decrease spermatogenesis in male mammals. Administration of sufficient amounts of other inhibins can induce infertility in these mammals. Alternatively, the protein of the invention, as a homodimer or as a heterodimer with other protein subunits of the inhibin-3 0 ~i group, may be useful as a fertility inducing therapeutic, based upon the ability of activin molecules in stimulating FSH release from cells of the anterior pituitary.
See, for example, United States Patent 4,798,885. A protein of the invention may also be useful for advancement of the onset of fertility in sexually immature mammals, so as to increase the lifetime reproductive performance of domestic animals such as cows, sheep and pigs.

The activity of a protein of the invention may, among other means, be measured by the following methods:
Assays for activin/inhibin activity include, without limitation, those described in:
Vale et al., Endocrinology 91:562-572,1972; Ling et al., Nature 321:779-782,1986; Vale et al., Nature 321:776-779,1986; Mason et al., Nature 318:659-663,1985; Forage et al., Proc.
Natl. Acad. Sci. USA 83:3091-3095,1986.
~emotactic /Chemokingtic Activity A protein of the present invention may have chemotactic or chemokineiic activity (e.g., act as a chemokine) fox mammalian cells, including, for example, monocytes, fibroblasts, neutrophils, T-cells, mast cells, eosinophils, epithelial and/or endothelial cells.
Chemotactic and chemokinetic proteins can be used to mobilize or attract a desired cell population to a desired site of action. Chemotactic or chemokinetic proteins provide particular advantages in treatment of wounds and other trauma to tissues, as well as in treatment of localized infections. For example, attraction of lymphocytes, monocytes or neutrophils to tumors or sites of infection may result in improved immune responses against the tumor or infecting agent.
A protein or peptide has chemotactic activity for a particular cell population if it can stimulate, directly or indirectly, the directed orientation or movement of such cell 2 0 population. Preferably, the protein or peptide has the ability to directly stimulate directed movement of cells. Whether a particular protein has chemotactic activity for a population of cells can be readily determined by employing such protein or peptide in any known assay for cell chemotaxis.
The activity of a protein of the invention may, among other means, be measured 2 5 by the following methods:
Assays for chemotactic activity (which will identify proteins that induce or prevent chemotaxis) consist of assays that measure the ability of a protein to induce the migration of cells across a membrane as well as the ability of a protein to induce the adhesion of one cell population to another cell population. Suitable assays for movement and adhesion 3 0 include, without limitation, those described in: Current Protocols in Immunology, Ed by J.E. Coligan, A.M. Kruisbeek, D.H. Margulies, E.M. Shevach, W.Strober, Pub.
Greene Publishing Associates and Wiley-Interscience (Chapter 6.12, Measurement of alpha and beta Chemokines 6.12.1-6.12.28; Taub et al. J. Clin. Invest. 95:1370-1376,1995; Lind et al.

APMIS 103:140-146,1995; Muller et al Eur. J. Immunol. 25: 1744-1748; Gruber et al. J. of Immunol.152:5860-5867,1994; Johnston et al. J. of Immunol. 153:1762-1768,1994.
Hemostatic and Thrombolytic Activit~r A protein of the invention may also exhibit hemostatic or thrombolytic activity.
As a result, such a protein is expected to be useful in treatment of various coagulation disorders (including hereditary disorders, such as hemophilias) or to enhance coagulation and other hemostatic events in treating wounds resulting from trauma, surgery or other causes. A protein of the invention may also be useful for dissolving or inhibiting formation of thromboses and for treatment and prevention of conditions resulting therefrom (such as, for example, infarction of cardiac and central nervous system vessels (e.g., stroke).
The activity of a protein of the invention may, among other means, be measured by the following methods:
Assay for hemostatic and thrombolytic activity include, without limitation, those described in: Linet et al., J. Clin. Pharmacol. 26:131-140,1986; Burdick et al., Thrombosis Res. 45:413-419,1987; Humphrey et al., Fibrinolysis 5:71-79 (1991); Schaub, Prostaglandins 35:467-474,1988.
~gptor/L~gand Activity A protein of the present invention may also demonstrate activity as receptors, receptor ligands or inhibitors or agonists of receptor/ligand interactions.
Examples of such receptors and ligands include, without limitation, cytokine receptors and their ligands, receptor kinases and their ligands, receptor phosphatases and their ligands, 2 5 receptors involved in cell-cell interactions and their ligands (including without limitation, cellular adhesion molecules (such as selectins, integrins and their ligands) and receptor/ligand pairs involved in antigen presentation, antigen recognition and development of cellular and humoral immune responses). Receptors and ligands are also useful for screening of potential peptide or small molecule inhibitors of the relevant 3 0 receptor/ligand interaction. A protein of the present invention (including, without limitation, fragments of receptors and ligands) may themselves be useful as inhibitors of receptor/ligand interactions.
The activity of a protein of the invention may, among other means, be measured by the following methods:

Suitable assays for receptor-ligand activity include without limitation those described in:Current Protocols in Immunology, Ed by J.E. Coligan, A.M.
Kruisbeek, D.H.
Margulies, E.M. Shevach, W.Strober, Pub. Greene Publishing Associates and Wiley-Interscience (Chapter 7.28, Measurement of Cellular Adhesion under static conditions 7.28.1-7.28.22), Takai et al., Proc. Natl. Acad. Sci. USA 84:6864-6868, 1987;
Bierer et al., J. Exp. Med.168:1145-1156, 1988; Rosenstein et al., J. Exp.
Med.169:149-160 1989; Stoltenborg et al., J. Immunol. Methods 175:59-68,1994; Stitt et al., Cell 80:661-670, 1995.
Anti-Inflammatory Activity Proteins of the present invention may also exhibit anti-inflammatory activity.
The anti-inflammatory activity may be achieved by providing a stimulus to cells involved in the inflammatory response, by inhibiting or promoting cell-cell interactions (such as, for example, cell adhesion), by inhibiting or promoting chemotaxis of cells involved in the inflammatory process, inhibiting or promoting cell extravasation, or by stimulating or suppressing production of other factors which more directly inhibit or promote an inflammatory response. Proteins exhibiting such activities can be used to treat inflammatory conditions including chronic or acute conditions), including without limitation inflammation associated with infection (such as septic shock, sepsis or systemic 2 0 inflammatory response syndrome (SIRS)), ischemia-reperfusion injury, endotoxin lethality, arthritis, complement-mediated hyperacute rejection, nephritis, cytokine or chemokine-induced lung injury, inflammatory bowel disease, Crohn's disease or resulting from over production of cytokines such as TNF or IL-1. Proteins of the invention may also be useful to treat anaphylaxis and hypersensitivity to an antigenic substance or material.
~adherin/Tumor Invasion Sup~essor Activit3r Cadherins are calcium-dependent adhesion molecules that appear to play major roles during development, particularly in defining specific cell types. Loss or alteration of normal cadherin expression can lead to changes in cell adhesion properties linked to 3 0 tumor growth and metastasis. Cadherin malfunction is also implicated in other human diseases, such as pemphigus vulgaris and pemphigus foliaceus (auto-immune blistering skin diseases), Crohn's disease, and some developmental abnormalities.
The cadherin superfamily includes well over forty members, each with a distinct pattern of expression. All members of the superfamily have in common conserved extracellular repeats (cadherin domains), but structural differences are found in other parts of the molecule. The cadherin domains bind calcium to form their tertiary structure and thus calcium is required to mediate their adhesion. Only a few amino acids in the first cadherin domain provide the basis for homophilic adhesion; modification of this recognition site can change the specificity of a cadherin so that instead of recognizing only itself, the mutant molecule can now also bind to a different cadherin. In addition, some cadherins engage in heterophilic adhesion with other cadherins.
E-cadherin, one member of the cadherin superfamily, is expressed in epithelial cell types. Pathologically, if E-cadherin expression is lost in a tumor, the malignant cells become invasive and the cancer metastasizes. Transfection of cancer cell lines with polynucleotides expressing E-cadherin has reversed cancer-associated changes by returning altered cell shapes to normal, restoring cells' adhesiveness to each other and to their substrate, decreasing the cell growth rate, and drastically reducing anchorage-independent cell growth. Thus, reintroducing E-cadherin expression reverts carcinomas to a less advanced stage. It is likely that other cadherins have the same invasion suppressor role in carcinomas derived from other tissue types. Therefore, proteins of the present invention with cadherin activity, and polynucleotides of the present invention encoding such proteins, can be used to treat cancer. Introducing such proteins or polynucleotides into cancer cells can reduce or eliminate the cancerous changes observed 2 0 in these cells by providing normal cadherin expression.
Cancer cells have also been shown to express cadherins of a different tissue type than their origin, thus allowing these cells to invade and metastasize in a different tissue in the body. Proteins of the present invention with cadherin activity, and polynucleotides of the present invention encoding such proteins, can be substituted in these cells for the 2 5 inappropriately expressed cadherins, restoring normal cell adhesive properties and reducing or eliminating the tendency of the cells to metastasize.
Additionally, proteins of the present invention with cadherin activity, and polynucleotides of the present invention encoding such proteins, can used to generate antibodies recognizing and binding to cadherins. Such antibodies can be used to block 3 0 the adhesion of inappropriately expressed tumor-cell cadherins, preventing the cells from forming a tumor elsewhere. Such an anti-cadherin antibody can also be used as a marker for the grade, pathological type, and prognosis of a cancer, i.e. the more progressed the cancer, the less cadherin expression there will be, and this decrease in cadherin expression can be detected by the use of a cadherin-binding antibody.

Fragments of proteins of the present invention with cadherin activity, preferably a polypeptide comprising a decapeptide of the cadherin recognition site, and poly-nucleotides of the present invention encoding such protein fragments, can also be used to block cadherin function by binding to cadherins and preventing them from binding in ways that produce undesirable effects. Additionally, fragments of proteins of the present invention with cadherin activity, preferably truncated soluble cadherin fragments which have been found to be stable in the circulation of cancer patients, and polynucleotides encoding such protein fragments, can be used to disturb proper cell-cell adhesion.
Assays for cadherin adhesive and invasive suppressor activity include, without limitation, those described in: Hortsch et al. J Biol Chem 270 (32): 18809-18817, 1995;
Miyaki et al. Oncogene 11: 2547-2552,1995; Ozawa et al. Cell 63: 1033-1038,1990.
Tumor Inhibition Activit3r In addition to the activities described above for immunological treatment or prevention of tumors, a protein of the invention may exhibit other anti-tumor activities.
A protein may inhibit tumor growth directly or indirectly (such as, for example, via antibody-dependent cell-mediated cytotoxicity (ADCC)). A protein may exhibit its tumor inhibitory activity by acting on tumor tissue or tumor precursor tissue, by inhibiting formation of tissues necessary to support tumor growth (such as, for example, by 2 0 inhibiting angiogenesis), by causing production of other factors, agents or cell types which inhibit tumor growth, or by suppressing, eliminating or inhibiting factors, agents or cell types which promote tumor growth.
Other Activities 2 5 A protein of the invention may also exhibit one or more of the following additional activities or effects: inhibiting the growth, infection or function of, or killing, infectious agents, including, without limitation, bacteria, viruses, fungi and other parasites; effecting (suppressing or enhancing) bodily characteristics, including, without limitation, height, weight, hair color, eye color, skin, fat to lean ratio or other tissue pigmentation, or organ 3 0 or body part size or shape (such as, for example, breast augmentation or diminution, change in bone form or shape); effecting biorhythms or caricadic cycles or rhythms;
effecting the fertility of male or female subjects; effecting the metabolism, catabolism, anabolism, processing, utilization, storage or elimination of dietary fat, lipid, protein, carbohydrate, vitamins, minerals, cofactors or other nutritional factors or component(s);

effecting behavioral characteristics, including, without limitation, appetite, libido, stress, cognition (including cognitive disorders), depression (including depressive disorders) and violent behaviors; providing analgesic effects or other pain reducing effects;
promoting differentiation and growth of embryonic stem cells in lineages other than hematopoietic lineages; hormonal or endocrine activity; in the case of enzymes, correcting deficiencies of the enzyme and treating deficiency-related diseases; treatment of hyperproliferative disorders (such as, for example, psoriasis); immunoglobulin-like activity (such as, for example, the ability to bind antigens or complement); and the ability to act as an antigen in a vaccine composition to raise an immune response against such protein or another material or entity which is cross-reactive with such protein.
ADMINISTRATION AND DOSING
A protein of the present invention (from whatever source derived, including without limitation .from recombinant and non-recombinant sources) may be used in a pharmaceutical composition when combined with a pharmaceutically acceptable Garner.
Such a composition may also contain (in addition to protein and a carrier) diluents, fillers, salts, buffers, stabilizers, solubilizers, and other materials well known in the art. The term "pharmaceutically acceptable' means a non-toxic material that does not interfere with the effectiveness of the biological activity of the active ingredient(s). The characteristics of the 2 0 carrier will depend on the route of administration. The pharmaceutical composition of the invention may also contain cytokines, lymphokines, or other hematopoietic factors such as M-CSF, GM-CSF, TNF, IL-1, IL-2, IL-3, IL-4, IL-5, IL-6, IL-7, ILr8, IL-9, IL-10, IL-11, IL-12, IL-13, IL-14, IL-15, IFN, TNFO, TNF1, TNF2, G-CSF, Meg-CSF, thrombopoietin, stem cell factor, and erythropoietin. The pharmaceutical composition may further contain other 2 5 agents which either enhance the activity of the protein or compliment its activity or use in treatment. Such additional factors and/or agents may be included in the pharmaceutical composition to produce a synergistic effect with protein of the invention, or to minimize side effects. Conversely, protein of the present invention may be included in formulations of the particular cytokine, lymphokine, other hematopoietic factor, 3 0 thrombolytic or anti-thrombotic factor, or anti-inflammatory agent to minimize side effects of the cytokine, lymphokine, other hematopoietic factor, thrombolytic or anti-thrombotic factor, or anti-inflammatory agent.
A protein of the present invention may be active in multimers (e.g., heterodimers or homodimers) or complexes with itself or other proteins. As a result, pharmaceutical compositions of the invention may comprise a protein of the invention in such multimeric or complexed form.
The pharmaceutical composition of the invention may be in the form of a complex of the proteins) of present invention along with protein or peptide antigens.
The protein and/or peptide antigen will deliver a stimulatory signal to both B and T
lymphocytes. B
lymphocytes will respond to antigen through their surface immunoglobulin receptor. T
lymphocytes will respond to antigen through the T cell receptor (TCR) following presentation of the antigen by MHC proteins. MHC and structurally related proteins including those encoded by class I and class II MHC genes on host cells will serve to present the peptide antigens) to T lymphocytes. The antigen components could also be supplied as purified MHC-peptide complexes alone or with co-stimulatory molecules that can directly signal T cells. Alternatively antibodies able to bind surface immunolgobulin and other molecules on B cells as well as antibodies able to bind the TCR and other molecules on T cells can be combined with the pharmaceutical composition of the invention.
The pharmaceutical composition of the invention may be in the form of a liposome in which protein of the present invention is combined, in addition to other pharmaceutically acceptable carriers, with amphipathic agents such as lipids which exist in aggregated form as micelles, insoluble monolayers, liquid crystals, or lamellar layers 2 0 in aqueous solution. Suitable lipids for liposomal formulation include, without limitation, monoglycerides, diglycerides, sulfatides, lysolecithin, phospholipids, saponin, bile acids, and the like. Preparation of such liposomal formulations is within the level of skill in the art, as disclosed, for example, in U.S. Patent No. 4,235,871; U.S. Patent No.
4,502,728; U.S.
Patent No. 4,837,028; and U.S. Patent No. 4,737,323, all of which are incorporated herein 2 5 by reference.
As used herein, the term "therapeutically effective amount" means the total amount of each active component of the pharmaceutical composition or method that is sufficient to show a meaningful patient benefit, i.e., treatment, healing, prevention or amelioration of the relevant medical condition, or an increase in rate of treatment, healing, 3 0 prevention or amelioration of such conditions. When applied to an individual active ingredient, administered alone, the term refers to that ingredient alone. When applied to a combination, the term refers to combined amounts of the active ingredients that result in the therapeutic effect, whether administered in combination, serially or simultaneously.

