CA2317392A1 - Secreted proteins and polynucleotides encoding them - Google Patents

Secreted proteins and polynucleotides encoding them Download PDF

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CA2317392A1
CA2317392A1 CA002317392A CA2317392A CA2317392A1 CA 2317392 A1 CA2317392 A1 CA 2317392A1 CA 002317392 A CA002317392 A CA 002317392A CA 2317392 A CA2317392 A CA 2317392A CA 2317392 A1 CA2317392 A1 CA 2317392A1
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seq
nucleotide
polynucleotide
nucleotide sequence
sequence
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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
Vikki Spaulding
Gordon G. Wong
Hilary F. Clark
Kim Fechtel
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Genetics Institute LLC
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    • C07K14/46—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from vertebrates
    • C07K14/47—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from vertebrates from mammals
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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P7/00—Drugs for disorders of the blood or the extracellular fluid
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
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    • A61K38/00—Medicinal preparations containing peptides

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Abstract

Novel polynucleotides and the proteins encoded thereby are disclosed.

Description

This application is a continuation-in-part of provisional application Ser. No.
60/071,304, filed January 13,1998, which is incorporated by reference herein.

The present invention provides novel polynudeotides and proteins encoded by such polynuBeotides, 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, 2 0 such as lymphokines, interferons, CSFs and interleukins) has matured rapidly over the past decade. The now routine hybridization cloning and expression cloning techniques done novel polynudeotides "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 2 5 Boning). More recent "indirect" Boning techniques such as signal sequence cloning, which isolates DNA sequences based on the presence of a now well-recognized secretory leader 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 3 0 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 polynudeotides encoding them that the present invention is directed.

WO 99!36512 . . PGT/US99/00550 SUN111~IARY OF T 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
N0:1;
(b) a polynucleotide comprising the nucleotide sequence of SEQ ID
NO:1 from nucleotide 427 to nucleotide 1146;
(c) a polynucleotide comprising the nucleotide sequence of SEQ ID
NO:1 from nucleotide 589 to nucleotide 1146;
(d) a polynucleotide comprising the nucleotide sequence of the full-length protein coding sequence of clone bd577 1 deposited under accession number ATCC 98631;
(e) a polynucleotide encoding the full-length protein encoded by the cDNA insert of clone bd577 1 deposited under accession number ATCC 98631;
(f) a polynucleotide comprising the nucleotide sequence of a mature protein coding sequence of clone bd577 1 deposited under accession number ATCC 98631;
(g) a polynucleotide encoding a mature protein encoded by the cDNA
insert of clone bd5~ 1 deposited under accession number ATCC 98631;
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 consecutive amino acids of SEQ ID N0:2;
2 5 (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 ; and (1) a polynucleotide that hybridizes under stringent conditions to any 3 0 one of the polynucleotides specified in (a)-(i).
Preferably, such polynucleotide comprises the nucleotide sequence of SEQ ID
NO:1 from nucleotide 427 to nucleotide 1146; the nucleotide sequence of SEQ ID
N0:1 from nucleotide 589 to nucleotide 1146; the nucleotide sequence of the full-length protein coding sequence of clone bd577 1 deposited under accession number ATCC 98631;
or the V1~0 99/3b512 PCTNS99/00550 nucleotide sequence of a mature protein coding sequence of clone bd5~ 1 deposited under accession number ATCC 98631. In other preferred embodiments, the polynueleotide encodes the full-length or a mature protein encoded by the cDNA
insert of clone bd577 1 deposited under accession number ATCC 98631. 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) consecutive 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 115 to amino acid 124 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 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 NO:1, but excluding the poly(A) tail at the 3' end of SEQ ID N0:1; and (ab) the nucleotide sequence of the cDNA insert of clone bd5~ 1 deposited under accession number ATCC 98631;
(ii) hybridizing said probes) to human genomic DNA in 2 5 conditions at least as stringent as 4X SSC at 65 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 NO:1, but excluding the poly(A) tail at the 3' end of SEQ ID NO:1; and (bb) the nucleotide sequence of the cDNA insert of clone bd577 1 deposited under accession number ATCC 98631;
(ii) hybridizing said primers) to human genomic DNA in conditions at least as stringent as 4X SSC at 65 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: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 N0:1, but excluding the poly(A) tail at the 3' end of SEQ ID N0:1. 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 427 to nucleotide 1146, and extending contiguously from a nucleotide sequence corresponding to the 5' end of said sequence of SEQ ID NO:1 from nucleotide 427 to nucleotide 1146, to a nucleotide sequence corresponding to the 3' end of said sequence of SEQ ID NO:1 from nucleotide 427 to nucleotide 1146. 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 589 to nucleotide 1146, and extending contiguously from a 2 0 nucleotide sequence corresponding to the 5' end of said sequence of SEQ ID
NO:1 from nucleotide 589 to nucleotide 1146, to a nucleotide sequence corresponding to the 3' end of said sequence of SEQ ID NO:1 from nucleotide 589 to nucleotide 1146.
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 N0:2;
(b) fragments of the amino acid sequence of SEQ ID N0:2, each fragment comprising-eight consecutive amino acids of SEQ ID N0:2; and (c) the amino acid sequence encoded by the cDNA insert of clone 3 0 bd577 1 deposited under accession number ATCC 98631;
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) consecutive 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 115 to amino acid 124 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 polynucleotide comprising the nucleotide sequence of SEQ ID
N0:3;
(b) a polynudeotide comprising the nucleotide sequence of SEQ ID
N0:3 from nucleotide 95 to nucleotide 1522;
(c) a polynudeotide comprising the nucleotide sequence of SEQ ID
N0:3 from nucleotide 161 to nucleotide 1522;
(d) a polynucleotide comprising the nucleotide sequence of the full-length protein coding sequence of clone bv280_3 deposited under accession number ATCC 98631;
(e) a polynudeotide encoding the full-length protein encoded by the cDNA insert of clone bv280_3 deposited under accession number ATCC 98631;
(f) a polynucleotide comprising the nucleotide sequence of a mature protein coding sequence of done bv280_3 deposited under accession number 2 0 ATCC 98631;
(g) a polynudeotide encoding a mature protein encoded by the cDNA
insert of clone bv280 3 deposited under accession number ATCC 98631;
(h) a polynucleotide encoding a protein comprising the amino acid sequence of SEQ ID N0:4;