In practicing the method of treatment or use of the present invention, a therapeutically effective amount of protein of the present invention is administered to a mammal having a condition to be treated. Protein of the present invention may be administered in accordance with the method of the invention either alone or in combination with other therapies such as treatments employing cytolcines, lympholcines or other hematopoietic factors. When co-administered with one or more cytokines, lympholcines or other hematopoietic factors, protein of the present invention may be administered either simultaneously with the cytolcine(s), lymphokine(s), other hematopoietic factor(s), thrombolytic or anti-thrombotic factors, or sequentially. If administered sequentially, the attending physician will decide on the appropriate sequence of administering protein of the present invention in combination with cytolcine(s), lympholcine(s), other hematopoietic factor(s), thrombolytic or anti-thrombotic factors.
Administration of protein of the present invention used in the pharmaceutical composition or to practice the method of the present invention can be carried out in a variety of conventional ways, such as oral ingestion, inhalation, topical application or cutaneous, subcutaneous, intraperitoneal, parenteral or intravenous injection.
Intravenous administration to the patient is preferred.
When a therapeutically effective amount of protein of the present invention is 2 0 administered orally, protein of the present invention will be in the form of a tablet, capsule, powder, solution or elixir. When administered in tablet form, the pharmaceutical composition of the invention may additionally contain a solid carrier such as a gelatin or an adjuvant. The tablet, capsule, and powder contain from about 5 to 95%
protein of the present invention, and preferably from about 25 to 90% protein of the present invention.
2 5 When administered in liquid form, a liquid Garner such as water, petroleum, oils of animal or plant origin such as peanut oil, mineral oil, soybean oil, or sesame oil, or synthetic oils may be added. The liquid form of the pharmaceutical composition may further contain physiological saline solution, dextrose or other saccharide solution, or glycols such as ethylene glycol, propylene glycol or polyethylene glycol. When administered in liquid 3 0 form, the pharmaceutical composition contains from about 0.5 to 90% by weight of protein of the present invention, and preferably from about 1 to 50% protein of the present invention.
When a therapeutically effective amount of protein of the present invention is administered by intravenous, cutaneous or subcutaneous injection, protein of the present invention will be in the form of a pyrogen-free, parenterally acceptable aqueous solution.
The preparation of such parenterally acceptable protein solutions, having due regard to pH, isotonicity, stability, and the like, is within the skill in the art. A
preferred pharmaceutical composition for intravenous, cutaneous, or subcutaneous injection should contain, in addition to protein of the present invention, an isotonic vehicle such as Sodium Chloride Injection, Ringer's Injection, Dextrose Injection, Dextrose and Sodium Chloride Injection, Lactated Ringei s Injection, or other vehicle as known in the art.
The pharmaceutical composition of the present invention may also contain stabilizers, preservatives, buffers, antioxidants, or other additives known to those of skill in the art.
The amount of protein of the present invention in the pharmaceutical composition of the present invention will depend upon the nature and severity of the condition being treated, and on the nature of prior treatments which the patient has undergone.
Ultimately, the attending physician will decide the amount of protein of the present invention with which to treat each individual patient. Initially, the attending physician will administer low doses of protein of the present invention and observe the patient's response. Larger doses of protein of the present invention may be administered until the optimal therapeutic effect is obtained for the patient, and at that point the dosage is not increased further. It is contemplated that the various pharmaceutical compositions used to practice the method of the present invention should contain about 0.01 izg to about 100 2 0 mg (preferably about O.lng to about 10 mg, more preferably about 0.1 izg to about 1 mg) of protein of the present invention per kg body weight.
The duration of intravenous therapy using the pharmaceutical composition of the present invention will vary, depending on the severity of the disease being treated and the condition and potential idiosyncratic response of each individual patient.
It is 2 5 contemplated that the duration of each application of the protein of the present invention will be in the range of 12 to 24 hours of continuous intravenous administration.
Ultimately the attending physician will decide on the appropriate duration of intravenous therapy using the pharmaceutical composition of the present invention.
Protein of the invention may also be used to immunize animals to obtain 3 0 polyclonal and monoclonal antibodies which specifically react with the protein. As used herein, the term "antibody" includes without limitation a polyclonal antibody, a monoclonal antibody, a chimeric antibody, a single-chain antibody, a CDR-grafted antibody, a humanized antibody, or fragments thereof which bind to the indicated protein.

Such term also includes any other species derived from an antibody or antibody sequence which is capable of binding the indicated protein.
Antibodies to a particular protein can be produced by methods well known to those skilled in the art. For example, monoclonal antibodies can be produced by generation of antibody-producing hybridomas in accordance with known methods (see for example, Goding, 1983, Monoclonal antibodies: principles and practice, Academic Press Inc., New York; and Yokoyama,1992, "Production of Monoclonal Antibodies" in Current Protocols in Immunology, Unit 2.5, Greene Publishing Assoc. and John Wiley & Sons).
Polyclonal sera and antibodies can be produced by inoculation of a mammalian subject with the relevant protein or fragments thereof in accordance with known methods.
Fragments of antibodies, receptors, or other reactive peptides can be produced from the corresponding antibodies by cleavage of and collection of the desired fragments in accordance with known methods (see for examgle, Goding, supra; and Andrew et al., 1992, "Fragmentation of Immunoglobulins" in Current Protocols in Immunology, Unit 2.8, Greene Publishing Assoc. and John Wiley & Sons). Chimeric antibodies and single chain antibodies can also be produced in accordance with known recombinant methods (see for example, 5,169,939, 5,194,594, and 5,576,184). Humanized antibodies can also be made from corresponding marine antibodies in accordance with well known methods (see for example, U.S.
Patent Nos. 5,530,101, 5,585,089, and 5,693,762). Additionally, human antibodies may be 2 0 produced in non-human animals such as mice that have been genetically altered to express human antibody molecules (see for example Fishwild et al., 1996, Nature Biotechnology 14: 845-851; Mendez et al., 1997, Nature Genetics 15: 146-156 (erratum Nature Genetics 16: 410); and U.S. Patents 5,877,397 and 5,625,126). Such antibodies may be obtained .
using either the entire protein or fragments thereof as an immunogen. The peptide 2 5 immunogens additionally may contain a cysteine residue at the carboxyl terminus, and are conjugated to a hapten such as keyhole limpet hemocyanin (KLH). Methods for synthesizing such peptides are known in the art, for example, as in R.P.
Merrifield, J.
Amer.Chem.Soc. ~,5, 2149-2154 (1963); J.L. Krstenansky, et al., FEBS Lett.
211,10 (198.
Monoclonal antibodies binding to the protein of the invention may be useful 3 0 diagnostic agents for the immunodetection of the protein. Neutralizing monoclonal antibodies binding to the protein may also be useful therapeutics for both conditions associated with the protein and also in the treatment of some forms of cancer where abnormal expression of the protein is involved. In the case of cancerous cells or leukemic cells, neutralizing monoclonal antibodies against the protein may be useful in detecting and preventing the metastatic spread of the cancerous cells, which may be mediated by the protein.
For compositions of the present invention which are useful for bone, cartilage, tendon or ligament regeneration, the therapeutic method includes administering the composition topically, systematically, or locally as an implant or device.
When administered, the therapeutic composition for use in this invention is, of course, in a pyrogen-free, physiologically acceptable form. Further, the composition may desirably be encapsulated or injected in a viscous form for delivery to the site of bone, cartilage or tissue damage. Topical administration may be suitable for wound healing and tissue repair. Therapeutically useful agents other than a protein of the invention which may also optionally be included in the composition as described above, may alternatively or additionally, be administered simultaneously or sequentially with the composition in the methods of the invention. Preferably for bone and/or cartilage formation, the composition would include a matrix capable of delivering the protein-containing composition to the site of bone and/or cartilage damage, providing a structure for the developing bone and cartilage and optimally capable of being resorbed into the body.
Such matrices may be formed of materials presently in use for other implanted medical 2 0 applications.
The choice of matrix material is based on biocompatibility, biodegradability, mechanical properties, cosmetic appearance and interface properties. The particular application of the compositions will define the appropriate formulation.
Potential matrices for the compositions may be biodegradable and chemically defined calcium 2 5 sulfate, tricalciumphosphate, hydroxyapatite, polylactic acid, polyglycolic acid and polyanhydrides. Other potential materials are biodegradable and biologically well-defined, such as bone or dermal collagen. Further matrices are comprised of pure proteins or extracellular matrix components. Other potential matrices are nonbiodegradable and chemically defined, such as sintered hydroxapatite, bioglass, aluminates, or other 3 0 ceramics. Matrices may be comprised of combinations of any of the above mentioned types of material, such as polylactic acid and hydroxyapatite or collagen and tricalciumphosphate. The bioceramics may be altered in composition, such as in calcium-aluminate-phosphate and processing to alter pore size, particle size, particle shape, and biodegradability.

Presently preferred is a 50:50 (mole weight) copolymer of lactic acid and glycolic acid in the form of porous particles having diameters ranging from 150 to 800 microns.
In some applications, it will be useful to utilize a sequestering agent, such as carboxymethyl cellulose or autologous blood clot, to prevent the protein compositions from disassociating from the matrix.
A preferred family of sequestering agents is cellulosic materials such as alkylcelluloses (including hydroxyalkylcelluloses), including methylcellulose, ethylcellulose, hydroxyethylcellulose, hydroxypropylcellulose, hydroxypropyl-methylcellulose, and carboxymethylcellulose, the most preferred being cationic salts of carboxymethylcellulose (CMC). Other preferred sequestering agents include hyaluronic acid, sodium alginate, polyethylene glycol), polyoxyethylene oxide, carboxyvinyl polymer and polyvinyl alcohol). The amount of sequestering agent useful herein is 0.5-20 wt%, preferably 1-10 wt% based on total formulation weight, which represents the amount necessary to prevent desorbtion of the protein from the polymer matrix and to provide appropriate handling of the composition, yet not so much that the progenitor cells are prevented from infiltrating the matrix, thereby providing the protein the opportunity to assist the osteogenic activity of the progenitor cells.
In further compositions, proteins of the invention may be combined with other agents beneficial to the treatment of the bone and/or cartilage defect, wound, or tissue in 2 0 question. These agents include various growth factors such as epidermal growth factor (EGF), platelet derived growth factor (PDGF), transforming growth factors (TGF-a and TGF-Vii), and insulin-like growth factor (IGF).
The therapeutic compositions are also presently valuable for veterinary applications. Particularly domestic animals and thoroughbred horses, in addition to 2 5 humans, are desired patients for such treatment with proteins of the present invention.
The dosage regimen of a protein-containing pharmaceutical composition to be used in tissue regeneration will be determined by the attending physician considering various factors which modify the action of the proteins, e.g., amount of tissue weight desired to be formed, the site of damage, the condition of the damaged tissue, the size of 3 0 a wound, type of damaged tissue (e.g., bone), the patient's age, sex, and diet, the severity of any infection, time of administration and other clinical factors. The dosage may vary with the type of matrix used in the reconstitution and with inclusion of other proteins in the pharmaceutical composition. For example, the addition of other known growth factors, such as IGF I (insulin like growth factor I), to the final composition, may also effect the dosage. Progress can be monitored by periodic assessment of tissue/bone growth and/or repair, for example, X-rays, histomorphometric determinations and tetracycline labeling.
Polynucleotides of the present invention can also be used for gene therapy.
Such polynucleotides can be introduced either in vivo or ex vivo into cells for expression in a mammalian subject. Polynucleotides of the invention may also be administered by other known methods for introduction of nucleic acid into a cell or organism (including, without limitation, in the form of viral vectors or naked DNA).
Cells may also be cultured ex vivo in the presence of proteins of the present invention in order to proliferate or to produce a desired effect on or activity in such cells.
Treated cells can then be introduced in vivo for therapeutic purposes.
Patent and literature references cited herein are incorporated by reference as if fully set forth.

SEQUENCE LISTING
<I10> Jacobs, Kenneth McCoy, John M.
LaVallie, Edward R.
Collins-Racie, Lisa A.
Evans, Cheryl Merberg, David Treacy, Maurice Agostino, Michael J.
Steininger II, Robert J.
Genetics Institute, Inc.
<120> SECRETED PROTEINS AND POLYNUCLEOTIDES ENCODING THEM
<130> GI 6067A
<140>
<141>
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<221> unsure <222> (2343) <400> 1 ccacttcccc cttttgttaa ttaaaactaa gaagtcggaa tgggaacgag gtgcccagct 60 cccgtggaga aagcttaagg acaccacgcc agtgctttcc tgccttcctt ccgagatgga 120 aagaggagct cctagctcac ttaagccggg gtagggctgg ttctcctttc cgagccaaaa 180 tcccaggcga tggtgaatta tgaacgtgcc acaccatgaa gctcttgtgg caggtaactg 240 tgcaccacca cacctggaat gccatcctgc tcccgttcgt ctacctcacg gcgcaagtgt 300 ggattctgtg tgcagccatc gctgctgccg cctcagccgg gccccagaac tgcccctccg 360 tctgctcgtg cagtaaccag ttcagcaagg tggtgtgcac gcgccggggc ctctccgagg 420 tcccgcaggg tattccctcg aacacccggt acctcaacct catggagaac aacatccaga 480 tgatccaggc cgacaccttc cgccacctcc accacctgga ggtcctgcag ttgggcagga 540 actccatccg gcagattgag gtgggggcct tcaacggcct ggccagcctc aacaccctgg 600 agctgttcga caactggctg acagtcatcc ctagcggggc ctttgaatac ctgtccaagc 660 tgcgggagct ctggcttcgc aacaacccca tcgaaagcat cccctcttac gccttcaacc 720 gggtgccctc cctcatgcgc ctggacttgg gggagctcaa gaagctggag tatatctctg 780 agggagcttt tgaggggctg ttcaacctca agtatctgaa cttgggcatg tgcaacatta 840 aagacatgcc caatctcacc cccctggtgg ggctggagga gctggagatg tcagggaacc 900 acttccctga gatcaggcct ggctccttcc atggcctgag ctccctcaag aagctctggg 960 tcatgaactc acaggtcagc ctgattgagc ggaatgcttt tgacgggctg gcttcacttg 1020 tggaactcaa cttggcccac aataacctct cttctttgcc ccatgacctc tttaccccgc 1080 tgaggtacct ggtggagttg catctacacc acaacccttg gaactgtgat tgtgacattc 1140 tgtggctagc ctggtggctt cgagagtata tacccaccaa ttccacctgc tgtggccgct 1200 gtcatgctcc catgcacatg cgaggccgct acctcgtgga ggtggaccag gcctccttcc 1260 agtgctctgc ccccttcatc atggacgcac ctcgagacct caacatttct gagggtcgga 1320 tggcagaact taagtgtcgg actcccccta tgtcctccgt gaagtggttg ctgcccaatg 1380 ggacagtgct cagccacgcc tcccgccacc caaggatctc tgtcctcaac gacggcacct 1440 tgaacttttc ccacgtgctg ctttcagaca ctggggtgta cacatgcatg gtgaccaatg 1500 ttgcaggcaa ctccaacgcc tcggcctacc tcaatgtgag cacggctgag cttaacacct 1560 ccaactacag cttcttcacc acagtaacag tggagaccac ggagatctcg cctgaggaca 1620 caacgcgaaa gtacaagcct gttcctacca cgtccactgg ttaccagccg gcatatacca 1680 cctctaccac ggtgctcatt cagactaccc gtgtgcccaa gcaggtggca gtacccgcga 1740 cagacaccac tgacaagatg cagaccagcc tggatgaagt catgaagacc accaagatca 1800 tcattggctg ctttgtggca gtgattgtgc tagctgccgc catgttgatt gtcttctata 1860 aacttcgtaa gcggcaccag cagcggagta cagtcacagc cgcccggact gttgagataa 1920 tccaggtgga cgaagacatc ccagcagcaa catccgcagc agcaacagca gctccgtccg 1980 gtgtatcagg tgagggggca gtagtgctgc ccacaattca tgaccatatt aactacaaca 2040 cctacaaacc agcacatggg gcceactgga cagaaaacag cctggggaac tctctgcacc 2100 ccacagtcac cactatctct gaaccttata taattcagac ccataccaag gacaaggtac 2160 aggaaactca aatatgactc cctcccccaa aaaacttata aaatgcaata gsatgcacac 2220 aaagacagca acttttgtac agagtgggga gagacttttt cttgtatatg cttatatatt 2280 aagtctatgg gctggttaaa aaaaacagat tatattaaaa tttaaagaca aaaagtcaaa 2340 acnaaaaaaa aaaaaaaaaa 2360 <210> 2 <211> 653 <212> PRT
<213> Homo Sapiens <400> 2 Met Lys Leu Leu Trp Gln Val Thr Val His His His Thr Trp Asn Ala Ile Leu Leu Pro Phe Val Tyr Leu Thr Ala Gln Val Trp Ile Leu Cys Ala Ala Ile Ala Ala Ala Ala Ser Ala Gly Pro Gln Asn Cys Pro Ser Val Cys Ser Cys Ser Asn Gln Phe Ser Lys Val Val Cys Thr Arg Arg Gly Leu Ser Glu Val Pro Gln Gly Ile Pro Ser Asn Thr Arg Tyr Leu Asn Leu Met Glu Asn Asn Ile Gln Met Ile Gln Ala Asp Thr Phe Arg His Leu His His Leu Glu Val Leu Gln Leu Gly Arg Asn Ser Ile Arg Gln Ile Glu Val Gly Ala Phe Asn Gly Leu Ala Ser Leu Asn Thr Leu Glu Leu Phe Asp Asn Trp Leu Thr Val Ile Pro Ser Gly Ala Phe Glu Tyr Leu Ser Lys Leu Arg Glu Leu Trp Leu Arg Asn Asn Pro Ile Glu Ser Ile Pro Ser Tyr Ala Phe Asn Arg Val Pro Ser Leu Met Arg Leu Asp Leu Gly Glu Leu Lys Lys Leu Glu Tyr Ile Ser Glu Gly Ala Phe Glu Gly Leu Phe Asn Leu Lys Tyr Leu Asn Leu Gly Met Cys Asn Ile Lys Asp Met Pro Asn Leu Thr Pro Leu Val Gly Leu Glu Glu Leu Glu Met Ser Gly Asn His Phe Pro Glu Ile Arg Pro Gly Ser Phe His Gly Leu Ser Ser Leu Lys Lys Leu Trp Val Met Asn Ser Gln Val Ser Leu Ile Glu Arg Asn Ala Phe Asp Gly Leu Ala Ser Leu Val Glu Leu Asn Leu Ala His Asn Asn Leu Ser Ser Leu Pro His Asp Leu Phe Thr Pro Leu Arg Tyr Leu Val Glu Leu His Leu His His Asn Pro Trp Asn Cys Asp Cys Asp Ile Leu Trp Leu Ala Trp Trp Leu Arg Glu Tyr Ile Pro Thr Asn Ser Thr Cys Cys Gly Arg Cys His Ala Pro Met His Met Arg Gly Arg Tyr Leu Val Glu Val Asp Gln Ala Ser Phe Gln Cys Ser Ala Pro Phe Ile Met Asp Ala Pro Arg Asp Leu Asn Ile Ser Glu Gly Arg Met Ala Glu Leu Lys Cys Arg Thr Pro Pro Met Ser Ser Val Lys Trp Leu Leu Pro Asn Gly Thr Val Leu Ser His Ala Ser Arg His Pro Arg Ile Ser Val Leu Asn Asp Gly Thr Leu Asn Phe Ser His Val Leu Leu Ser Asp Thr Gly Val Tyr Thr Cys Met Val Thr Asn Val Ala Gly Asn Ser Asn Ala Ser Ala Tyr Leu Asn Val Ser Thr Ala Glu Leu Asn Thr Ser Asn Tyr Ser Phe Phe Thr Thr Val Thr Val Glu Thr Thr Glu Ile Ser Pro Glu Asp Thr Thr Arg Lys Tyr Lys Pro Val Pro Thr Thr Ser Thr Gly Tyr Gln Pro Ala Tyr Thr Thr Ser Thr Thr Val Leu Ile Gln Thr Thr Arg Val Pro Lys Gln Val Ala Val Pro Ala Thr Asp Thr Thr Asp Lys Met Gln Thr Ser Leu Asp Glu Val Met Lys Thr Thr Lys Ile WO 99!50405 PCTNS99/06946 Ile Ile Gly Cys Phe Val Ala Val Ile Val Leu Ala Ala Ala Met Leu Ile Val Phe Tyr Lys Leu Arg Lys Arg His Gln Gln Arg Ser Thr Val Thr Ala Ala Rrg Thr Val Glu Ile Ile Gln Val Asp Glu Asp Ile Pro Ala Ala Thr Ser Ala Ala Ala Thr Ala Ala Pro Ser Gly Val Ser Gly Glu Gly Ala Val Val Leu Pro Thr Ile His Asp His Ile Asn Tyr Asn Thr Tyr Lys Pro Ala His Gly Ala His Trp Thr Glu Asn Ser Leu Gly Asn Ser Leu His Pro Thr Val Thr Thr Ile Ser Glu Pro Tyr Ile Ile Gln Thr His Thr Lys Asp Lys Val Gln Glu Thr Gln Ile <210> 3 <211> 1558 <212> DNA
<213> Homo Sapiens <400> 3 gcgaagctga taagccaata agagtgctgt ctagaacgca tttgcaaacc gttaagagta 60 gcttcttttt gaggcctgga cttctgattc atctatagaa gtctattatg attccatcaa 120 ttgcatagct tgtgtacctg gtagaaattg tgtcttggaa tgaccctttc gagttattga 180 catggctctg atgaatagaa catgagcccc aaaactaaat ccaaaaggaa ttttctatct 240 ttcatccccc acatgtggca agacaagttg gccctttctt acccagaggt cttttgtgtg 300 actgcatctt tctcctccgt tctccattgt gtgctttcca ttttgtcttt agtgcctata 360 ctgttaggtg ttttcttcac tggcattcac aaatttaagc cattgctgcc tcattagcct 420 tgtattttgt gtgcatatca tatatccaga cctgtatgtt cactttaagc attcttatat 480 cacactgtct cctcatctac catatggtaa atgaggagac agtgtcttat atcacactgt 540 ctcctcatct accatatggt aaatgttaaa actccacatt tgtctgcatc agggaaaatg 600 catgggcaca catcctccct ccctccctct ctgctctcct cccttccttc aggcctctta 660 gcattgtttg ttttcccatt tctgatacta ctactccatg ctgaagattt gccatattac 720 tattttggaa acattgagtg atagaactcc tagaaaattt gcaaagaaat gttacatact 780 gtatatcaaa ctctcagatt ctagtgttga aaaagtagcc tatactttgc tattacttat 840 acctgctgcc atagaaaaaa ataagtttat tcatgacaca tttacatttg atcataaata 900 aaagaaaaaa gggcaccttt ttggagttag tcatggtagt cattagtgat atttctgaac 960 agttcttaat ttaaaatact tcaaaggaag taaaggtcat ggcttagctg aaggaaatgc 1020 tccagaaatt ggactgtgta aaccatcagt acaataatac gctgtgtatg tatgtgtata 1080 taaaatgaga attatggcat aattggagca tttgcattaa tcaacaaact cacattgaga 1140 caaaacttag ttttacagct gtcttgatta aagccaagtg ttccatgttg ctgtgaagaa 1200 tagcctcttt caaatacttt ggaaagtagt tacttggaaa cttgtaaagg tattacattt 1260 ttatatttaa acacctatag agatcttcaa ttccttgagt ctgagcttgt gggtggaatt 1320 ctaaatttgt atcataatct gtcttttgtg aaacattttg aaaatatgta tatataatat 1380 tgtatatgca aattgtgttg tttcacttgt aaagggaaaa ggcttatttt tctttatatt 1440 tctgataact tgttttgcat atgaccagca ctgactgaaa ggcatgtgta gctgcaaaca 1500 ctgttgcttt ttttgtgaaa tgaaaataaa agtatttaaa tacaaaaaaa aaaaaaaa 1558 <210> 4 <211> 71 <212> PRT
<213> Homo sapiens <400> 4 Met Ser Pro Lys Thr Lys Ser Lys Arg Asn Phe Leu Ser Phe Ile Pro His Met Trp Gln Asp Lys Leu Ala Leu Ser Tyr Pro Glu Val Phe Cys Val Thr Ala Ser Phe Ser Ser Val Leu His Cys Val Leu Ser Ile Leu Ser Leu Val Pro Ile Leu Leu Gly Val Phe Phe Thr Gly Ile His Lys Phe Lys Pro Leu Leu Pro His <210> 5 <211> 2637 <212> DNA
<213> Homo Sapiens <400> 5 ccttgagcct gttcctttac ctcaccaggc ctcattttcc acatctgttg aacaatctgg 60 ctcttagggc ccctttcaat tctagtgttc tggaggtcca tgaactgttt ttgtgtagta 120 gagaagaact ttcctgaagt tcgaatatat aatatatgac agaggaastc actgttatga 180 accttagagt tattcccatg ggaggatgaa gactgacatt gcttccttgc acaccaggac 240 ttgggcctgt actgctctaa ttctgaactt aatttcctqt gtggctcttg cttggttcat 300 gggacggtct cctaacataa tgtttatatc ccgtggctca ggcatagaga cccactcatg 360 gtttcccatc agaatttggg ttccagttca ccactccaca tacacacacc aaggctaatg 420 attcatgcta ttgtcagttt ttgcttcttt gatattgttc tgaaggtagt ttttattgat 480 acttctttct ttctaaagtc aatgagtaaa gttatggcaa gacattggaa agacattgca 540 gtgctcccag gctggagtgc atgactcatc tcctggcccc tccaggaaac cggagaaggc 600 agaaaagggc aagaatgttg tgttgtcttt gggctgtgga gttcatgatt atggacactt 660 tctgaattat tggcactaag ggcacatcct gcttagcatg gaggctggac agctctggga 720 aagcagagat tctttttagg gagtccagag catttttttc ttctttctta tcccctcttt 780 ccaatagaga tgtcagaagg gtgactgtta gggttgatac attatgtggt gctctgtttt 840 tttgagcatt gtctactcag agtgattacc aaggtggagc tcattatgtt acctggcacc 900 tgcaactact tgcctcatgt tggactaaag tggaaagact cctattggga attcactcag 960 ggaggcacca ggcattgtta gaaaagcagt atcgcagagc acttaagagc tgagccttgt 1020 gatcgggtgg acttgggttt atgtcctcgc actgtcgctt cctagattta tgacctcagt 1080 ttcctcattt ctaaagtaga gacagggatg gagggtgggg agaatatgaa ttaatttgga 1140 ttaaatggat gagaatcaga agtatttatt tggaattggt gtgaattgga gcacatgggg 1200 aactgaaaac aaaggtgata atttatgtgg tgtagatgaa ctaagaataa tgaaaacagc 1260 atgaaggatg gaatcaaatt aagtgcaaac atataaagga aatggactgg taaaagctcg 1320 agggcaaagg tgtatggaga tgggaattca ggttggaaag aggctgctga tacaaggact 1380 ggagttggga gagctgctga ccaaatgtgg aaatgaagat tttttaaaaa tgggcgctgg 1440 ttggattcag gcaagctatt ttaacccagg actcacttta attcagtcaa aatatttctt 1500 atcttccagt tggggaggtg ttttgggaca ggaaaaaaag ggagataaga ataccatccc 1560 agagaaagaa gagaagtatt tgcattttgt tctctagagt tgcagatata tgctgtatct 1620 cttctatatg gtaatacatt gcaaaggaga tacagtgaag aaatgaagta aaataaaatg 1680 tgtgtgtctt cctctcatat gcataaagga agacaataca tgttggtaac atggatttgc 1740 ttgcattgaa gaatttttaa ggtgttattc atttaatctt tatatagaca gaatttactg 1800 agctgaaaaa gatatgcatt tttttccttc tgtggctgtt ttttgaggat gctgaatcct 1860 tcaaaatttt tgccctggag gatattaggc aatgaggtga ttttttattt attatttttt 1920 aaggtagcag atgttcacat tggtagctag ggcacaaacg caggttgccc aaggaagtaa 1980 agttagagat gggggtactg agctggaggt gcaggataac gaggcgagga atgggacaaa 2040 WO 99!50405 PCT/US99/06946 gggaatatta tgaagtagca ggatttagtg aaaggaagca tataaaaatg gacataaggg 2100 ccaggtgcgg tggctcacgc ccctaatcct agcactttgg gaggcctagg caggtggatc 2160 acctaaggtc aggagttcga gaccagcctg accaacaccc gtctctacta aaagtacaaa 2220 aattagctgg gcgtgtggcg cgtgcctgta gtcccagcta ctcaggaggc tgaggcagga 2280 gaatcgcttg accccaggag gtggaggttg cagtgcgccg agattgcgtc actgcactcc 2340 agcctgggtg acagagtgag acaccgtctc aaaaacaata aaaattaaaa aaaatggaca 2400 tatggtttat tcattgggaa gagcacctag tgttggcatg agtgtaaaaa tgttcttagc 2460 agccaagtca ttgcagacct ttctctccta ctmtgtgttc tggtcttcat atatttttct 2520 ttctcacaaa ttctaccatt atgatcatag ttgtattcat cttcttgagt cagctaaata 2580 atccttatat tttgatgaaa ggtaaccttc acttccaata ttaaaaaaaa aaaaaaa 2637 <210> 6 <211> 70 <212> PRT
<213> Homo sapiens <400> 6 Met Lys Thr Asp Ile Ala Ser Leu His Thr Arg Thr Trp Ala Cys Thr Ala Leu Ile Leu Asn Leu Ile Ser Cys Val Ala Leu Ala Trp Phe Met Gly Arg Ser Pro Asn Ile Met Phe Ile Ser Arg Gly Ser Gly Ile Glu Thr His Ser Trp Phe Pro Ile Arg Ile Trp Val Pro Val His His Ser Thr Tyr Thr His Gln Gly <210> 7 <211> 4398 <212> DNA
<213> Homo sapiens <220>
<221> unsure <222> (4338) <400> 7 ggcgctggtg gctgcggcgg cggcggcggc agcggcgctc gagcggttcc tgtcagggtc 60 agccggcggg ccccctgggt ggtccacctg caaatcgcgg agcggcgccc cagggatcga 120 tggcgatgaa ctataacgcg aaggatgaag tggacggtgg gcccccgtgt gctccggggg 180 gcaccgcgaa gactcggaga ccggataaca cggccttcaa acagcaacgg ctgccagctt 240 ggcagcccat ccttacggct ggcacggtgc tacctatttt cttcatcatc ggtctcatct 300 tcattcccat cggcattggc atttttgtca cctccaacaa catccgcgag atcgagattg 360 attataccgg aacagagcct tccagtccct gtaataaatg tttatctccg gatgtgacac 420 cttgcttttg taccattaac ttcacactgg aaaagtcatt tgagggcaac gtgtttatgt 480 attatggact gtctaatttc tatcaaaacc atcgtcgtta cgtgaaatct cgagatgata 540 gtcaactaaa tggagattct agtgctttgc ttaatcccag taaggaatgt gaaccttatc 600 gaagaaatga agacaaacca attgctcctt gtggagctat tgccaacagc atgtttaatg 660 atacattaga attgtttctc attggcaatg attcttatcc tatacctatc gctttgaaaa 720 agaaaggtat tgcttggtgg acagataaaa atgtgaaatt cagaaatccc cctggaggag 780 acaacctgga agaacgattt aaaggtacaa caaagcctgt gaactggctt aaaccagttt 840 acatgctgga ttctgaccca gataataatg gattcataaa tgaggatttt attgtttgga 900 tgcgtactgc agcattacct acttttcgca agttgtatcg tcttatagaa aggaaaagtg 960 atttacatcc aacattacca gctggccgat actctttgaa tgtcacatac aattaccctg 1020 tacattattt tgatggacga aaacggatga tcttgagcac tatttcatgg atgggaggaa 1080 aaaatccatt tttggggatt gcttacatcg ctgttggatc catctccttc cttctgggag 1140 ttgtactgct agtaattaat cataaatata gaaacagtag taatacagct gacattacca 1200 tttaatttta tattatgaaa gcaaatcatc tgcatgtgca tcaaggccag tcctattcaa 1260 cctagctttc gaatgctgat atctggttag tatgtcattt tgaagttggc acataacttt 1320 tctaaaaaaa agcagtcttt gttgtttgct tcttccctac ggatgacttc taaaaatata 1380 tgacgggtat aaaaaaatta gctatattga tcatatcaac actgtaactg ctgaaatggc 1440 attctaatgt ttgcttttta ttcggacagg ccacatgatg catagagcct ctttcatgtg 1500 acttgtgtct actgcttaaa tctttatgct gtgttgatga tattatattg acatatgaag 1560 ctgtatatgt gtatgtattt tgtggagaag agggattaca agatgtatga gtataatgac 1620 ttgctaacct ttcaggattc cgagaaagat gaagaaagac catatctaaa taatacactt 1680 catcattttc atgtgtataa atgcttaaag taccatcttt gttgaggtgg ttcatgtatc 1740 cagtttatcc agtacagtta tttgtcaagc ttagctttga tttcaaagga cacgcttacc 1800 ttgtctggca taagaattaa tgctcatgtc tgcagtggtt gggtaggtcc tgcttaggag 1860 aattaaaaaa ttcctctttc cgtttggttg aatgttgcag tcaggaaccc caactcactt 1920 ggaatgtttt tatatgtaat catttccctt gaagcttata ctttataagg gaagaaagaa 1980 ttcaggtgat atgggaaaac tgcttggcag accttcatct tctgcctcaa ctgtsaacca 2040 catgtaaatg cttaatggag actgttttca ttcttgtgat atttaacatt cagaaaatta 2100 cttcagcttt ggaaatactc aggctgtttt tattctgcag gtaagtgttt tgacttaagt 2160 actaatattc cagaaatttt tgaaagcagt aaccttaatt tcctatgtat ttcattccac 2220 ttttgcatat aggtcaaata gcaatgtgta tgcacattct ctttagttaa ggcaccaatt 2280 gttttggttg gttttcctaa gacatacttt aaaaagatgt tctataaatt tcctagttaa 2340 attatgggga ttttggagta tgtacatgat aaattataat acgtatatgg ttgaagttat 2400 tttatttttt actaatgaat tattttaata ttccttattg aataaatgct gtaacttgtt 2460 tgctatggaa cttattctta aagttctagt taaaaataat ttttccacat gcatgaaaat 2520 atgtattaat cagaggtggc ttaattacat tgaaattgct tttttgttgt tgttttttta 2580 ctgaaataac tcatgtttgt gtagaagaat gcctgtttac tcagagttta tattttcctt 2640 cagttatatt ttaaatcaaa aggtctgggt aatgtatact tttgattaat atatactttt 2700 tttaaaaaac aaaaaacaat gtaatggtta atagtagaaa tgtgccacac ttttcaagtt 2760 ttatataaca tatgaaattc agttaaaaga atgtgtgttt cataatgact tttaactggt 2820 aaaaatatta cttgcacgaa gtacttgatg tatggttatc ctgaaatttc ggagtatttg 2880 gtgtgttctt tgtctaaaaa tagtctgttt tgtcagtcct tcagaatatt atttattctg 2940 aagattgtcc ctcttgcact tggcagttta ttttcgggga tacattgttg ggggagaggg 3000 gtttctgcca ctctttccag attgagtctg tgctgtttaa ggaggactac catcctgcaa 3060 ctctttttct aattggggca cagaggatgt cgctaaagaa aagttgaaga gccctttcag 3120 cactttctca tctgtggaga agatggaatc ttaaaataca tttggagttt tatctgtttt 3180 acaagtccat tgatggccta agttcctcct gttttctgct gtttgatctc taaggaactc 3240 ctgttgctaa atatgaagag tatggcacat tcatatagtc tctgtgaagc atggggggag 3300 ggaagacatt tctttttctt ataggcttta tgctcaaatg tcatagtctc ctttcaaaga 3360 attgtgttgc attttaaatg cacccagctt aagtagaaga cattgaagga tgcattaatt 3420 ttcaggaact attttgaatt atgaaaagat tcccaattga aaaaattatt caacaagtaa 3480 aagctaagaa atttcattga aatcataagg cagtttaagc ataaattgat aaaaatagct 3540 gtgtactact aattaataga aaatcattca accaagagaa gagtcaagtg aatatcgttt 3600 gtttatttgc tagtgagttt ctttgtaacg ttgattttat taaatgataa tatttggtta 3660 gtatgtccta tgttaataaa aatgaacaaa attaattttg ctatgttcag gtgtcttgat 3720 aaaataacaa tgctccagtg ttgttgctta catttagcac taaattttaa cacagggtca 3780 gtgagtccag gttttaactt cttcatgcct ggatgggata aaatgtaatt cattgttaaa 3840 ttaattcata tttgtattta ttaatcactg tgacaacatt saccatttgt tcttaccagg 3900 aagtggtcag attatcatct gagttacagt tagactggct aagtttggta ttagatcaag 3960 gggaatgtcc agtaaacaga gaggtaagca tgatggaaat aatgaagtgg ggtacacagg 4020 aaaaacctga ctagtgagga ggagcagctg agagataggg tcagtgaatg cggttcagcc 4080 tgctacctct cctgtcttca tagaaccatk gccttagaat tattgtatga cacgtttttt 4140 gttggttaag ctgtaaggtt ttgttctttg tgaacatggg tattttgagg ggagggtgga 4200 gggagtaggg aagtggtcct tttacaagaa ttttgatgca taagtgtcta ttgtagggtt 4260 tggatgatct agtaaagtgt tttagaaccc ctttttatcc catgcaccat tcagtaaaca 4320 taaaaatcac aattctgnta atgtcatttg gaacttcaaa ataaatatct tgtctaaaaa 4380 caaaaaaaaa aaaaaaaa 4398 <210> 8 <211> 361 <212> PRT
<213> Homo sapiens <400> B
Met Ala Met Asn Tyr Asn Ala Lys Asp Glu Val Asp Gly Gly Pro Pro Cys Ala Pro Gly Gly Thr Ala Lys Thr Arg Arg Pro Asp Asn Thr Ala Phe Lys Gln Gln Arg Leu Pro Ala Trp Gln Pro Ile Leu Thr Ala Gly Thr Val Leu Pro Ile Phe Phe Ile Ile Gly Leu Ile Phe Ile Pro Ile Gly Ile Gly Ile Phe Val Thr Ser Asn Asn Ile Arg Glu Ile Glu Ile Asp Tyr Thr Gly Thr Glu Pro Ser Ser Pro Cys Asn Lys Cys Leu Ser Pro Asp Val Thr Pro Cys Phe Cys Thr Ile Asn Phe Thr Leu Glu Lys Ser Phe Glu Gly Asn Val Phe Met Tyr Tyr Gly Leu Ser Asn Phe Tyr Gln Asn His Arg Arg Tyr Val Lys Ser Arg Asp Asp Ser Gln Leu Asn Gly Asp Ser Ser Ala Leu Leu Asn Pro Ser Lys Glu Cys Glu Pro Tyr Arg Arg Asn Glu Asp Lys Pro Ile Ala Pro Cys Gly Ala Ile Ala Asn Ser Met Phe Asn Asp Thr Leu Glu Leu Phe Leu Ile Gly Asn Asp Ser Tyr Pro Ile Pro Ile Ala Leu Lys Lys Lys Gly Ile Ala Trp Trp Thr Asp Lys Asn Val Lys Phe Arg Asn Pro Pro Gly Gly Asp Asn Leu Glu Glu Arg Phe Lys Gly Thr Thr Lys Pro Val Asn Trp Leu Lys Pro Val Tyr Met Leu Asp Ser Asp Pro Asp Asn Asn Gly Phe Ile Asn Glu Asp Phe Ile Val Trp Met Arg Thr Ala Ala Leu Pro Thr Phe Arg Lys Leu Tyr Arg Leu Ile Glu Arg Lys Ser Asp Leu His Pro Thr Leu Pro Ala Gly Arg Tyr Ser Leu Asn Val Thr Tyr Asn Tyr Pro Val His Tyr Phe Asp Gly Arg Lys Arg Met Ile Leu Ser Thr Ile Ser Trp Met Gly Gly Lys Asn Pro Phe Leu Gly Ile Ala Tyr Ile Ala Val Gly Ser Ile Ser Phe Leu Leu Gly Val Val Leu Leu Val Ile Asn His Lys Tyr Arg Asn Ser Ser Asn Thr Ala Asp Ile Thr Ile <210> 9 <211> 777 <212> DNA
<213> Homo Sapiens <400> 9 agaaattata gagtgagtgt attttccttg tgtatatatt aaacacaccc atacaaacat 60 tggtaaagtt gattacatca aagaatcttt agcttatctt ttgaagctac tggatattat 120 tggtctctct aggtttttat ataaatagtg aaatctgaat tactgaaaac catgttaatt 180 tttagaactc attttcctca gtagagacta gtgatgcatt agcttctggg aacaaacttg 240 tatcggttct taattaaatt atccaaaacg gaggcattta aacacttgga tttacaccag 300 tcttttgtgt ttgcttttta aaataaagtg ctcgtatttg tattctccat attttggagt 360 aattatctac atgatgttta tagttcctgt ggtttttcac ccaagaagca gaatctcatt 420 cagtacattt agttttntaa gagtcatgaa gctaaatcct tgggctatgt cagaggcaca 480 aagtctagaa tgtgtgtatt cacnatggtg tatgtncatt ttgtgccttg attcacttag 540 aagtgtctca gaaaacctgg acngttcgct tctacacaag aattttatat gtatttatga 600 agatgattct gtaccctagt atatcttttt gggcatggac taatttgtat ctgtttaact 660 catattctgc acgatctgta tatagtacat caaacttaga ggtgtgacct taaatttaac 720 tttttttaaa aactgggagg tcaataaaat ttaaactgct taaaaaaaaa aaaaaaa 777 <210> 10 <211> 36 <212> PRT
<213> Homo sapiens <400> 10 Met Lys Met Ile Leu Tyr Pro Ser Ile Ser Phe Trp Ala Trp Thr Asn Leu Tyr Leu Phe Asn Ser Tyr Ser Ala Arg Ser Val Tyr Ser Thr Ser Asn Leu Glu Val <210> 11 <211> 1378 <212> DNA
<213> Homo Sapiens <400> 11 ctccgctggc aacggcgccg ctccccgctc ctcctcccca gccatggcgt tcacgttcgc 60 ggccttctgc tacatgctgg cgctgctgct cactgccgcg ctcatcttct tcgccatttg 120 gcacattata gcatttgatg agctgaagac tgattacaag aatcctatag accagtgtaa 180 taccctgaat ccccttgtac tcccagagta cctcatccac gctttcttct gtgtcatgtt 240 tctttgtgca gcagagtggc ttacactggg tctcaatatg cccctcttgg catatcatat 300 ttggaggtat atgagtagac cagtgatgag tggcccagga ctctatgacc ctacaaccat 360 catgaatgca gatattctag catattgtca gaaggaagga tggtgcaaat tagcttttta 420 tcttctagca tttttttact acctatatgg catgatctat gttttggtga gctcttagaa 480 caacacacag aagaattggt ccagttaagt gcatgcaaaa agccaccaaa tgaagggatt 540 ctatccagca agatcctgtc caagagtagc ctgtggaatc tgatcagtta ctttaaaaaa 600 tgactcctta ttttttaaat gtttccacat ttttgcttgt ggaaagactg ttttcatatg 660 ttatactcag ataaagattt taaatggtat tacgtataaa ttaatataaa atgattacct 720 ctggtgttga caggtttgaa cttgcacttc ttaaggaaca gccataatcc tctgaatgat 780 gcattaatta ctgactgtcc tagtacattg gaagcttttg tttataggaa cttgtagggc 840 tcattttggt ttcattgaaa cagtatctaa ttataaatta gctgtagata tcaggtgctt 900 ctgatgaagt gaaaatgtat atctgactag tgggaaactt catgggtttc ctcatctgtc 960 atgtcgatga ttatatatgg atacatttac aaaaataaaa agcgggaatt ttcccttcgc 1020 ttgaatatta tccctgtata ttgcatgaat gagagatttc ccatatttcc atcagagtaa 1080 taaatatact tgctttaatt cttaagcata agtaaacatg atataaaaat atatgctgaa 1140 ttacttgtga agaatgcatt taaagctatt ttaaatgtgt ttttatttgt aagacattac 1200 ttattaagaa attggttatt atgcttactg ttctaatctg gtggtaaagg tattcttaag 1260 aatttgcagg tactacagat tttcaaaact gaatgagaga aaattgtata accatcctgc 1320 tgttccttta gtgcaataca ataaaactct gaaattaaga ctcaaaaaaa aaaaaaaa 1378 <210> 12 <211> 144 <212> PRT
<213> Homo Sapiens <400> 12 Met Ala Phe Thr Phe Ala Ala Phe Cys Tyr Met Leu Ala Leu Leu Leu Thr Ala Ala Leu Ile Phe Phe Ala Ile Trp His Ile Ile Ala Phe Asp Glu Leu Lys Thr Asp Tyr Lys Asn Pro Ile Asp Gln Cys Asn Thr Leu Asn Pro Leu Val Leu Pro Glu Tyr Leu Ile His Ala Phe Phe Cys Val Met Phe Leu Cys Ala Ala Glu Trp Leu Thr Leu Gly Leu Asn Met Pro Leu Leu Ala Tyr His Ile Trp Arg Tyr Met Ser Arg Pro Val Met Ser Gly Pro Gly Leu Tyr Asp Pro Thr Thr Ile Met Asn Ala Asp Ile Leu Ala Tyr Cys Gln Lys Glu Gly Trp Cys Lys Leu Ala Phe Tyr Leu Leu Ala Phe Phe Tyr Tyr Leu Tyr Gly Met Ile Tyr Val Leu Val Ser Ser <210> 13 <211> 1450 <212> DNA
<213> Homo Sapiens <400> 13 1~