2 5 (i) a polynudeotide encoding a protein comprising a fragment of the amino acid sequence of SEQ ID N0:4 having biological activity, the fragment comprising eight consecutive amino acids of SEQ ID N0:4;
(j) a polynudeotide 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 ; and (1) a polynucleotide that hybridizes under stringent conditions to any one of the polynucleotides specified in (a)-(i).
Preferably, such polynucleotide comprises the nucleotide sequence of SEQ ID
N0:3 from nucleotide 95 to nucleotide 1522; the nucleotide sequence of SEQ ID
N0:3 from nucleotide 161 to nucleotide 1522; the nucleotide sequence of the full-length protein coding sequence of clone bv280_3 deposited under accession number ATCC 98631;
or the nucleotide sequence of a mature protein coding sequence of clone bv280_3 deposited under accession number ATCC 98631. In other preferred embodiments, the polynucleotide codes the full-length or a mature protein encoded by the cDNA
insert of clone bv280_3 deposited under accession number ATCC 98631. 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 activity, the fragment preferably comprising eight (more preferably twenty, most preferably thirty) consecutive amino acids of SEQ ID NO: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 233 to amino acid 242 of SEQ ID N0:4.
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:
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 N0:3, but excluding the poly(A) tail at the 2 5 3' end of SEQ ID N0:3; and (ab) the nucleotide sequence of the cDNA insert of clone bv280 3 deposited under accession number ATCC 98631;
(ii) hybridizing said probes) to human genomic DNA in -conditions at least as stringent as 4X SSC at 65 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 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 bv280 3 deposited under accession number ATCC 98631;
(ii) hybridizing said primers) to human genomic DNA in conditions at least as stringent as 4X SSC at 65 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 N0:3, 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 according to the above process comprises a nucleotide sequence corresponding to the cDNA sequence of SEQ ID N0:3 from nucleotide 95 to nucleotide 1522, and extending contiguously from a nucleotide sequence corresponding to the 5' end of said sequence of 2 0 SEQ ID NO:3 from nucleotide 95 to nucleotide 1522, to a nucleotide sequence corresponding to the 3' end of said sequence of SEQ ID N0:3 from nucleotide 95 to nucleotide 1522. 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 161 to nucleotide 1522, and- extending contiguously from a 2 5 nucleotide sequence corresponding to the 5' end of said sequence of SEQ ID
N0:3 from nucleotide 161 to nucleotide 1522, to a nucleotide sequence corresponding to the 3' end of said sequence of SEQ ID N0:3 from nucleotide 161 to nucleotide 1522.
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 N0:4;
(b) fragments of the amino acid sequence of SEQ ID N0:4, each fragment comprising eight consecutive amino acids of SEQ ID N0:4; and WO 99/36512 P~/US99/00550 (c) the amino acid sequence encoded by the cDNA insert of clone bv280_3 deposited under accession number ATCC 98631;
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) consecutive 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 233 to amino acid 242 of SEQ ID N0:4.
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
N0:5;
(b) a polynucleotide comprising the nucleotide sequence of SEQ ID
N0:5 from nucleotide 286 to nucleotide 552;
(c) a polynucleotide comprising the nucleotide sequence of SEQ ID
N0:5 from nucleotide 475 to nucleotide 552;
(d) a polynucleotide comprising the nucleotide sequence of the full-2 0 length protein coding sequence of clone co315_3 deposited under accession number ATCC 98631;
(e) a polynucleotide encoding the full-length protein encoded by the cDNA insert of clone co315_3 deposited under accession number ATCC 98631;
(f) a polynucleotide comprising the nucleotide sequence of a mature 2 5 protein coding sequence of clone co315 3 deposited under accession number ATCC 98631;
(g) a polynucleotide encoding a mature protein encoded by the cDNA
insert of clone co315_3 deposited under accession number ATCC 98631;
(h) a polynucleotide encoding a protein comprising the amino acid 3 0 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 consecutive amino acids of SEQ ID N0:6;
(j) a palynucleotide which is an allelic variant of a polynucleotide of (a)-(g) above;
{k) a polynudeotide which encodes a species homologue of the protein of (h) or (i) above ; and (1) a polynucleotide that hybridizes under stringent conditions to any one of the polynucleotides specified in (ar(i).
Preferably, such polynucleotide comprises the nucleotide sequence of SEQ ID
N0:5 from nucleotide 286 to nucleotide 552; the nucleotide sequence of SEQ ID
N0:5 from nucleotide 475 to nucleotide 552; the nucleotide sequence of the full-length protein coding sequence of clone co315_3 deposited under accession number ATCC 98631; or the nucleotide sequence of a mature protein coding sequence of clone co315_3 deposited under accession number ATCC 98631. In other preferred embodiments, the polynucleotide encodes the full-length or a mature protein encoded by the cDNA
insert of clone co315_3 deposited under accession number ATCC 98631. In further preferred embodiments, the present invention provides a polynucleotide encoding a protein comprising a fragment of the amino acid sequence of SEQ ID N0:6 having biological activity, the fragment preferably comprising eight (more preferably twenty, most preferably thirty) consecutive amino acids of SEQ ID N0:6, or a polynucleoiide encoding a protein comprising a fragment of the amino acid sequence of SEQ ID N0:6 having 2 0 biological activity, the fragment comprising the amino acid sequence from amino acid 39 to amino acid 48 of SEQ ID N0:6.
Other embodiments provide the gene corresponding,to the cDNA sequence of SEQ
ID N0:5.
Further embodiments of the invention provide isolated polynucleotides produced 2 5 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:
3 0 (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 co315_3 deposited under accession number ATCC 98631;
(ii) hybridizing said probes) to human genomic DNA in conditions at least as stringent as 4X SSC at 65 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 N0:5, 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 co315_3 deposited under accession number ATCC 98631;
(ii) hybridizing said primers) to human genomic DNA in conditions at least as stringent as 4X SSC at 65 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 2 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 286 to nucleotide 552, and extending contiguously from a nucleotide sequence corresponding to the 5' end of said sequence of SEQ ID N0:5 from nucleotide 286 to nucleotide 552, to a nucleotide sequence corresponding to the 3' end of said sequence of SEQ ID N0:5 from nucleotide 286 to nucleotide 552. Also preferably the polynucleotide isolated according to the above process comprises a nucleotide sequence corresponding to the cDNA sequence of SEQ ID
3 0 N0:5 from nucleotide 475 to nucleotide 552, and extending contiguously from a nucleotide sequence corresponding to the 5' end of said sequence of SEQ ID
N0:5 from nucleotide 475 to nucleotide 552, to a nucleotide sequence corresponding to the 3' end of said sequence of SEQ ID N0:5 from nucleotide 475 to nucleotide 552.