cgaaaatgtc aagagaaagg ccaaagattt gtactttttc acttacaaag cactcctttt 60 tcccttaaac ttctttctgt caaattagat ttaatgagag agtactattt ttaaggagct 120 atctgtttat gtagaatgat tttgttaaga gtaatgtaaa ctattattga gtagaggcct 180 aaagaggact gtgcattttt gctatttaaa ggaatcacaa atgatcatac ttaagtgagc 240 aaaaatgaca agttttacta gctaagtaga gaaataaatc tcaaatgcag cgctacaatt 300 ttcattatct taagtacatt gtacatttct acagaacctg tgattattct cgcatgataa 360 ggatggtact tgcatatggt gaattactac tgttgacagt ttccgcagaa atcctatttc 420 agtggaccaa cattgtggca tggcagcaaa tgccaacatt ttgtggaata gcagcaaatc 480 tacaagagac cctggttggt ttttcgtttt gttttctttg ttttttcccc cttctcctga 540 atcagcaggg atggaaggag ggtagggaag ttatgaatta ctccttccag tagtagctct 600 gaagtgtcac atttaatatc agtttttttt aaacatgatt ctagttaaat gtagaagaga 660 gaagaaagag gaagtgttca cttttttaat acactgattt agaaatttga tgtcttatat 720 cagtagttct gaggtattga tagcttgctt tatttctgcc tttacgttga cagtgttgaa 780 gcagggtgaa taactagggc atatattttt tttttttttg taagctgttt catgatgttt B40 tctttggaat ttccggataa gttcaggaaa acattctgca tgttgtatct agtctgatgt 900 acttatccat ctcattacaa acaaaaacac acagactgca tttgtagctc tgtaatcctt 960 gaatacggaa gtaaattttc ttctttcctg actttgacat tgtagctata ctgtttccat 1020 ttttgttttt acaaatcctt tgggtctaat tctgtgagcc tacctatagc actggattaa 1080 aatgtctgca tcatttcttt agttatccag ttaactttaa aactgttgta aaagtgtaaa 1140 ccagcccatg acaggttttt gwacatgtta aagaacttca ttgttcagtt ttcatgatta 1200 ttgtgtaagg aagactgatg tagatgttct gtgctgtcct ggaccatgtt aattacactt 1260 acgacgtatt ttagttccac atcacaatga tttgtcccca gtgacccttt tatcctttct 1320 aggcacattt cttgttgttg ttgttgttgc agttcccctt tgcattgtat tgctttgaca 1380 actgtaattt gaatcagatc tgaaagaggt ccagaataaa atatattttg atattaaaaa 1440 aaaaaaaaaa 1450 <210> 14 <211> 102 <212> PRT
<213> Homo sapiens <400> 14 Met Gln Arg Tyr Asn Phe His Tyr Leu Lys Tyr Ile Val His Phe Tyr Arg Thr Cys Asp Tyr Ser Arg Met Ile Arg Met Val Leu Ala Tyr Gly Glu Leu Leu Leu Leu Thr Val Ser Ala Glu Ile Leu Phe Gln Trp Thr Asn Ile Val Ala Trp Gln Gln Met Pro Thr Phe Cys Gly Ile Ala Ala Asn Leu Gln Glu Thr Leu Val Gly Phe Ser Phe Cys Phe Leu Cys Phe Phe Pro Leu Leu Leu Asn Gln Gln Gly Trp Lys Glu Gly Arg Glu Val Met Asn Tyr Ser Phe Gln <210> 15 <211> 3546 <212> DNA
<213> Homo sapiens <400> 15 ll gtggaattgg acactggtgc cccaagcgag gagttgagca gtgctggaga agtaacgaaa 60 cagacagtct tacmgaagga agaggagagg agtcagccaa ctaaaacccc ttcatcttct 120 caagagcccc ctgatgaagg aacctcaggg acagatgtga acaaaggatc mtcaaagaat 180 gctttgtcct cmatggatcc tgaagtgagg cttagtagcc ccccagggaa gccagaagat 240 tcatccagtg ttgatggtca gtcagtgggg actccagttg ggccagaaac tggaggagag 300 aagaatgggc cmgaagaaga ggaagaagag gactttgatg acctcaccca agatgaggaa 360 gatgaaatgt catcagcttc tgaggaatct gtgctttctg tcccagaact ccaggaaacm 420 atggagaaac tgacttggct ggcatctgaa aggcgcatga gtcaggaggg tgagtctgaa 480 gaagagaatt ctcaggagga gaactctgag ccagaagaag aggaggaaga agaagcagaa 540 ggaatggaaa gcctgcagaa agaggatgaa atgacggatg aagcagttgg agactctgct 600 gagaagcctc ctacttttgc ttcacctgag actgctccag aagtggagac cagcagaact 660 ccaccaggag agagcatcaa agctgctgga aaaggccgga acaatcatcg agctcgcaac 720 aagcggggaa gtcgggctcg ggccagcaag gacacctcca agctgctgtt gctgtatg~t 780 gaggacattc tcgagcgaga tccactcagg gagcagaagg acttggcctt tgcccaagct 840 tatctgacca gggtgcgaga agccctacaa catatccctg gcaagtatga agacttcctt 900 caagtcatct atgaatttga gtcaagtacc cagagacgga cggctgtaga tctctacaaa 960 agcctgcaaa ttctgctcca agactggcct cagctgttga aagactttgc tgctttcctg 1020 ttacctgagc aagctctggc ctgtggatta tttgaggagc agcaggcttt tgagaagagc 1080 cgcaagttcc ttcggcagct ggagatttgc tttgcagaga acccctcaca ccaccagaag 1140 attatcaagg tcctccaagg ctgtgcagac tgccttcccc aggagatcac cgagctcaag 1200 acacagatgt ggcagctcct caagggccac gaccacctgc aggatgagtt ttctatcttc 1260 tttgaccact tgcgcccagc agctagccgg atgggtgact ttgaagagat caattggact 1320 gaggaaaagg agtatgagtt tgatggcttt gaagaagtgg ccctgcctga tgtggaagaa 1380 gaggaggagc ctcccaagat acccacagcc tcaaagaaca agaggaaaaa agagatcggg 1440 gtccaaaatc atgataagga gactgaatgg ccagatgggg ccaaggactg tgcctgctcc 1500 tgccatgaag gaggtccaga ttccaagctg aagaagagca aaaggcggag ctgtagccac 1560 tgtagcagca aggtctgtga cagcaaatcc tacaagagca aggagcccca tgagttggtg 1620 ggcagcagcc cccaccgaga ggctagtcct atgcctggtg ctaaggaagc tgggcagggc 1680 aaggatatga tggaagagga agccccagag gagcgggaga gcactgaggc cacccagagc 1740 aggactgtca ggaccaccag aaagggagag atgcctgttt caggattggc agtggggagc 1800 actttgccat cccctcgaga agtgactgtt acagaacggc tcctcctgga tggcccacca 1860 cctcattcac cagagactcc tcaatttccc cccacaactg gagctgtact gtacactgtt 1920 aagagaaacc aggttgggcc tgaggttcgc tcctgcccca aggcatcccc cagacttcag 1980 aaagagaggg agggccaaaa ggcagtgagt gagtcagagg ctttgatgct ggtctgggat 2040 gcatcagaaa ctgagaaatt gcctggtacc gtgggaaccc cctgcttcct tcctgagtcc 2100 tgtttcctca aagaccagag atgcagggag aagacatgtg tccgggaaac cagacactca 2160 agagagatgg ctgccctcaa gcagagctcg ggtgaagaca agagacagga cgtgccctgt 2220 ccatgaatct ccatcaggaa ttgacacctc agagacttct cccaaagccc ctagaggggg 2280 tttggctaaa gacagtggaa cacaggccaa gggtccagag ggggagcagc agccaaaggc 2340 cgcagaagct acggtgtgtg ccaacaacag caaggtcagc tccactgggg aaaaggttgt 2400 cctgtggaca agggaagctg accgtgtgat cctcaccatg tgccaggagc aaggggcaca 2460 gccacagacc ttcaacatca tctcccagca gctgggaaat aagacccctg ctgaggtttc 2520 ccaccgtttt cgagaactca tgcagctctt ccacactgcc tgtgaagcca gctctgagga 2580 tgaggatgat gcaaccagta ccagcaatgc agaccagctg tctgaccatg gggaccttct 2640 gtctgaagag gagctggatg aatgagactc tgggaatcat ctacacagga ccaaacccaa 2700 caggcgccct ggcaccgggg agggggtagt tgtactctgc ttgtacagtc cttgagccca 2760 gtttacagat ctggagagca ggaggccagg acaaggacaa aggctggagg atggagtagg 2820 acccaggggc tctgccatcc taggcatcat tcaaggtctt ttatgaagac tttacagatg 2880 tcctctgtaa atagcatcga gagtggagtt cagctccttt ctctactttt ttttggtctg 2940 atggcacata tttattgttc tgtggtctaa tcacagtgtt tctaaatgta aaaagtgcat 3000 atgttggtgt agctagtccc gcgacattga gctcctctgc atgaagacac tgggctcctg 3060 catccagctg tttttattgc aaactagctc ctttctccca cactgggaac tttagtccac 3120 gaggctgtca ccaccctggt agcactgggc caggctttgt agctcctgca gcagctctgc 3180 tacgtcatcg tgctccactc cagcatccat gaagctggcc cagcgccgca agtcgagttt 3240 ggtgaggtct ctggccaagg cttccagggt ctggtgcagg gacaaagagg aacacagtgc 3300 cccaaacact gggatgctct ccactgctgt ggagggagag gaaacagaga cctgtagatg 3360 gatgattatt ctgccctggg actcgccaaa ctgataagga agtccaacct tagtagactt 3420 gattgtaaac tcaacaaatt tggtgtattg tccccttagt acaccagtac tccagaggaa 3480 gaatgctttt cttgggagcc atagggtgaa taaaggaatg tttaactgtg aaaaaaaaaa 3540 aaaaaa 3546

12 <210> 16 <211> 677 <212> PRT
<213> Homo sapiens <400> 16 Met Asp Pro Glu Val Arg Leu Ser Ser Pro Pro Gly Lys Pro Glu Asp Ser Ser Ser Val Asp Gly Gln Ser Val Gly Thr Pro Val Gly Pro Glu Thr Gly Gly Glu Lys Asn Gly Pro Glu Glu Glu Glu Glu Glu Asp Phe Asp Asp Leu Thr Gln Asp Glu Glu Asp Glu Met Ser Ser Ala Ser Glu Glu Ser Val Leu Ser Val Pro Glu Leu Gln Glu Thr Met Glu Lys Leu Thr Trp Leu Ala Ser Glu Arg Arg Met Ser Gln Glu Gly Glu Ser Glu Glu Glu Asn Ser Gln Glu Glu Asn Ser Glu Pro Glu Glu Glu Glu Glu Glu Glu Ala Glu Gly Met Glu Ser Leu Gln Lys Glu Asp Glu Met Thr Asp Glu Ala Val Gly Asp Ser Ala Glu Lys Pro Pro Thr Phe Ala Ser Pro Glu Thr Ala Pro Glu Val Glu Thr Ser Arg Thr Pro Pro Gly Glu Ser Ile Lys Ala Ala Gly Lys Gly Arg Asn Asn His Arg Ala Arg Asn Lys Arg Gly Ser Arg Ala Arg Ala Ser Lys Asp Thr Ser Lys Leu Leu Leu Leu Tyr Asp Glu Asp Ile Leu Glu Arg Asp Pro Leu Arg Glu Gln Lys Asp Leu Ala Phe Ala Gln Ala Tyr Leu Thr Arg Val Arg Glu Ala Leu Gln His Ile Pro Gly Lys Tyr Glu Asp Phe Leu Gln Val Ile Tyr Glu Phe Glu Ser Ser Thr Gln Arg Arg Thr Ala Val Asp Leu Tyr Lys Ser Leu Gln Ile Leu Leu Gln Asp Trp Pro Gln Leu Leu Lys Asp Phe Ala Ala Phe Leu Leu Pro Glu Gln Ala Leu Ala Cys Gly Leu Phe Glu