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) fragments of the amino acid sequence of SEQ ID N0:6, each fragment comprising eight consecutive amino acids of SEQ ID N0:6; and (c) the amino acid sequence encoded by the cDNA insert of clone co315_3 deposited under accession number ATCC 98631;
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 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) consecutive amino acids 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 amino acid 39 to amino acid 48 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
2 0 N0:7;
(b) a polynucleotide comprising the nucleotide sequence of SEQ ID
N0:7 from nucleotide 1682 to nucleotide 1963;
(c) a polynucleotide comprising the nucleotide sequence of the full-length protein coding sequence of clone ij226_6 deposited under accession number 2 5 ATCC 98631;
(d) a polynucleotide encoding the full-length protein encoded by the cDNA insert of clone ij226_6 deposited under accession number ATCC 98631;
(e) a polynucleotide comprising the nucleotide sequence of a mature protein coding sequence of clone ij226_6 deposited under accession number ATCC
3 0 98631;
(f) a polynucleotide encoding a mature protein encoded by the cDNA
insert of clone ij226_6 deposited under accession number ATCC 98631;
(g) a polynucleotide encoding a protein comprising the amino acid sequence of SEQ ID N0:8;

WO 99/36512 PCfIUS99l00550 (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 consecutive 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 ; and (k) a polynucleotide that hybridizes under stringent conditions to any one of the polynucleotides specified in (a)-(h).
Preferably, such polynucleotide comprises the nucleotide sequence of SEQ ID
N0:7 from nucleotide 1682 to nucleotide 1963; the nucleotide sequence of the full-length protein coding sequence of clone ij226_6 deposited under accession number ATCC
98631;
or the nucleotide sequence of a mature protein coding sequence of clone ij226_6 deposited under accession number ATCC 98631. In other preferred embodiments, the polynucleotide encodes the full-length or a mature protein encoded by the cDNA
insert of clone ij226 6 deposited under accession number ATCC 98631. In further preferred embodiments, the present invention provides a polynucleotide encoding a protein comprising a fragment of the amino acid sequence of SEQ ID N0:8 having biological activity, the fragment preferably comprising eight (more preferably twenty, most 2 0 preferably thirty) consecutive 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 acid 42 to amino acid 51 of SEQ ID N0:8.
Other embodiments provide the gene corresponding to the cDNA sequence of SEQ
2 5 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 polynudeotide probes that hybridize 3 0 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 WO 99!36512 PCT/US99/00550 (ab) the nucleotide sequence of the cDNA insert of clone ij226 6 deposited under accession number ATCC 98631;
(ii) hybridizing said probes) to human genomic DNA in conditions at least as stringent as 4X SSC at 65 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 N0:7, but excluding the poly(A) tail at the 3' end of SEQ ID N0:7; and (bb) the nucleotide sequence of the cDNA insert of clone ij226_6 deposited under accession number ATCC 98631;
(ii) hybridizing said primers) to human genomic DNA in conditions at least as stringent as 4X SSC at 65 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 N0:7, 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 2 5 according to the above process comprises a nucleotide sequence corresponding to the cDNA sequence of SEQ ID N0:7 from nucleotide 1682 to nucleotide 1963, and extending contiguously from a nucleotide sequence corresponding to the 5' end of said sequence of SEQ ID N0:7 from nucleotide 1682 to nucleotide 1963, to a nucleotide sequence corresponding to the 3' end of said sequence of SEQ ID N0:7 from nucleotide 1682 to 3 0 nucleotide 1963.
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) fragments of the amino acid sequence of SEQ ID N0:8, each fragment comprising eight consecutive amino acids of SEQ ID N0:8; and (c) the amino acid sequence encoded by the cDNA insert of clone ij226_6 deposited under accession number ATCC 98631;
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 amino acid sequence of SEQ ID N0:8 having biological activity, the fragment preferably comprising eight (more preferably twenty, most preferably thirty) consecutive amino acids 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 42 to amino acid 51 of SEQ ID N0:8.
In one embodiment, the present invention provides a composition comprising an isolated polynucleotide selected from the group consisting of:
1.5 (a) a polynucleotide comprising the nucleotide sequence of SEQ ID
N0:9;
(b) a polynucleotide comprising the nucleotide sequence of SEQ ID
N0:9 from nucleotide 1137 to nucleotide 1346;
(c) a polynucleotide comprising the nucleotide sequence of the full-2 0 length protein coding sequence of clone nf443_1 deposited under accession number ATCC 98631;
(d) a polynucleotide encoding the full-length protein encoded by the cDNA insert of clone nf4~_1 deposited under accession number ATCC 98631;
(e) a polynucleotide comprising the nucleotide sequence of a mature 2 5 protein coding sequence of clone nf443_1 deposited under accession number ATCC 98631;
(f) a polynucleotide encoding a mature protein encoded by the cDNA
insert of clone nf443_1 deposited under accession number ATCC 98631;
(g) a polynucleotide encoding a protein comprising the amino acid 3 0 sequence of SEQ ID NO: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 consecutive amino acids of SEQ ID N0:10;

(i) a polynucleotide which is an allelic variant of a polynucleotide of (a)-(f) above;
(j) a polynucleotide which encodes a speaes homologue of the protein of (g) or (h) above ; and (k) a polynucleotide that hybridizes under stringent conditions to any one of the polynucleotides specified in (a)-(h).
Preferably, such polynucleotide comprises the nucleotide sequence of SEQ ID
N0:9 from nucleotide 1137 to nucleotide 1346; the nucleotide sequence of the full-length protein coding sequence of clone nf443_1 deposited under accession number ATCC
98631;
or the nucleotide sequence of a mature protein coding sequence of clone nf443_1 deposited under accession number ATCC 98631. In other preferred embodiments, the polynucleotide encodes the full-length or a mature protein encoded by the cDNA
insert of clone nf443_1 deposited under accession number ATCC 98631. 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) consecutive amino acids of SEQ ID N0:10, or a polynucleotide encoding a protein comprising a fragment of the amino acid sequence of SEQ ID N0:10 having biological activity, the fragment comprising the amino acid sequence from amino acid 30 2 0 to amino acid 39 of SEQ ID N0:10.
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 0 3' end of SEQ ID N0:9; and (ab) the nucleotide sequence of the cDNA insert of clone nf443_1 deposited under accession number ATCC 98631;
(ii) hybridizing said probes) to human genomic DNA in conditions at least as stringent as 4X SSC at 65 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 N0:9, but excluding the poly{A) tail at the 3' end of SEQ ID N0:9; and (bb) the nucleotide sequence of the cDNA insert of clone nf443_1 deposited under accession number ATCC 98631;
(ii) hybridizing said primers) to human genomic DNA in conditions at least as stringent as 4X SSC at 65 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
NO:9 to a nucleotide sequence corresponding to the 3' end of SEQ ID N0:9 , but excluding the 2 0 poly(A) tail at the 3' end of SEQ ID N0:9. 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 1137 to nucleotide 1346, and extending contiguously from a nucleotide sequence corresponding to the 5' end of said sequence of SEQ ID N0:9 from nucleotide 1137 to nucleotide 1346, to a nucleotide sequence 2 5 corresponding to the 3' end of said sequence of SEQ ID N0:9 from nucleotide 1137 to nucleotide 1346.
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:
3 0 (a) the amino acid sequence of SEQ ID N0:10;
(b) fragments of the amino acid sequence of SEQ ID NO:10, each fragment comprising eight consecutive amino acids of SEQ ID NO:10; and (c) the amino acid sequence encoded by the cDNA insert of clone nf443_1 deposited under accession number ATCC 98631;

the protein being substantially free from other mammalian proteins. Preferably such 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) consecutive amino acids of SEQ ID N0:10, or 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 30 to amino acid 39 of SEQ ID N0:10.
In one embodiment, the present invention provides a composition comprising an isolated polynudeotide selected from the group consisting of:
(a) a polynudeotide comprising the nucleotide sequence of SEQ ID
NO:11;
(b) a polynudeotide comprising the nucleotide sequence of SEQ ID
NO:11 from nucleotide 308 to nucleotide 634;
(c) a polynudeotide comprising the nucleotide sequence of the full-length protein coding sequence of done nt429_1 deposited under accession number ATCC 98631;
(d) a polynudeotide encoding the full-length protein encoded by the cDNA insert of clone nt429_1 deposited under accession number ATCC 98631;
2 0 (e) a polynudeotide comprising the nucleotide sequence of a mature protein coding sequence of clone nt429_1 deposited under accession number ATCC 98631;
(f) a polynudeotide encoding a mature protein encoded by the cDNA
insert of clone nt429_1 deposited under accession number ATCC 98631;
2 5 (g) a polynudeotide encoding a protein comprising the amino acid sequence of SEQ ID N0:12;
(h) a polynudeotide encoding a protein comprising a fragment of the amino acid sequence of SEQ ID N0:12 having biological activity, the fragment comprising eight consecutive amino acids of SEQ ID N0:12;
3 0 (i) a polynudeotide which is an allelic variant of a polynudeotide of (a)-(f) above;
(j) a polynudeotide which encodes a species homologue of the protein of (g) or (h) above ; and (k) a polynucleotide that hybridizes under stringent conditions to any one of the polynucleotides specified in (a)-(h).
Preferably, such polynucleotide comprises the nucleotide sequence of SEQ ID
N0:11 from nucleotide 308 to nucleotide 634; the nucleotide sequence of the full-length protein coding sequence of clone nt429_1 deposited under accession number ATCC
98631;
or the nucleotide sequence of a mature protein coding sequence of clone nt429_1 deposited under accession number ATCC 98631. In other preferred embodiments, the polynucleotide encodes the full-length or a mature protein encoded by the cDNA
insert of clone nt429_1 deposited under accession number ATCC 98631. In yet other preferred embodiments, the present invention provides a polynucleotide encoding a protein comprising the amino acid sequence of SEQ ID N0:12 from amino acid 1 to amino acid 47. 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) consecutive amino acids of SEQ ID N0:12, 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 49 to amino acid 58 of SEQ ID N0:12.
Other embodiments provide the gene corresponding to the cDNA sequence of SEQ
2 0 ID N0:11.
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 2 5 in 6X SSC at 65 degrees C to a nucleotide sequence selected from the group consisting of:
(aa) SEQ ID NO:11, but excluding the poly(A) tail at the 3' end of SEQ ID N0:11; and (ab) the nucleotide sequence of the cDNA insert of clone 3 0 nt429_1 deposited under accession number ATCC 98631;
(ii) hybridizing said probes) to human genomic DNA in conditions at least as stringent as 4X SSC at 65 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 N0:11, 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 nt429_1 deposited under accession number ATCC 98631;
(ii) hybridizing said primers) to human genomic DNA in conditions at least as stringent as 4X SSC at 65 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:I1, 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 N0:11 , 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 2 0 corresponding to the cDNA sequence of SEQ ID N0:11 from nucleotide 308 to nucleotide 634, and extending contiguously from a nucleotide sequence corresponding to the 5' end of said sequence of SEQ ID NO:11 from nucleotide 308 to nucleotide 634, to a nucleotide sequence corresponding to the 3' end of said sequence of SEQ ID N0:11 from nucleotide 308 to nucleotide 634.
2 5 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;
(b) the amino acid sequence of SEQ ID N0:12 from amino acid 1 to 3 0 amino acid 47;
(c) fragments of the amino acid sequence of SEQ ID N0:12, each fragment comprising eight consecutive amino acids of SEQ ID N0:12; and (d) the amino acid sequence encoded by the cDNA insert of clone nt429_1 deposited under accession number ATCC 98631;