13 Glu Gln Gln Ala Phe Glu Lys Ser Arg Lys Phe Leu Arg Gln Leu Glu Ile Cys Phe Ala Glu Asn Pro Ser His His Gln Lys Ile Ile Lys Val Leu Gln Gly Cys Ala Asp Cys Leu Pro Gln Glu Ile Thr Glu Leu Lys Thr Gln Met Trp Gln Leu Leu Lys Gly His Asp His Leu Gln Asp Glu Phe Ser Ile Phe Phe Asp His Leu Arg Pro Ala Ala Ser Arg Met Gly Asp Phe Glu Glu Ile Asn Trp Thr Glu Glu Lys Glu Tyr Glu Phe Asp Gly Phe Glu Glu Val Ala Leu Pro Asp Val Glu Glu Glu Glu Glu Pro Pro Lys Ile Pro Thr Ala Ser Lys Asn Lys Arg Lys Lys Glu Ile Gly Val Gln Asn His Asp Lys Glu Thr Glu Trp Pro Asp Gly Ala Lys Asp Cys Ala Cys Ser Cys His Glu Gly Gly Pro Asp Ser Lys Leu Lys Lys Ser Lys Arg Arg Ser Cys Ser His Cys Ser Ser Lys Val Cys Asp Ser Lys Ser Tyr Lys Ser Lys Glu Pro His Glu Leu Val Gly Ser Ser Pro His Arg Glu Ala Ser Pro Met Pro Gly Ala Lys Glu Ala Gly Gln Gly Lys Asp Met Met Glu Glu Glu Ala Pro Glu Glu Arg Glu Ser Thr Glu Ala Thr Gln Ser Arg Thr Val Arg Thr Thr Arg Lys Gly Glu Met Pro Val Ser Gly Leu Ala Val Gly Ser Thr Leu Pro Ser Pro Arg Glu Val Thr Val Thr Glu Arg Leu Leu Leu Asp Gly Pro Pro Pro His Ser Pro 545 550 ~ 555 560 Glu Thr Pro Gln Phe Pro Pro Thr Thr Gly Ala Val Leu Tyr Thr Val Lys Arg Asn Gln Val Gly Pro Glu Val Arg Ser Cys Pro Lys Ala Ser Pro Arg Leu Gln Lys Glu Arg Glu Gly Gln Lys Ala Val Ser Glu Ser

14 Glu Ala Leu Met Leu Val Trp Asp Ala Ser Glu Thr Glu Lys Leu Pro Gly Thr Val Gly Thr Pro Cys Phe Leu Pro Glu Ser Cys Phe Leu Lys Asp Gln Arg Cys Arg Glu Lys Thr Cys Val Arg Glu Thr Arg His Ser Arg Glu Met Ala Ala Leu Lys Gln Ser Ser Gly Glu Asp Lys Arg Gln Asp Val Pro Cys Pro <210> 17 <211> 3052 <212> DNA
<213> Homo sapiens <900> 17 atgatgcgtc cctgcctcgg ccgctgcagt cgccgccgcc gccgccgcag gccgggagga 60 gccgcagcgc cgggcgaccc cgcccgggcc tcggatccga tcacatagga cagtatgcac 120 cttaagatcc tgaagaaacg gcacaaaatg ttcaagtgat gtttagaaat aacttgtgag 180 ggtgcgtcag ggaaatcatg cagccatcag gacacaggct ccgggacgtc gagcatcatc 240 ctctcctggc tgaaaatgac aactatgact cttcatcgtc ctcctcctcc gaggctgacg 300 tggctgaccg ggtctggttc atccgtgacg gctgcggcat gatctgtgct gtcatgacgt 360 ggcttctggt cgcctatgca gacttcgtgg tgactttcgt catgctgctg ccttccaaag 420 acttctggta ctctgtggtc aacggggtca tctttaactg cttggccgtg cttgccctgt 480 catcccacct gagaaccatg ctcaccgacc ctggggcagt acccaaagga aacgctacga 540 aagaatacat ggagagcttg cagctgaagc ccggggaagt catctacaag tgccccaagt 600 gctgctgtat taaacccgag cgcgcccacc actgcagtat ttgcaaaaga tgtattcgga 660 aaatggatca tcactgcccg tgggtgaaca attgtgtagg agaaaagaat caaagatttt 720 ttgtgctctt cactatgtat atagctctgt cttcagtcca tgctctgatc ctttgtggat 780 ttcagttcat ctcctgtgtc cgagggcagt ggactgaatg cagtgatttt tcacctccga 840 taactgtaat cctgttgatc ttcctgtgcc ttgagggtct tctgtttttc actttcactg 900 cagttatgtt tggcacccaa atccactcca tatgcaacga cgagacggag atcgagcgat 960 tgaaaagtga gaagcccaca tgggagcgga ggctgcgatg ggaagggatg aagtccgtct 1020 ttggggggcc cccctcactc ctctggatga atccctttgt gggcttccga tttaggcgac 1080 tgcccacgag acccagaaaa ggtggcccgg agttctcagt gtgaggcgtg gctcatcaga 1140 ctgaaacttg ctcacagact tccagttatt tatttggggt ctgsaggata tcaacagctc 1200 atctgtgacc aacagggcaa ctggaaccta cacaaaccaa ttgcttgcag caagcagagt 1260 tttatatatt tatagtcaca gatggcagag gaagaggctc tcagtcccca cctgtacaac 1320 aacggaaagg tgtgtggcca cacgaagaag ccaaacgccg tggcctcctg cagagctggg 1380 gcttctgtgg agaatacttc gggttattac atgggttatt caaatcctgg gtcctgagct 1440 gctgtttcca atcatgaaga aaaacagtga atccagtgaa cagggattct ccaagcagtc 1500 atttcagggg gctcctgctg accccgccac tcagcagtgc actccccgga tcacagcagg 1560 gcgtttacat agaaagacgt tttggtctcg attagctccg atgctttgca ctgaagttgc 1620 aaaagatctg tgcactgaac agtgaaggtg gcttccggca cactccccgc tgccccggaa 1680 gagacatcct ttgaccctct cagcaagtct gtgtgtgtgc gtgtctgtgc gtgtgcgcgc 1740 gtgtgtgcat gtgtgtcaaa attgccagtg ttgtttaggc aatgtaacat ttaccggctg 1800 tgtacagcaa acaagctatt ttttagaaac cgacgtttca gggaagaggg gagagagccg 1860 cggggtcctg cccgtggtta ctatgaatgt attgctgttg gaggacatct cgatccaaag 1920 aacagccgtt cctgtgcggc ccttcgttgc cctcctgctt tcatttttta aagaaatctt 1980 gagtgcttga gggccttgga actgattttt tttttttgtt ccagccaaat tagcagtgta 2040 taaatggcac ctaggtaaga gcagagctgc ggctcggtga cttgatactt ggggcagccc 2100 gatgctctgt gtggggcagg ggaggcatcc ttactggaga ggcagggccc agccattggg 2160 cacctctggg aaggggaggg gaccatgagg cagccagccc ctggcagggg cgactgtgcc 2220 accgcaggca gcgctccagt cgggaatggc caggatggcg ccctcttgtt ggagtttttg 2280 gttagctttt acgttttctt ctccacccac ggcacaggtg ataaaatagg atccttggtg 2340 cggagcttaa aattatgcca gaaagccaac agctcccctc gtggggcctt gccttaaact 2400 tgcttggttt gtacattttt tgccggacgc atcaagaagc aatctgtgac aaagtctgag 2460 ggtcttcctt tatgcttgcc ctccacatta agagaagttg ggtctccctc ctgggaattg 2520 tttggccttt ctgttcatct gtgaactgtt ttttgttttt aattactctg taccccatcc 2580 gaatcagggc ttctaccact gctgatgcaa aaccacaaag ggacctacct gagccaccgt 2640 cctagccaag cgagcaaacc tgcagggggt ttggaagtgg acttggtcac cgcagaagcg 2700 tgtgcgccgt tgggggaaga gctgcgtcac agccagaggg acaaagtgtg ggtgatcctg 2760 gagacgccag tttccgagat tgttctgcat attcatttgc acattgttgt ctgggttgga 2820 catgcgtgtg ggcttcagtg tgaggctttt aatatgtata tcctgttatc aataaaacaa 2880 ttatccaagt ggttgaatcc tgtgagactt ggcaagtgtg tgcaaatcaa gtatacttga 2940 cttttcaacc tcttctttca atgtaacttt tatatgaaat aaagtaatca attaacagtt 3000 ctcaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa as 3052 <210> 18 <211> 308 <212> PRT
<213> Homo sapiens <400> 18 Met Gln Pro Ser Gly His Arg Leu Arg Asp Val Glu His His Pro Leu Leu Ala Glu Asn Asp Asn Tyr Asp Ser Ser Ser Ser Ser Ser Ser Glu Ala Asp Val Ala Asp Arg Val Trp Phe Ile Arg Asp Gly Cys Gly Met Ile Cys Ala Val Met Thr Trp Leu Leu Val Ala Tyr Ala Asp Phe Val Val Thr Phe Val Met Leu Leu Pro Ser Lys Asp Phe Trp Tyr Ser Val Val Asn Gly Val Ile Phe Asn Cys Leu Ala Val Leu Ala Leu Ser Ser His Leu Arg Thr Met Leu Thr Asp Pro Gly Ala Val Pro Lys Gly Asn Ala Thr Lys Glu Tyr Met Glu Ser Leu Gln Leu Lys Pro Gly Glu Val Ile Tyr Lys Cys Pro Lys Cys Cys Cys Ile Lys Pro Glu Arg Ala His His Cys Ser Ile Cys Lys Arg Cys Ile Arg Lys Met Asp His His Cys Pro Trp Val Asn Asn Cys Val Gly Glu Lys Asn Gln Arg Phe Phe Val Leu Phe Thr Met Tyr Ile Ala Leu Ser Ser Val His Ala Leu Ile Leu Cys Gly Phe Gln Phe Ile Ser Cys Val Arg Gly Gln Trp Thr Glu Cys Ser Asp Phe Ser Pro Pro Ile Thr Val Ile Leu Leu Ile Phe Leu Cys Leu Glu Gly Leu Leu Phe Phe Thr Phe Thr Ala Val Met Phe Gly Thr Gln Ile His Ser Ile Cys Asn Asp Glu Thr Glu Ile Glu Arg Leu Lys Ser Glu Lys Pro Thr Trp Glu Arg Arg Leu Arg Trp Glu Gly Met Lys Ser Val Phe Gly Gly Pro Pro Ser Leu Leu Trp Met Asn Pro Phe Val Gly Phe Arg Phe Arg Arg Leu Pro Thr Arg Pro Arg Lys Gly Gly Pro Glu Phe Ser Val <210> 19 <211> 3123 <212> DNA
<213> Homo sapiens <400> 19 ggatgctgga gcagaacaat ggatttctct ttctctttca tgcaagggat catgggaaac 60 acaattcagc aaccacctca actcattgac tccgccaaca tccgtcagga ggatgccttt 120 gataacaaca gtgacattgc tgaagatggt ggccagacac catatgaagc tactttgcag 180 caaggctttc agtacccagc tacaacagaa gatcttcctc cactcacaaa tgggtatcca 240 tcatcaatca gtgtgtatga aactcaaacc aaataccagt catataatca gtatcctaat 300 gggtcagcca atggctttgg tgcagttaga aactttagcc ccactgacta ttatcattca 360 gaaattccaa acacaagacc acatgaaatt ctggaaaaac cttcccctcc acagccacca 420 cctcctcctt cggtaccaca aactgtgatt ccaaagaaga ctggctcacc tgaaattaaa 480 ctaaaaataa ccaaaactat ccagaatggc agggaattgt ttgagtcttc cctttgtgga 540 gaccttttaa atgaagtaca ggcaagtgag cacacgaaat caaagcatga aagcagaaaa 600 gaaaagagga aaaaaagcaa caagcatgac tcatcaagat ctgaagagcg caagtcacac 660 aaaatcccca aattagaacc agaggaacaa aatagaccaa atgagagggt tgacactgta 720 tcagaaaaac caagggaaga accagtacta aaagaggaag ccccagttca gccaatacta 780 tcttctgttc caacaacgga agtgtccact ggtgttaagt ttcaggttgg cgatcttgtg 840 tggtccaagg tgggaaccta tccttggtgg ccttgtatgg tttcaagtga tccccagctt 900 gaggttcata ctaaaattaa cacaagaggt gcccgagaat atcatgtcca gttttttagc 960 aaccagccag agagggcgtg ggttcatgaa aaacgggtac gagagtataa aggtcataaa 1020 cagtatgaag aattactggc tgaggcaacc aaacaagcca gcaatcactc tgagaaacaa 1080 aagattcgga aaccccgacc tcagagagaa cgtgctcagt gggatattgg cattgcccat 1140 gcagagaaag cattgaaaat gactcgagaa gaaagaatag aacagtatac ttttatttac 1200 attgataaac agcctgaaga ggctttatcc caagcaaaaa agagtgttgc ctccaaaacc 1260 gaagttaaaa aaacccgacg accaagatct gtgctgaata ctcagccaga acagaccaat 1320 gcaggggagg tggcctcctc actctcaagt actgaaattc ggagacatag ccagaggcgg 1380 cacacaagtg cggaagagga agagccaccg cctgttaaaa tagcctggaa aactgcggca 1440 gcaaggaaat ccttaccagc ttccattacg atgcacaaag ggagcctgga tttgcagaag 1500 tgtaacatgt ctccagttgt gaaaattgaa caagtgtttg ctcttcagaa tgctacaggg 1560 gatgggaaat ttatcgatca atttgtttat tcaacaaagg gaattggtaa caaaacagaa 1620 ataagtgtca gggggcaaga caggcttata atttctacac caaaccagag aaatgaaaag 1680 ccaacgcaga gtgtatcatc tcctgaagca acatctggtt ctacaggctc agtagaaaag 1740 aagcaacaga gaagatcaat tagaactcgt tctgaatcag agaaatccac tgaggttgtg 1800 ccaaagaaga agatcaaaaa ggagcaggtt gaaacagttc ctcaggctac agtgaagact 1860 ggattacaga aagggtcggc ggaccgggga gtgcagggct ctgtcagatt cagtgacagc 1920 tccgtctccg cagcgattga ggaaactgtg gactgagatt cctgtacaat ttcatcccag 1980 aaactccaga cttgtagtct ccatgcaaga tttctttgtc ggcggcttga taaacagttt 2040 ctttgttttc gattttgatt tcgccaatca tcattattgg cattttcctg cctggtttct 2100 tcttcaagac tctgaacaat tgctttaaca gtcaaatgat tttttttttc ggtttgagct 2160 ggatgggtac agcttaaatc atgggtccag cctaaaaacc accatttaac ttacactgat 2220 caatttcaac atggactgtt tttggttttt tgtttttaaa taaagcatca ttaatgcaca 2280 tctgcagggg tttgccaaac agcccaaact gtatacatta caatcattaa aagttcttat 2340 tttttttaat attagtgccg ttatcatgga gaacagcatg acagctgtct ttggcagtct 2400 gtcatttttc tagcattttc agaaactcat cggaaatggc ggtacctgtg tttcccttcg 2460 aaagcctctc agtacagcac tcctgttcct ctgttaaaac tccttgttaa tccagtgatc 2520~
ttttaggcca aggaaatatt ttgtgatggt gttctgggtc catacaccag caatgaagga 2580 gatagatttg tgtacttgtg ttttttaatc agcattaaca tgggcaggca ccctcattta 2640 tagatgtcag gaaacattca gtgaaaaact tgtagaatgg gatgtgataa cgaggttcca 2700 gtaatctgag cagtctaacg aggcccacct cctccaccac agaacgtggc tatgttccaa 2760 gtgctactct cactcagcct gttgcggatc ttcatggcct caggagactt gtttctccat 2820 gggctcttct ggactgcaca cttccaccat agcttgctgg gttgatctag atgtctgttt 2880 gttgtatgga aattttgggg gaaaaaatcc aaaacacaaa ctgtgggttg aaatattaac 2940 cgtctccttg gttccttggt attcaccgtg cctgatctgc acatttcatc gtggctgttt 3000 ctgtatagcc tatactgcat tagcccaaga gattgttgct ttgtaacttt ttgcactatt 3060 gttttggctg gatttgtatt acacacagtt ttaaaaaaaa caataaaaaa aaaaaaaaaa 3120 aaa <210> 20 <211> 645 <212> PRT
<213> Homo sapiens <400> 20 Met Asp Phe Ser Phe Ser Phe Met Gln Gly Ile Met Gly Asn Thr Ile Gln Gln Pro Pro Gln Leu Ile Asp Ser Ala Asn Ile Arg Gln Glu Asp Ala Phe Asp Asn Asn Ser Asp Ile Ala Glu Asp Gly Gly Gln Thr Pro Tyr Glu Ala Thr Leu Gln Gln Gly Phe Gln Tyr Pro Ala Thr Thr Glu Asp Leu Pro Pro Leu Thr Asn Gly Tyr Pro Ser Ser Ile Ser Val Tyr Glu Thr Gln Thr Lys Tyr Gln Ser Tyr Asn Gln Tyr Pro Asn Gly Ser Ala Asn Gly Phe Gly Ala Val Arg Asn Phe Ser Pro Thr Asp Tyr Tyr His Ser Glu Ile Pro Asn Thr Arg Pro His Glu Ile Leu Glu Lys Pro Ser Pro Pro Gln Pro Pro Pro Pro Pro Ser Val Pro Gln Thr Val Ile Pro Lys Lys Thr Gly Ser Pro Glu Ile Lys Leu Lys Ile Thr Lys Thr Ile Gln Asn Gly Arg Glu Leu Phe Glu Ser Ser Leu Cys Gly Asp Leu Leu Asn Glu Val Gln Ala Ser Glu His Thr Lys Ser Lys His Glu Ser Arg Lys Glu Lys Arg Lys Lys Ser Asn Lys His Asp Ser Ser Arg Ser Glu Glu Arg Lys Ser His Lys Ile Pro Lys Leu Glu Pro Glu Glu Gln Asn Arg Pro Asn Glu Arg Val Asp Thr Val Ser Glu Lys Pro Arg Glu Glu Pro Val Leu Lys Glu Glu Ala Pro Val Gln Pro Ile Leu Ser Ser Val Pro Thr Thr Glu Val Ser Thr Gly Val Lys Phe Gln Val Gly Asp Leu Val Trp Ser Lys Val Gly Thr Tyr Pro Trp Trp Pro Cys Met Val Ser Ser Asp Pro Gln Leu Glu Val His Thr Lys Ile Asn Thr Arg Gly Ala Arg Glu Tyr His Val Gln Phe Phe Ser Asn Gln Pro Glu Arg Ala Trp Val His Glu Lys Arg Val Arg Glu Tyr Lys Gly His Lys Gln Tyr Glu Glu Leu Leu Ala Glu Ala Thr Lys Gln Ala Ser Asn His Ser Glu Lys Gln Lys Ile Arg Lys Pro Arg Pro Gln Arg Glu Arg Ala Gln Trp Asp Ile Gly Ile Ala His Ala Glu Lys Ala Leu Lys Met Thr Arg Glu Glu Arg Ile Glu Gln Tyr Thr Phe Ile Tyr Ile Asp Lys Gln Pro Glu Glu Ala Leu Ser Gln Ala Lys Lys Ser Val Ala Ser Lys Thr Glu Val Lys Lys Thr Arg Arg Pro Arg Ser Val Leu Asn Thr Gln Pro Glu Gln Thr Asn Ala Gly Glu Val Ala Ser Ser Leu Ser Ser Thr Glu Ile Arg Arg His Ser Gln Arg Arg His Thr Ser Ala Glu Glu Glu Glu Pro Pro Pro Val Lys Ile Ala Trp Lys Thr Ala Ala Ala Arg Lys Ser Leu Pro Ala Ser Ile Thr Met His Lys Gly Ser Leu Asp Leu Gln Lys Cys Asn Met Ser Pro Val Val Lys Ile Glu Gln Val Phe Ala Leu Gln Asn Ala Thr Gly Asp Gly Lys Phe Ile Asp Gln Phe Val Tyr Ser Thr Lys Gly Ile Gly Asn Lys Thr Glu Ile Ser Val Arg Gly Gln Asp Arg Leu Ile Ile Ser Thr Pro Asn Gln Arg Asn Glu Lys Pro Thr Gln Ser Val Ser Ser Pro Glu Ala Thr Ser Gly Ser Thr Gly Ser Val Glu Lys Lys Gln Gln Arg Arg Ser Ile Arg Thr Arg Ser Glu Ser Glu Lys Ser Thr Glu Val Val Pro Lys Lys Lys Ile Lys Lys Glu Gln Val Glu Thr Val Pro Gln Ala Thr Val Lys Thr Gly Leu Gln Lys Gly Ser Ala Asp Arg Gly Val Gln Gly Ser Val Arg Phe Ser Asp Ser Ser Val Ser Ala Ala Ile Glu Glu Thr Val Asp <210> 21 <211> 29 <212> DNA
<213> Artificial Sequence <220>
<223> oligonucleotide <220>
<221> misc_feature <222> (2) <223> biotinylated phosphoaramidite residue <400> 21 gntggcattc caggtgtggt ggtgcacag 2g <210> 22 <211> 29 <212> DNA
<213> Artificial Sequence <220>
<223> oligonucleotide <220>
<221> misc_feature <222> (2) <223> biotinylated phosphoaramidite residue <400> 22 cnaggctaat gaggcagcaa tggcttaaa 29 <210> 23 <211> 21 <212> DNA