the protein being substantially free from other mammalian proteins. Preferably such protein comprises the amino acid sequence of SEQ ID N0:12 or the amino acid sequence of SEQ ID N0:12 from amino acid 1 to amino acid 47. 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} consecutive amino acids of SEQ ID
N0:12, or 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 49 to amino acid 58 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 104 to nucleotide 652;
(c) a polynucleotide comprising the nucleotide sequence of SEQ ID
N0:13 from nucleotide 377 to nucleotide 652;
(d) a polynucleotide comprising the nucleotide sequence of the full-length protein coding sequence of clone pe503_1 deposited under accession 2 0 number ATCC 98631;
(e) a polynucleotide encoding the full-length protein encoded by the cDNA insert of clone pe503_1 deposited under accession number ATCC 98631;
(f) a polynucleotide comprising the nucleotide sequence of a mature protein coding sequence of clone pe503_1 deposited under accession number 2 5 ATCC 98631;
(g) a polynucleotide encoding a mature protein encoded by the cDNA
insert of clone pe503_1 deposited under accession number ATCC 98631;
(h) a polynucleotide encoding a protein comprising the amino acid sequence of SEQ ID N0:14;
3 0 (i) a polynucleotide encoding a protein comprising a fragment of the amino acid sequence of SEQ ID N0:14 having biological activity, the fragment comprising eight consecutive 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 of {h) or (i) above ; and (1) a polynudeotide that hybridizes under stringent conditions to any one of the polynucleotides specified in (a~(i).
Preferably, such polynucleotide comprises the nucleotide sequence of SEQ ID
N0:13 from nucleotide 104 to nucleotide 652; the nucleotide sequence of SEQ ID
N0:13 from nucleotide 377 to nucleotide 652; the nucleotide sequence of the full-length protein coding sequence of clone pe503_1 deposited under accession number ATCC 98631;
or the nucleotide sequence of a mature protein coding sequence of clone pe503_1 deposited under accession number ATCC 98631. In other preferred embodiments, the polynucleotide encodes the full-length or a mature protein encoded by the cDNA
insert of clone pe503_1 deposited under accession number ATCC 98631. In yet other preferred embodiments, the present invention provides a polynucleotide encoding a protein comprising the amino acid sequence of SEQ TD N0:14 from amino acid 69 to amino acid 125. 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) consecutive amino acids of SEQ ID N0:14, or a polynucleotide encoding a protein comprising a fragment of the amino acid sequence of 2 0 SEQ ID N0:14 having biological activity, the fragment comprising the amino acid sequence from amino acid 86 to amino acid 95 of SEQ ID NO:I4.
Other embodiments provide the gene corresponding to the cDNA sequence of SEQ
ID N0:13.
Further embodiments of the invention provide isolated polynucleotides produced 2 5 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:
3 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 pe503_1 deposited under accession number ATCC 98631;

(ii) hybridizing said probes) to human genomic DNA in conditions at least as stringent as 4X SSC at 65 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 N0:13, 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 pe503_1 deposited under accession number ATCC 98631;
(ii) hybridizing said primers) to human genomic DNA in conditions at least as stringent as 4X SSC at 65 degrees C;
(iii) amplifying human DNA sequences; and (iv) isolating the polynucleoHde 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 2 0 extending contiguously from a nucleotide sequence corresponding to the 5' end of SEQ
ID NO:I3 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 104 to nucleotide 2 5 652, and extending contiguously from a nucleotide sequence corresponding to the 5' end of said sequence of SEQ ID N0:13 from nucleotide 104 to nucleotide 652, to a nucleotide sequence corresponding to the 3' end of said sequence of SEQ ID N0:13 from nucleotide 104 to nucleotide 652. Also preferably the polynucleotide isolated according to the above process comprises a nucleotide sequence corresponding to the cDNA sequence of SEQ ID
3 0 N0:13 from nucleotide 377 to nucleotide 652, and extending contiguously from a nucleotide sequence corresponding to the 5' end of said sequence of SEQ ID
N0:13 from nucleotide 377 to nucleotide 652, to a nucleotide sequence corresponding to the 3' end of said sequence of SEQ ID N0:13 from nucleotide 377 to nucleotide 652.

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 N0:14;
(b} the amino acid sequence of SEQ ID N0:14 from amino acid 69 to amino acid I25;
(c) fragments of the amino acid sequence of SEQ ID N0:14, each fragment comprising eight consecutive amino acids of SEQ ID N0:14; and (d) the amino acid sequence encoded by the cDNA insert of clone pe503_1 deposited under accession number ATCC 98631;
the protein being substantially free from other mammalian proteins. Preferably such protein comprises the amino acid sequence of SEQ ID N0:14 or the amino acid sequence of SEQ ID N0:14 from amino acid 69 to amino acid 125. 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) consecutive 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 86 to amino acid 95 of SEQ ID N0:14.
2 0 In one embodiment, the present invention provides a composition comprising an isolated polynudeotide selected from the group consisting of:
(a) a polynucleotide comprising the nucleotide sequence of SEQ ID
N0:15;
(b) a polynudeotide comprising the nucleotide sequence of SEQ ID
2 5 N0:15 from nucleotide 23 to nucleotide 442;
(c) a polynucleotide comprising the nucleotide sequence of SEQ ID
N0:15 from nucleotide 224 to nucleotide 442;
(d) a polynucleotide comprising the nucleotide sequence of the full-length protein coding sequence of done pe834 6 deposited under accession 3 0 number ATCC 98631;
(e) a polynucleotide encoding the full-length protein encoded by the cDNA insert of clone pe834_6 deposited under accession number ATCC 98631;

WO 99136512 PCTIUS99l00550 a polynucleotide comprising the nucleotide sequence of a mature protein coding sequence of clone pe834_6 deposited under accession number ATCC 98631;
(g) a polynucleotide encoding a mature protein encoded by the cDNA
insert of clone pe834_6 deposited under accession number ATCC 98631;
(h) a polynucleotide encoding a protein comprising the amino acid sequence of SEQ ID N0:16;
(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 consecutive amino ands of SEQ ID N0:16;
(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 ; and (1} a polynucleotide that hybridizes under stringent conditions to any one of the polynucleotides specified in (a)-(i).
Preferably, such polynucleotide comprises the nucleotide sequence of SEQ ID
N0:15 from nucleotide 23 to nucleotide 442; the nucleotide sequence of SEQ ID
N0:15 from nucleotide 224 to nucleotide 442; the nucleotide sequence of the full-length protein 2 0 coding sequence of clone peg 6 deposited under accession number ATCC
98631; or the nucleotide sequence of a mature protein coding sequence of clone pe834_6 deposited under accession number ATCC 98631. In other preferred embodiments, the polynucleotide encodes the full-length or a mature protein encoded by the cDNA
insert of clone pe834_6 deposited under accession number ATCC 98631. 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 N0:16 having biological activity, the fragment preferably comprising eight {more preferably twenty, most preferably thirty) consecutive amino acids of SEQ ID N0:16, or a polynudeotide encoding a protein comprising a fragment of the amino acid sequence of SEQ ID N0:16 having 3 0 biological activity, the fragment rnmprising the amino acid sequence from amino acid 65 to amino acid 74 of SEQ ID N0:16.
Other embodiments provide the gene corresponding to the cDNA sequence of SEQ
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 in 6X SSC at 65 degrees C to a nucleotide sequence selected from the group consisting of:
(aa) SEQ ID N0:15, but excluding the poly(A) tail at the 3' end of SEQ ID N0:15; and (ab) the nucleotide sequence of the cDNA insert of clone pe834_6 deposited under accession number ATCC 98631;
(ii) hybridizing said probes) to human genomic DNA in conditions at least as stringent as 4X SSC at 65 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 N0:15, but excluding the poly(A) tail at the 3' end of SEQ ID N0:15; and (bb} the nucleotide sequence of the cDNA insert of clone pe834 6 deposited under accession number ATCC 98631;
(ii) hybridizing said primers) to human genomic DNA in conditions at least as stringent as 4X SSC at 65 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 3 0 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 , but excluding the poly(A) tail at 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 23 to nucleotide 442, and extending contiguously from a nucleotide sequence corresponding to the 5' end of said sequence of SEQ ID N0:15 from nucleotide 23 to nucleotide 442, to a nucleotide sequence corresponding to the 3' end of said sequence of SEQ ID N0:15 from nucleotide 23 to nucleotide 442.
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 N0:16;
(b) fragments of the amino acid sequence of SEQ ID N0:16, each fragment comprising eight consecutive amino acids of SEQ ID N0:16; and (c) the amino acid sequence encoded by the cDNA insert of clone pe834 6 deposited under accession number ATCC 98631;
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) consecutive 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 2 0 sequence from amino acid 65 to amino acid 74 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;
2 5 (b) a polynucleotide comprising the nucleotide sequence of SEQ ID
N0:17 from nucleotide 98 to nucleotide 265;
(c) a polynucleotide comprising the nucleotide sequence of SEQ ID
N0:17 from nucleotide 152 to nucleotide 265;
(d) a polynucleotide comprising the nucleotide sequence of the full-3 0 length protein coding sequence of clone yal0_1 deposited under accession number ATCC 98631;
(e) a polynucleotide encoding the full-length protein encoded by the cDNA insert of clone yal0_1 deposited under accession number ATCC 98631;

(f) a polynucleotide comprising the nucleotide sequence of a mature protein coding sequence of clone yal0_1 deposited under accession number ATCC
98631;
(g) a polynudeotide encoding a mature protein encoded by the cDNA
insert of clone yal0_1 deposited under accession number ATCC 98631;
(h) a polynucleotide encoding a protein comprising the amino acid 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 consecutive 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 ; and (1) a polynucleotide that hybridizes under stringent conditions to any one of the polynucleotides specified in (a~(i).
Preferably, such polynucleotide comprises the nucleotide sequence of SEQ ID
N0:17 from nucleotide 98 to nucleotide 265; the nucleotide sequence of SEQ ID
N0:17 from nucleotide 152 to nucleotide 265; the nucleotide sequence of the full-length protein 2 0 coding sequence of clone yal0_1 deposited under accession number ATCC
98631; or the nucleotide sequence of a mature protein coding sequence of clone yal0_1 deposited under accession number ATCC 98631. In other preferred embodiments, the polynucleotide encodes the full-length or a mature protein encoded by the cDNA insert of clone yal0_1 deposited under accession number ATCC 98631. In further preferred embodiments, the 2 5 present invention provides a polynucleotide encoding a protein comprising a fragment of the amino acid sequence of SEQ ID N0:18 having biological activity, the fragment preferably rnmprising eight (more preferably twenty, most preferably thirty) consecutive amino acids 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 3 0 fragment comprising the amino acid sequence from amino acid 22 to amino acid 31 of SEQ
ID N0:18.
Other embodiments provide the gene corresponding to the cDNA sequence of SEQ
ID N0:17.

WO 99/36512 PCfIUS99/00550 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 N0:17, 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 yal0_1 deposited under accession number ATCC 98631;
(ii) hybridizing said probes) to human genomic DNA in conditions at least as stringent as 4X SSC at 65 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 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 yal0_1 deposited under accession number ATCC 98631;
(ii) hybridizing said primers) to human genomic DNA in 2 5 conditions at least as stringent as 4X SSC at 65 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 N0:17, and 3 0 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 NO:17 , but excluding the poly(A) tail at the 3' end of SEQ ID N0:17. 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 98 to nucleotide 265, and extending contiguously from a nucleotide sequence corresponding to the 5' end of said sequence of SEQ ID N0:17 from nucleotide 98 to nucleotide 265, to a nucleotide sequence corresponding to the 3' end of said sequence of SEQ ID N0:17 from nucleotide 98 to nucleotide 265. 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 152 to nucleotide 265, and extending contiguously from a nucleotide sequence corresponding to the 5' end of said sequence of SEQ ID
N0:17 from nucleotide 152 to nucleotide 265, to a nucleotide sequence corresponding to the 3' end of said sequence of SEQ ID N0:17 from nucleotide 152 to nucleotide 265.
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) fragments of the amino acid sequence of SEQ ID N0:18, each fragment comprising eight consecutive amino acids of SEQ ID N0:18; and (c) the amino acid sequence encoded by the cDNA insert of clone yal0_1 deposited under accession number ATCC 98631;
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 2 0 embodiments, the present invention provides a protein comprising a fragment of the amino acid sequence of SEQ ID N0:18 having biological activity, the fragment preferably comprising eight (more preferably twenty, most preferably thirty) consecutive 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 2 5 sequence from amino acid 22 to amino acid 31 of SEQ ID N0:18.
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
N0:19;
3 0 (b) a polynucleotide comprising the nucleotide sequence of SEQ ID
N0:19 from nucleotide 176 to nucleotide 583;
(c) a polynucleotide comprising the nucleotide sequence of the full-length protein coding sequence of clone yb40_1 deposited under accession number ATCC 98631;