<213> Artificial Sequence <220>

<223> oligonucleotide <400> 23 tcttcactgt atctcctttg c 21 <210> 24 <211> 29 <212> DNA

<213> Artificial Sequence <220>

<223> oligonucleotide <220>

<221> misc_feature <222> (2) <223> biotinylated phosphoaramidite residue <400> 24 anaactccca gaaggaagga gatggatcc <210> 25 <211> 29 <212> DNA

<213> Artificial Sequence <220>

<223> oligonucleotide <220>

<221> misc_feature <222> (2) <223> biotinylated phosphoaramidite residue <400> 25 anattcttgt gtagaagcga actgtccag 29 <210> 26 <211> 29 <212> DNA

<213> Artificial Sequence <220>

<223> oligonucleotide <220>

<221> misc_feature <222> (2) <223> biotinylated phosphoaramidite residue <400> 26 anggcgaaga agatgagcgc ggcagtgag 29 <210> 27 <211> 29 <212> DNA
<213> Artificial Sequence <220>
<223> oligonucleotide <220>
<221> misc_feature <222> (2) <223> biotinylated phosphoaramidite residue <400> 27 gnaagaagtt taagggaaaa aggagtgct 29 <210> 28 <211> 29 <212> DNA
<213> Artificial Sequence <220>
<223> oligonucleotide <220>
<221> misc_feature <222> (2) <223> biotinylated phosphoaramidite residue <400> 28 cnctttttcc tcttgttctt tgaggctgt 29 <210> 29 <211> 29 <212> DNA
<213> Artificial Sequence <220>
<223> oligonucleotide <220>
<221> misc_feature <222> (2) <223> biotinylated phosphoaramidite residue <400> 29 anggatcaga gcatggactg aagacagag 29 <210> 30 <211> 29 <212> DNA
<213> Artificial Sequence <220>
<223> oligonucleotide <220>

<221> misc_feature <222> (2) <223> biotinylated phosphoaramidite residue <400> 30 anatagacca aatgagaggg ttgacactg 29 <210> 31 <211> 87 <212> PRT
<213> Homo Sapiens <400> 31 Met Glu Met Gly Ile Gln Val Gly Lys Arg Leu Leu Ile Gln Gly Leu Glu Leu Gly Glu Leu Leu Thr Lys Cys Gly Asn Glu Asp Phe Leu Lys Met Gly Ala Gly Trp Ile Gln Ala Ser Tyr Phe Asn Pro Gly Leu Thr Leu Ile Gln Ser Lys Tyr Phe Leu Ser Ser Ser Trp Gly Gly Val Leu Gly Gln Glu Lys Lys Gly Asp Lys Asn Thr Ile Pro Glu Lys Glu Glu Lys Tyr Leu His Phe Val Leu as <210> 32 <211> 82 <212> PRT
<213> Homo Sapiens <400> 32 Met Met Phe Ile Val Pro Val Val Phe His Pro Arg Ser Arg Ile Ser Phe Ser Thr Phe Ser Phe Ile Arg Val Met Lys Leu Asn Pro Trp Ala Met Ser Glu Ala Gln Ser Leu Glu Cys Val Tyr Ser Gln Trp Cys Met Tyr Ile Leu Cys Leu Asp Ser Leu Arg Ser Val Ser Glu Asn Leu Asp Ser Ser Leu Leu His Lys Asn Phe Ile Cys Ile Tyr Glu Asp Asp Ser Val Pro

Claims

What is claimed is:

1. An isolated polynucleotide selected from the group consisting of:
(a) a polynucleotide comprising the nucleotide sequence of SEQ ID
NO:1;
(b) a polynucleotide comprising the nucleotide sequence of SEQ ID
NO:1 from nucleotide 216 to nucleotide 2174;
(c) a polynucleotide comprising the nucleotide sequence of SEQ ID
NO:1 from nucleotide 339 to nucleotide 2174;
(d) a polynucleotide comprising the nucleotide sequence of the full-length protein coding sequence of done cc359 4 deposited under accession number ATCC 98715;
(e) a polynucleotide encoding the full-length protein encoded by the cDNA insert of clone cc359 4 deposited under accession number ATCC 98715;
(f) a polynucleotide comprising the nucleotide sequence of the mature protein coding sequence of clone cc359 4 deposited under accession number ATCC 98715;
(g) a polynucleotide encoding the mature protein encoded by the cDNA insert of clone cc359 4 deposited under accession number ATCC 98715;
(h) a polynucleotide encoding a protein comprising the amino acid sequence of SEQ ID NO:2;
(i) a polynucleotide encoding a protein comprising a fragment of the amino acid sequence of SEQ ID NO:2, the fragment comprising eight contiguous amino acids of SEQ ID NO:2;
(j) a polynucleotide that hybridizes under conditions at least as stringent as 4X SSC at 65 degrees C, or 4X SSC at 42 degrees C with 50%
formamide, to any one of the polynucleotides specified in (a)-(i); and (k) a polynucleotide that hybridizes under conditions at least as stringent as 4X SSC at 50 degrees C, or 6X SSC at 40 degrees C with 50%
formamide, to any one of the polynucleotides specified in (a)-(i), and that has a length that is at least 25% of the length of SEQ ID NO:1.

2. The polynucleotide of claim 1 wherein said polynucleotide is operably linked to at least one expression control sequence.

3. A host cell transformed with the polynucleotide of claim 2.

4. The host cell of claim 3, wherein said cell is a mammalian cell.

5. A process for producing a protein encoded by the polynucleotide of claim 2, which process comprises:
(a) growing a culture of the host cell of claim 3 in a suitable culture medium; and (b) purifying said protein from the culture.

6. A protein produced according to the process of claim 5.

7. An isolated polynucleotide encoding the protein of claim 6.

8. The polynucleotide of claim 7, wherein the polynucleotide comprises the cDNA insert of clone cc359_4 deposited under accession number ATCC 98715.

9. A protein comprising an amino acid sequence selected from the group consisting of:
(a) the amino acid sequence of SEQ ID NO:2;
(b) a fragment of the amino acid sequence of SEQ ID NO:2, the fragment comprising eight contiguous amino acids of SEQ ID NO:2; and (c) the amino acid sequence encoded by the cDNA insert of clone cc359_4 deposited under accession number ATCC 98715;
the protein being substantially free from other mammalian proteins.

10. The protein of claim 9, wherein said protein comprises the amino acid sequence of SEQ ID NO:2.

11. A composition comprising the protein of claim 9 and a pharmaceutically acceptable carrier.

12. An isolated polynucleotide selected from the group consisting of:

(a) a polynucleotide comprising the nucleotide sequence of SEQ ID
NO:3;
(b) a polynucleotide comprising the nucleotide sequence of SEQ ID
NO:3 from nucleotide 202 to nucleotide 414;
(c) a polynucleotide comprising the nucleotide sequence of SEQ ID
NO:3 from nucleotide 370 to nucleotide 414;
(d) a polynucleotide comprising the nucleotide sequence of the full-length protein coding sequence of clone ct547 2 deposited under accession number ATCC 98715;
(e) a polynucleotide encoding the full-length protein encoded by the cDNA insert of clone ct547_2 deposited under accession number ATCC 98715;
(f) a polynucleotide comprising the nucleotide sequence of the mature protein coding sequence of clone ct547_2 deposited under accession number ATCC 98715;
(g) a polynucleotide encoding the mature protein encoded by the cDNA insert of clone ct547_2 deposited under accession number ATCC 98715;
(h) a polynucleotide encoding a protein comprising the amino acid sequence of SEQ ID NO:4;
(i) a polynucleotide encoding a protein comprising a fragment of the amino acid sequence of SEQ ID NO:4, the fragment comprising eight contiguous amino acids of SEQ ID NO:4;
(j) a polynucleotide that hybridizes under conditions at least as stringent as 4X SSC at 65 degrees C, or 4X SSC at 42 degrees C with 50%
formamide, to any one of the polynucleotides specified in (a)-(i); and (k) a polynucleotide that hybridizes under conditions at least as stringent as 4X SSC at 50 degrees C, or 6X SSC at 40 degrees C with 50%
formamide, to any one of the polynucleotides specified in (a)-(i), and that has a length that is at least 25% of the length of SEQ ID NO:3.

13. A protein comprising an amino acid sequence selected from the group consisting of:
(a) the amino acid sequence of SEQ ID NO:4;
(b) a fragment of the amino acid sequence of SEQ ID NO:4, the fragment comprising eight contiguous amino acids of SEQ ID NO:4; and (c) the amino acid sequence encoded by the cDNA insert of clone ct547_2 deposited under accession number ATCC 98715;
the protein being substantially free from other mammalian proteins.

14. An isolated polynucleotide selected from the group consisting of:
(a) a polynucleotide comprising the nucleotide sequence of SEQ ID
NO:5;
(b) a polynucleotide comprising the nucleotide sequence of SEQ ID
NO:5 from nucleotide 206 to nucleotide 415;
(c) a polynucleotide comprising the nucleotide sequence of SEQ ID
NO:5 from nucleotide 293 to nucleotide 415;
(d) a polynucleotide comprising the nucleotide sequence of the full-length protein coding sequence of clone en553_1 deposited under accession number ATCC 98715;
(e) a polynucleotide encoding the full-length protein encoded by the cDNA insert of clone en553_1 deposited under accession number ATCC 98715;
(f) a polynucleotide comprising the nucleotide sequence of the mature protein coding sequence of clone en553_1 deposited under accession number ATCC 98715;
(g) a polynucleotide encoding the mature protein encoded by the cDNA insert of clone en553_1 deposited under accession number ATCC 98715;
(h) a polynucleotide encoding a protein comprising the amino acid sequence of SEQ ID NO:6;
(i) a polynucleotide encoding a protein comprising a fragment of the amino and sequence of SEQ ID NO:6, the fragment comprising eight contiguous amino acids of SEQ ID NO:6;
(j) a polynucleotide that hybridizes under conditions at least as stringent as 4X SSC at 65 degrees C, or 4X SSC at 42 degrees C with 50%
formamide, to any one of the polynucleotides specified in (a)-(i); and (k) a polynucleotide that hybridizes under conditions at least as stringent as 4X SSC at 50 degrees C, or 6X SSC at 40 degrees C with 50%
formamide, to any one of the polynucleotides specified in (a)-(i), and that has a length that is at least 25% of the length of SEQ ID NO:5.

85 25. A protein comprising an amino acid sequence selected from the group consisting of:
(a) the amino acid sequence of SEQ ID NO:6;
(b) a fragment of the amino acid sequence of SEQ ID NO:6, the fragment comprising eight contiguous amino acids of SEQ ID NO:6; and (c) the amino acid sequence encoded by the cDNA insert of clone en553_1 deposited under accession number ATCC 98715;
the protein being substantially free from other mammalian proteins.

16. An isolated polynucleotide selected from the group consisting of:
(a) a polynucleotide comprising the nucleotide sequence of SEQ ID
NO:7;
(b) a polynucleotide comprising the nucleotide sequence of SEQ ID
NO:7 from nucleotide 120 to nucleotide 1202;
(c) a polynucleotide comprising the nucleotide sequence of the full-length protein coding sequence of clone nn296_2 deposited under accession number ATCC 98715;
(d) a polynucleotide encoding the full-length protein encoded by the cDNA insert of clone nn296_2 deposited under accession number ATCC 98715;
(e) a polynucleotide encoding a protein comprising the amino acid sequence of SEQ ID NO:8;
(f) a polynucleotide encoding a protein comprising a fragment of the amino acid sequence of SEQ ID NO:8, the fragment comprising eight contiguous amino acids of SEQ ID NO:8;
(g) a polynucleotide that hybridizes under conditions at least as stringent as 4X SSC at 65 degrees C, or 4X SSC at 42 degrees C with 50%
formamide, to any one of the polynucleotides specified in (a)-(f); and (h) a polynucleotide that hybridizes under conditions at least as stringent as 4X SSC at 50 degrees C, or 6X SSC at 40 degrees C with 50%
formamide, to any one of the polynucleotides specified in (a)-(f), and that has a length that is at least 25% of the length of SEQ ID NO:7.

17. A protein comprising an amino acid sequence selected from the group consisting of:

(a) the amino acid sequence of SEQ ID NO:8;
(b) a fragment of the amino acid sequence of SEQ ID NO:8, the fragment comprising eight contiguous amino acids of SEQ ID NO:8; and (c) the amino acid sequence encoded by the cDNA insert of clone nn296_2 deposited under accession number ATCC 98715;
the protein being substantially free from other mammalian proteins.

18. An isolated polynucleotide selected from the group consisting of:
(a) a polynucleotide comprising the nucleotide sequence of SEQ ID
NO:9;
(b) a polynucleotide comprising the nucleotide sequence of SEQ ID
NO:9 from nucleotide 597 to nucleotide 704;
(c) a polynucleotide comprising the nucleotide sequence of the full-length protein coding sequence of clone nq27_13 deposited under accession number ATCC 98715;
(d) a polynucleotide encoding the full-length protein encoded by the cDNA insert of clone nq27_13 deposited under accession number ATCC 98715;
(e) a polynucleotide encoding a protein comprising the amino acid sequence of SEQ ID NO:10;
(f) a polynudeotide encoding a protein comprising a fragment of the amino and sequence of SEQ ID NO:10, the fragment comprising eight contiguous amino acids of SEQ ID NO:10;
(g) a polynucleotide encoding a protein comprising the amino acid sequence of SEQ ID NO:32;
(h) a polynucleotide encoding a protein comprising a fragment of the amino acid sequence of SEQ ID NO:32, the fragment comprising eight contiguous amino acids of SEQ ID NO:32;
(i) a polynucleotide that hybridizes under conditions at least as stringent as 4X SSC at 65 degrees C, or 4X SSC at 42 degrees C with 50%
formamide, to any one of the polynucleotides specified in (a)-(h); and (j) a polynucleotide that hybridizes under conditions at least as stringent as 4X SSC at 50 degrees C, or 6X SSC at 40 degrees C with 50%
formamide, to any one of the polynucleotides specified in (a)-(h), and that has a length that is at least 25% of the length of SEQ ID NO:9.

19. A protein comprising an amino acid sequence selected from the group consisting of:
(a) the amino acid sequence of SEQ ID NO:10;
(b) a fragment of the amino acid sequence of SEQ ID NO:10, the fragment comprising eight contiguous amino acids of SEQ ID NO:10; and (c) the amino acid sequence of SEQ ID NO:32;
(d) a fragment of the amino acid sequence of SEQ ID NO:32, the fragment comprising eight contiguous amino acids of SEQ ID NO:32; and (e) the amino acid sequence encoded by the cDNA insert of clone nq27_13 deposited under accession number ATCC 98715;
the protein being substantially free from other mammalian proteins.

20. An isolated polynucleotide selected from the group consisting of:
(a) a polynucleotide comprising the nucleotide sequence of SEQ ID
NO:11;
(b) a polynucleotide comprising the nucleotide sequence of SEQ ID
NO:11 from nucleotide 44 to nucleotide 475;
(c) a polynucleotide comprising the nucleotide sequence of the full-length protein coding sequence of clone pk65 4 deposited under accession number ATCC 98715;
(d) a polynucleotide encoding the full-length protein encoded by the cDNA insert of clone pk65 4 deposited under accession number ATCC 98715;
(e) a polynucleotide encoding a protein comprising the amino acid sequence of SEQ ID NO:12;
(f) a polynucleotide encoding a protein comprising a fragment of the amino acid sequence of SEQ ID NO:12, the fragment comprising eight contiguous amino acids of SEQ ID NO:12;
(g) a polynucleotide that hybridizes under conditions at least as stringent as 4X SSC at 65 degrees C, or 4X SSC at 42 degrees C with 50%
formamide, to any one of the polynucleotides specified in (a)-(f); and (h) a polynucleotide that hybridizes under conditions at least as stringent as 4X SSC at 50 degrees C, or 6X SSC at 40 degrees C with 50%
formamide, to any one of the polynucleotides specified in (a)-(f), and that has a length that is at least 25% of the length of SEQ ID NO:11.

21. A protein comprising an amino acid sequence selected from the group consisting of:
(a) the amino acid sequence of SEQ ID NO:12;
(b) a fragment of the amino acid sequence of SEQ ID NO:12, the fragment comprising eight contiguous amino acids of SEQ ID NO:12; and (c) the amino acid sequence encoded by the cDNA insert of clone pk65_4 deposited under accession number ATCC 98715;
the protein being substantially free from other mammalian proteins.

22. An isolated polynucleotide selected from the group consisting of:
(a) a polynucleotide comprising the nucleotide sequence of SEQ ID
NO:13;
(b) a polynucleotide comprising the nucleotide sequence of SEQ ID
NO:13 from nucleotide 285 to nucleotide 590;
(c) a polynucleotide comprising the nucleotide sequence of SEQ ID
NO:13 from nucleotide 408 to nucleotide 590;
(d) a polynucleotide comprising the nucleotide sequence of the full-length protein coding sequence of clone pk855_1 deposited under accession number ATCC 98715;
(e) a polynucleotide encoding the full-length protein encoded by the cDNA insert of clone pk855_1 deposited under accession number ATCC 98715;
(f) a polynucleotide comprising the nucleotide sequence of the mature protein coding sequence of clone pk855_1 deposited under accession number ATCC 98715;
(g) a polynucleotide encoding the mature protein encoded by the cDNA insert of clone pk855_1 deposited under accession number ATCC 98715;
(h) a polynucleotide encoding a protein comprising the amino acid sequence of SEQ ID NO:14;
(i) a polynucleotide encoding a protein comprising a fragment of the amino acid sequence of SEQ ID NO:14, the fragment comprising eight contiguous amino acids of SEQ ID NO:14;
(j) a polynucleotide that hybridizes under conditions at least as stringent as 4X SSC at 65 degrees C, or 4X SSC at 42 degrees C with 50%
formamide, to any one of the polynucleotides specified in (a)-(i); and (k) a polynucleotide that hybridizes under conditions at least as stringent as 4X SSC at 50 degrees C, or 6X SSC at 40 degrees C with 50%
formamide, to any one of the polynucleotides specified in (a)-(i), and that has a length that is at least 25% of the length of SEQ ID NO:13.

23. A protein comprising an amino acid sequence selected from the group consisting of:
(a) the amino acid sequence of SEQ ID NO:14;
(b) a fragment of the amino acid sequence of SEQ ID NO:14, the fragment comprising eight contiguous amino acids of SEQ ID NO:14; and (c) the amino acid sequence encoded by the cDNA insert of clone pk855_1 deposited under accession number ATCC 98715;
the protein being substantially free from other mammalian proteins.

24. An isolated polynucleotide selected from the group consisting of:
(a) a polynucleotide comprising the nucleotide sequence of SEQ ID
NO:15;
(b) a polynucleotide comprising the nucleotide sequence of SEQ ID
NO:15 from nucleotide 193 to nucleotide 2223;
(c) a polynucleotide comprising the nucleotide sequence of SEQ ID
NO:15 from nucleotide 1045 to nucleotide 2223;
(d) a polynucleotide comprising the nucleotide sequence of the full-length protein coding sequence of clone PL776_6 deposited under accession number ATCC 98715;
(e) a polynucleotide encoding the full-length protein encoded by the cDNA insert of clone PL776_6 deposited under accession number ATCC 98715;
(f) a polynucleotide comprising the nucleotide sequence of the mature protein coding sequence of clone PL776_6 deposited under accession number ATCC 98715;
(g) a polynucleotide encoding the mature protein encoded by the cDNA insert of clone PL776_6 deposited under accession number ATCC 98715;
(h) a polynucleotide encoding a protein comprising the amino acid sequence of SEQ ID NO:16;

(i) a polynucleotide encoding a protein comprising a fragment of the amino acid sequence of SEQ ID NO:16, the fragment comprising eight contiguous amino acids of SEQ ID NO:16;
(j) a polynucleotide that hybridizes under conditions at least as stringent as 4X SSC at 65 degrees C, or 4X SSC at 42 degrees C with 50%
formamide, to any one of the polynucleotides specified in (a)-(i); and (k) a polynucleotide that hybridizes under conditions at least as stringent as 4X SSC at 50 degrees C, or 6X SSC at 40 degrees C with 50%
formamide, to any one of the polynucleotides specified in (a)-(i), and that has a length that is at least 25% of the length of SEQ ID NO:15.

25. A protein comprising an amino acid sequence selected from the group consisting of:
(a) the amino acid sequence of SEQ ID NO:16;
(b) a fragment of the amino acid sequence of SEQ ID NO:16, the fragment comprising eight contiguous amino acids of SEQ ID NO:16; and (c) the amino acid sequence encoded by the cDNA insert of clone PL776_6 deposited under accession number ATCC 98715;
the protein being substantially free from other mammalian proteins.

26. An isolated polynucleotide selected from the group consisting of:
(a) a polynucleotide comprising the nucleotide sequence of SEQ ID
NO:17;
(b) a polynucleotide comprising the nucleotide sequence of SEQ ID
NO:17 from nucleotide 198 to nucleotide 1121;
(c) a polynucleotide comprising the nucleotide sequence of SEQ ID
NO:17 from nucleotide 381 to nucleotide 1121;
(d) a polynucleotide comprising the nucleotide sequence of the full-length protein coding sequence of clone pm4_13 deposited under accession number ATCC 98715;
(e) a polynucleotide encoding the full-length protein encoded by the cDNA insert of clone pm4_13 deposited under accession number ATCC 98715;

(f) a polynucleotide comprising the nucleotide sequence of the mature protein coding sequence of clone pm4_13 deposited under accession number ATCC 98715;
(g) a polynucleotide encoding the mature protein encoded by the cDNA insert of clone pm4_13 deposited under accession number ATCC 98715;
(h) a polynucleotide encoding a protein comprising the amino acid sequence of SEQ ID NO:18;
(i) a polynucleotide encoding a protein comprising a fragment of the amino acid sequence of SEQ ID NO:18, the fragment comprising eight contiguous amino acids of SEQ ID NO:18;
(j) a polynucleotide that hybridizes under conditions at least as stringent as 4X SSC at 65 degrees C, or 4X SSC at 42 degrees C with 50%
formamide, to any one of the polynucleotides specified in (a)-(i); and (k) a polynucleotide that hybridizes under conditions at least as stringent as 4X SSC at 50 degrees C, or 6X SSC at 40 degrees C with 50%
formamide, to any one of the polynucleotides specified in (a)-(i), and that has a length that is at least 25% of the length of SEQ ID NO:17.

27. A protein comprising an amino acid sequence selected from the group consisting of:
(a) the amino acid sequence of SEQ ID NO:18;
(b) a fragment of the amino acid sequence of SEQ ID NO:18, the fragment comprising eight contiguous amino acids of SEQ ID NO:18; and (c) the amino acid sequence encoded by the cDNA insert of clone pm4_13 deposited under accession number ATCC 98715;
the protein being substantially free from other mammalian proteins.

28. An isolated polynucleotide selected from the group consisting of:
(a) a polynucleotide comprising the nucleotide sequence of SEQ ID
NO:19;
(b) a polynucleotide comprising the nucleotide sequence of SEQ ID
NO:19 from nucleotide 19 to nucleotide 1953;

(c) a polynucleotide comprising the nucleotide sequence of the full-length protein coding sequence of clone pt326_4 deposited under accession number ATCC 98715;
(d) a polynucleotide encoding the full-length protein encoded by the cDNA insert of clone pt326_4 deposited under accession number ATCC 98715;
(e) a polynucleotide encoding a protein comprising the amino acid sequence of SEQ ID NO:20;
(f) a polynucleotide encoding a protein comprising a fragment of the amino acid sequence of SEQ ID NO:20, the fragment comprising eight contiguous amino acids of SEQ ID NO:20;
(g) a polynucleotide that hybridizes under conditions at least as stringent as 4X SSC at 65 degrees C, or 4X SSC at 42 degrees C with 50%
formamide, to any one of the polynucleotides specified in (a)-(f); and (h) a polynucleotide that hybridizes under conditions at least as stringent as 4X SSC at 50 degrees C, or 6X SSC at 40 degrees C with 50%
formamide, to any one of the polynucleotides specified in (a)-(f), and that has a length that is at least 25% of the length of SEQ ID NO:19.

29. A protein comprising an amino acid sequence selected from the group consisting of:
(a) the amino acid sequence of SEQ ID NO:20;
(b) a fragment of the amino acid sequence of SEQ ID NO:20, the fragment comprising eight contiguous amino acids of SEQ ID NO:20; and (c) the amino acid sequence encoded by the cDNA insert of clone pt326_4 deposited under accession number ATCC 98715;
the protein being substantially free from other mammalian proteins.