(d) a polynudeotide encoding the full-length protein encoded by the cDNA insert of clone yb40 1 deposited under accession number ATCC 98631;
(e) a polynudeotide comprising the nucleotide sequence of a mature protein coding sequence of done yb40_1 deposited under accession number ATCC
98631;
a polynudeotide encoding a mature protein encoded by the cDNA
insert of clone yb40 1 deposited under accession number ATCC 98631;
(g) a polynudeotide encoding a protein comprising the amino acid sequence of SEQ ID N0:20;
(h) a polynudeotide encoding a protein comprising a fragment of the amino acid sequence of SEQ ID N0:20 having biological activity, the fragment comprising eight consecutive amino acids of SEQ ID N0:20;
(i) a polynudeotide which is an allelic variant of a polynudeotide of (a)-(f) above;
(j) a polynudeotide which encodes a species homologue of the protein of (g) or (h) above ; and a polynudeotide that hybridizes under stringent conditions to any one of the polynudeotides specified in (a)-(h).
Preferably, such polynudeotide comprises the nucleotide sequence of SEQ ID
2 0 N0:19 from nucleotide 176 to nucleotide 583; the nucleotide sequence of the full-length protein coding sequence of clone yb40_1 deposited under accession number ATCC
98631;
or the nucleotide sequence of a mature protein coding sequence of clone yb40_1 deposited under accession number ATCC 98631. In other preferred embodiments, the polynudeotide encodes the full-length or a mature protein encoded by the cDNA
insert 2 5 of clone yb40_1 deposited under accession number ATCC 98631. In further preferred embodiments, the present invention provides a polynudeotide encoding a protein comprising a fragment of the amino acid sequence of SEQ ID N020 having biological activity, the fragment preferably comprising eight (more preferably twenty, most preferably thirty) consecutive amino acids of SEQ ID N020, or a polynucleotide encoding 3 0 a protein comprising a fragment of the amino acid sequence of SEQ ID N0:20 having biological activity, the fragment comprising ~ ammo acid sequence from amino acid 63 to amino acid 72 of SEQ ID N0:20.
Other embodiments provide the gene corresponding to the cDNA sequence of SEQ
ro NO:19.

WO 99/36512 , PCTNS99100550 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 yb40_l deposited under accession number ATCC 98631;
(ii) hybridizing said probes) to human genomic DNA in conditions at least as stringent as 4X SSC at 65 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 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 clone yb40_1 deposited under accession number ATCC 98631;
(ii) hybridizing said primers) to human genomic DNA in 2 5 conditions at least as stringent as 4X SSC at 65 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 N0:19, and 3 0 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 NO:I9. Also preferably the polynucleotide isolated according to the above process comprises a nucleotide sequence corresponding to the cDNA sequence of SEQ ID N0:19 from nucleotide 176 to nucleotide 583, and extending contiguously from a nucleotide sequence corresponding to the 5' end of said sequence of SEQ ID N0:19 from nucleotide 176 to nucleotide 583, to a nucleotide sequence corresponding to the 3' end of said sequence of SEQ ID N0:19 from nucleotide 176 to nucleotide 583.
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) fragments of the amino acid sequence of SEQ ID N0:20, each fragment comprising eight consecutive amino acids of SEQ ID N0:20; and (c) the amino acid sequence encoded by the cDNA insert of clone yb40_1 deposited under accession number ATCC 98631;
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 acid sequence of SEQ ID N0:20 having biological activity, the fragment preferably comprising eight (more preferably twenty, most preferably thirty) consecutive 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 2 0 sequence from amino acid 63 to amino acid 72 of SEQ ID N0:20.
In certain preferred embodiments, the polynucleotide is operably linked to an 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 2 5 modified expression of the genes) corresponding to the polynucleotide sequences disclosed herein.
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 3 0 (b) purifying the protein from the culture.
The protein produced according to such methods is also provided by the present 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 carrier.
BRIEF DESCRIPTION OF THE DRAWI1~TGS
Figures lA and 1B are schematic representations of the pED6 and pNOTs vectors, respectively, used for deposit of clones disclosed herein.
DETAILED DESCRIPTION
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 2 0 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 determined to be the reading frame best identifiable with sequence information available at the time of filing.
2 5 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 acid sequence. "Secreted" proteins include without limitation 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 3 0 which are transported across the membrane of the endoplasrnic reticulum.
Clone "bd577 1"
A polynucleotide of the present invention has been identified as clone "bd577 1".
bd5~ 1 was isolated from a human fetal kidney 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. bd577 1 is a full-length clone, including the entire coding sequence of a secreted protein (also referred to herein as "bd577_1 protein").
The nucleotide sequence of bd577_1 as presently determined is reported in SEQ
ID N0:1, and includes a poly(A) tail. What applicants presently believe to be the proper reading frame and the predicted amino acid sequence of the bd577 1 protein corresponding to the foregoing nucleotide sequence is reported in SEQ ID N0:2.
Amino acids 42 to 54 of SEQ ID N0:2 are a predicted leader/signal sequence, with the predicted mature amino acid sequence beginning at amino acid 55. 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 bd577 1 protein.
Another possible reading frame and predicted amino acid sequence encoded by base pairs 23 to 412 of bd577 1 SEQ ID NO:1 is reported in SEQ ID N0:30; the amino acid sequence of SEQ ID N0:30 has a possible signal sequence from amino acids 57 to 69, with the predicted mature amino acid sequence beginning at amino acid 70. The open reading frames corresponding to SEQ ID N0:30 and SEQ ID N0:2 could be joined if a frameshift 2 0 were introduced into the nucleotide sequence of SEQ ID NO:1.
The EcoRI/NotI restriction fragment obtainable from the deposit containing clone bd577_1 should be approximately 1800 bp.
The nucleotide sequence disclosed herein for bd577 1 was searched against the GenBank and GeneSeq nucleotide sequence databases using BLASTN/BLASTX and 2 5 FASTA search protocols. bd577 1 demonstrated at least some similarity with sequences identified as AA306618 (EST177563 Jurkat T-cells VI Homo sapiens cDNA 5' end) and 820055 (yg39b06.r1 Homo Sapiens cDNA clone 34805 5'). Based upon sequence similarity, bd577_1 proteins and each similar protein or peptide may share at least some activity.
The TopPredII computer program predicts two potential transmembrane domains within 3 0 the bd577 1 protein sequence centered around amino acids 42 and 230 of SEQ
ID N0:2.
bd577_1 protein was expressed in a COS cell expression system, and an expressed protein band of approximately 56 kDa was detected in membrane fractions using SDS
polyacrylamide gel electrophoresis.

S'.lone "bv280 3"
A polynucleotide of the present invention has been identified as clone '"bv280 3".
bv280_3 was isolated from a human adult brain cDNA library using methods which are selective for cDNAs encoding secreted proteins (see ~J.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. bv280 3 is a full-length clone, including the entire coding sequence of a secreted protein (also referred to herein as "bv280 3 protein').
The nucleotide sequence of bv280_3 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 bv280 3 protein corresponding to the foregoing nucleotide sequence is reported in SEQ ID N0:4.
Amino acids 10 to 22 of SEQ ID N0:4 are a predicted leader/signal sequence, with the predicted mature amino acid sequence beginning at amino acid 23. 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 bv280_3 protein.
The EcoRI/NotI restriction fragment obtainable from the deposit containing clone bv280 3 should be approximately 1900 bp.
2 0 The nucleotide sequence disclosed herein for bv280 3 was searched against the GenBank and GeneSeq nucleotide sequence databases using BLASTN/BLASTX and FASTA search protocols. bv280 3 demonstrated at least some similarity with sequences identified as AA095665 (15468.seq.F Fetal heart, Lambda ZAP Express Homo sapiens cDNA 5'), AA577430 (nm96g10.s1 NCI_CGAP_Co9 Homo Sapiens cDNA clone 2 5 1MAGE:1076130 similar to TR:G945383 6945383 CARBOXYPEPTIDASE), F06654 (H.
Sapiens partial cDNA sequence; clone c-1ga12), F08501 (H. sapiens partial cDNA), and H10119 (ym03f03.r1 Homo sapiens cDNA clone 46734 5' similar to SP:A41612 VITELLOGENIC CARBOXYPEPTZDASE). The predicted amino acid sequence disclosed herein for bv2~ 3 was searched against the GenPept and GeneSeq amino acid sequence 3 0 databases using the BLASTX search protocol. The predicted bv280_3 protein demonstrated at least some similarity to sequences identified as L46594 (carboxypeptidase [Aedes aegypti]) and 896737 (A. niger Bo-1 carboxypeptidase Y). Based upon sequence similarity, bv280_3 proteins and each similar protein or peptide may share at least some activity. The bv280_3 protein also has a serine carboxipeptidase active site motif (residues 195-212). This motif is highly specific to serine carboxypeptidases and is not found in any other type of protein in the Swiss-Prot database. The bv280 3 protein also has one copy of the crystalline beta and gamma 'Greek key' motif signature. The TopPredII
computer program predicts another potential transmembrane domain within the bv280_3 protein sequence centered around amino acid 110 of SEQ ID N0:4.
bv280 3 protein was expressed in a COS cell expression system, and an expressed protein band of approximately 61 kDa was detected in conditioned medium fractions using SDS polyacrylamide gel electrophoresis.
Clone "co315 3"
A polynucleotide of the present invention has been identified as clone "co315 3".
co315_3 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. co315 3 is a full-length clone, including the entire coding sequence of a secreted protein (also referred to herein as "co315 3 protein").
The nucleotide sequence of co315 3 as presently determined is reported in SEQ
ID N0:5, 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 co315 3 protein corresponding to the foregoing nucleotide sequence is reported in SEQ ID N0:6.
Amino acids 51 to 63 of SEQ ID N0:6 are a predicted leader/signal sequence, with the predicted mature amino acid sequence beginning at amino acid 64. 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 co315 3 protein.
The EcoRI/Notl restriction fragment obtainable from the deposit containing clone co315_3 should be approximately 710 bp.
The nucleotide sequence disclosed herein for co315 3 was searched against the 3 0 GenBank and GeneSeq nucleotide sequence databases using BLASTN/BLASTX and FASTA search protocols. co315_3 demonstrated at least some similarity with sequences identified as AA031371 (zk15e11.s1 Soares pregnant uterus NbHPU Homo Sapiens cDNA
clone 470636 3'), AA026051 (ze86a07.s1 Soares fetal heart NbHHI9W Homo Sapiens), AA393961 (zt78b10.r1 Soares testis NHT Homo sapiens cDNA clone 728443 5'), (aa29c06.s1 NCI_CGAP GCBl Homo sapiens cDNA clone IMAGE:814666 3'), H46323 (yo15c05.r1 Homo sapiens cDNA clone 177992 5'), N23329 (yx78h09.s1 Homo sapiens cDNA clone 267905 3'), and 843942 (yg22f02.s1 Homo sapiens cDNA clone 33080 3' similar to gb:M14648 VTTRONECTIN RECEPTOR ALPHA SUBUNIT PRECURSOR
(HUMAN)). Based upon sequence similarity, co315_3 proteins and each similar protein or peptide may share at least some activity.
lone "ij226 6"
A polynucleotide of the present invention has been identified as clone "ij226_6".
ij226_6 was isolated from a human adult blood (peripheral blood mononuclear cells treated with granulocyte-colony stimulating factor in vivo) 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. ij226_6 is a full-length clone, including the entire coding sequence of a secreted protein (also referred to herein as "ij226_6 protein'.').
The nucleotide sequence of ij226_6 as presently determined is reported in SEQ
ID
N0:7, and includes a poly(A) tail. What applicants presently believe to be the proper reading frame and the predicted amino acid sequence of the ij226_6 protein corresponding 2 0 to the foregoing nucleotide sequence is reported in SEQ ID NO:B.
The EcoRI/NotI restriction fragment obtainable from the deposit containing clone ij226_6 should be approximately 2300 bp.
The nucleotide sequence disclosed herein for ij226_6 was searched against the GenBank and GeneSeq nucleotide sequence databases using BLASTN/BLASTX and FASTA search protocols. ij226_6 demonstrated at least some similarity with sequences identified as AE000658 (Homo sapiens T-cell receptor alpha delta locus from bases 1 to 250529 (section 1 of 5) of the Complete Nucleotide Sequence), AF004231 (Homo Sapiens monocyte/macrophage Ig-related receptor MIR-10 (Milt cl-10) mRNA, complete cds), 635352 (STS h14a108 5), H54023 (yq88hOl.s1 Homo sapiens cDNA), H54181 (yq88hOl.r1 3 0 Homo sapiens cDNA clone 202897 5'), T18551 (Human polycystic kidney disease normal PKD1 gene), and 282206 (Human DNA sequence *** SEQUENCING IN PROGRESS ***
from clone 370M22; HTGS phase 1). The predicted amino acid sequence disclosed herein for ij226_6 was searched against the GenPept and GeneSeq amino acid sequence databases using the BLASTX search protocol. The predicted ij226_6 protein demonstrated at least some similarity to sequences identified as M22334 (unknown protein [Homo sapiens]):
Based upon sequence similarity, ij226 6 proteins and each similar protein or peptide may share at least some activity. The TopPredII computer program predicts two 'potential transmembrane domains within the ij226_6 protein sequence centered around amino acids 37 and 62 of SEQ ID N0:8. The nucleotide sequence of ij226_6 indicates that it may contain one or more of the following repetitive elements: Ll, Alu, SVA.
Clone"nf443 1"
A polynucleotide of the present invention has been identified as clone "nf443_1".
nf443_1 was isolated from a human adult brain (substantia nigra) 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.
nf443_1 is a full-length clone, including the entire coding sequence of a secreted protein (also referred to herein as "nf443_1 protein").
The nucleotide sequence of nf443_1 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 nf443_1 protein corresponding to the foregoing nucleotide sequence is reported in SEQ ID
NO:10. Amino 2 0 acids 21 to 43 of SEQ ID N0:10 are a possible leader/signal sequence, with the predicted mature amino acid sequence beginning at amino acid 44. Due to the hydrophobic nature of this possible leader/signal sequence, it is likely to act as a transmembrane domain should the leader/signal sequence not be separated from the remainder of the nf443_1 protein.
2 5 The EcoRI/NotI restriction fragment obtainable from the deposit containing clone nf443_1 should be approximately 3800 bp.
The nucleotide sequence disclosed herein for nf443_1 was searched against the GenBank and GeneSeq nucleotide sequence databases using BLASTN/BLASTX and FASTA search protocols. nf443_1 demonstrated at least some similarity with sequences 3 0 identified as AA417092 (zu07a12.s1 Soares testis NHT Homo Sapiens cDNA
clone 731134 3'), AA421511 (zu07aI2.r1 Soares testis NHT Homo Sapiens cDNA clone 731134 5'), T23707 (Human gene signature HUMGS05583), and U61233 (Bos taurus tubulin-folding cofactor D mRNA, complete cds). The predicted amino acid sequence disclosed herein for nf443_1 was searched against the GenPept and GeneSeq amino acid sequence databases using the BLASTX search protocol. The predicted nf443_1 protein demonstrated at least some similarity to sequences identified as U61233 (cofactor D [Bos taurusj). Based upon sequence similarity, nf443_1 proteins and each similar protein or peptide may share at least some activity. The nucleotide sequence of nf443_1 indicates that it may contain an Alu repetitive element.
nf443_1 protein was expressed in a COS cell expression system, and an expressed protein band of approximately 10 kDa was detected in conditioned medium fractions using SDS polyacrylamide gel electrophoresis.
Clone "nt429 1"
A polynucleotide of the present invention has been identified as clone "nt429_1".
nt429_1 was isolated from a human adult brain (corpus callosum) cDNA library using methods which are selective for cDNAs encoding secreted proteins (see U.S.
Pat. No.
5,53b,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.
nt429_1 is a full-length done, including the entire coding sequence of a secreted protein (also referred to herein as "nt429_1 protein').
The nucleotide sequence of nt429_1 as presently determined is reported in SEQ
ID
NO:11, and includes a poly(A) tail. What applicants presently believe to be the proper 2 0 reading frame and the predicted amino acld sequence of the nt429_1 protein corresponding to the foregoing nucleotide sequence is reported in SEQ ID
N0:12.
Another possible reading frame and predicted amino acid sequence, encoded by base pairs 399 to 731 of nt429_1 SEQ ID NO:11, is reported in SEQ ID N0:31; the amino acid sequence of SEQ ID N0:31 is hydrophobic in nature near its carboxyl terminus.
The 2 5 overlapping open reading frames corresponding to SEQ ID N0:12 and SEQ ID
N0:31 could be joined if a frameshift were introduced into the nucleotide sequence of SEQ ID
N0:11.
The EcoRI/NotI restriction fragment obtainable from the deposit containing done nt429_1 should be approximately 1800 bp.
3 0 The nucleotide sequence disclosed herein for nt429_1 was searched against the GenBank and GeneSeq nucleotide sequence databases using BLASTN/BLASTX and FASTA search protocols. No significant hits were found in the database. The nucleotide sequence of nt429_1 indicates that it may contain one or more of the following repetitive elements: Alu, SVA, A.

QlQne " , en 503 1_' A polynucleotide of the present invention has been identified as clone "pe503_1".
pe503_1 was isolated from a human adult blood (chronic myelogenous leukemia IC5) 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. pe503_1 is a full-length clone, including the entire coding sequence of a secreted protein (also referred to herein as "pe503_1 protein").
The nucleotide sequence of pe503_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 reading frame and the predicted amino acid sequence of the pe503_1 protein corresponding to the foregoing nucleotide sequence is reported in SEQ ID
N0:14. Amino ands 79 to 91 of SEQ ID N0:14 are a predicted leader/signal sequence, with the predicted mature amino acid sequence beginning at amino acid 92. 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 pe503_1 protein.
The EcoRI/NotI restriction fragment obtainable from the deposit containing clone pe503_1 should be approximately 1300 bp.
2 0 The nucleotide sequence disclosed herein for pe503_1 was searched against the GenBank and GeneSeq nucleotide sequence databases using BLASTN/BLASTX and FASTA search protocols. pe503_i demonstrated at least some similarity with sequences identified as AA298572 (EST114204 HSC172 cells I Homo Sapiens cDNA 5' end), AA595242 (no33a12.s1 NCI_CGAP_Pr23 Homo sapiens cDNA clone IMAGE:1102462), 2 5 H60941 (yr14g06.r1 Homo sapiens cDNA clone 205306 5'), H75686 (yr77g08.r1 Homo sapiens cDNA clone 211358 5'), and 861206 (yh06d11.r1 Homo sapiens cDNA clone 5'). Based upon sequence similarity, pe503_1 proteins and each similar protein or peptide may share at least some activity. The TopPredII computer program predicts four potential transmembrane domains within the pe503_1 protein sequence centered around amino 3 0 acids 50, 84,107, and 148 of SEQ ID N0:14, respectively.
pe503_1 protein was expressed in a COS cell expression system, and an expressed protein band of approximately 19 kDa was detected in membrane fractions using SDS
polyacrylamide gel electrophoresis.

Close "pe834 6"
A polynucleotide of the present invention has been identified as clone "pe834_6".
pe834_6 was isolated from a human adult blood (chronic myelogenous leukemia K5) 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. pe834_6 is a full-length clone, including the entire coding sequence of a secreted protein (also referred to herein as "pe834_6 protein").
The nucleotide sequence of pe834_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 pe834_6 protein corresponding to the foregoing nucleotide sequence is reported in SEQ ID
N0:16.
Another possible reading frame and predicted amino acid sequence, encoded by base pairs 414 to 725 of pe834_6 SEQ ID N0:15, is reported in SEQ ID N0:32; the amino acid sequence of SEQ ID N0:32 is hydrophobic in nature near its carboxyl terminus.
The overlapping open reading frames corresponding to SEQ ID N0:16 and SEQ ID N0:32 could be joined if a frameshift were introduced into the nucleotide sequence of SEQ ID
N0:15.
The EcoRI/NotI restriction fragment obtainable from the deposit containing clone 2 0 pe834_6 should be approximately 1300 bp.
The nucleotide sequence disclosed herein for pe834_6 was searched against the GenBank and GeneSeq nucleotide sequence databases using BLASTN/BLASTX and FASTA search protocols. pe834_6 demonstrated at least some similarity with sequences identified as AA054341 (z168f04.s1 Stratagene colon (#937204) Homo Sapiens cDNA clone 2 5 5097913'), N21462 (yx57c10.s1 Homo sapiens cDNA clone 265842 3'), N34010 (yx75g07.r1 Homo sapiens cDNA clone 267612 5'), and T90232 (ye15c09.r1 Homo Sapiens cDNA
clone 117808 5'). Based upon sequence similarity, pe834_6 proteins and each similar protein or peptide may share at least some activity.
pe834 6 protein was expressed in a COS cell expression system, and an expressed 3 0 protein band of approximately 17 kDa was detected in membrane fractions using SDS
polyacrylamide gel electrophoresis.

Clone "yal0 1"
A polynucleotide of the present invention has been identified as clone "yal0_1".
yal0_1 was isolated from a human adult testes 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. yal0_1 is a full-length clone, including the entire coding sequence of a secreted protein (also referred to herein as "yal0_1 protein").
The nucleotide sequence of yal0_1 as presently determined is reported in SEQ
ID
N0:17, and includes a poly(A) tail. What applicants presently believe to be the proper reading frame and the predicted amino acid sequence of the yal0_1 protein corresponding to the foregoing nucleotide sequence is reported in SEQ ID N0:18. Amino acids 6 to 18 of SEQ ID N0:18 are a predicted leader/signal sequence, with the predicted mature amino acid sequence beginning at amino acid 19. 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 yal0_1 protein.
The EcoRI/NotI restriction fragment obtainable from the deposit containing clone yal0_1 should be approximately 800 bp.
The nucleotide sequence disclosed herein for yal0_1 was searched against the GenBank and GeneSeq nucleotide sequence databases using BLASTN/BLASTX and 2 0 FASTA search protocols. No clearly significant hits were found in these databases.
BLASTX analysis of the yal0_1 protein sequence revealed some amino acid sequence similarity to cystatins (cysteine protease inhibitors) of various species.
Based upon this sequence similarity, yal0_1 proteins and each similar protein or peptide may share at least some activity.
Clone "3rb40 1"
A polynucleotide of the present invention has been identified as clone "yb40_1".
yb40_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 3 0 amino acid sequence of the encoded protein. yb40_1 is a full-length clone, including the entire coding sequence of a secreted protein (also referred to herein as "yb40_1 protein") The nucleotide sequence of yb40_1 as presently determined is reported in SEQ
ID
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 yb40_1 protein corresponding to the foregoing nucleotide sequence is reported in SEQ ID
N0:20. Amino acids 29 to 41 of SEQ ID N0:20 are a possible leader/signal sequence, with the predicted mature amino acid sequence begirming at amino acid 42. Due to the hydrophobic nature of this possible leader/signal sequence, it could act as a transmembrane domain should it not be separated from the remainder of the yb40_1 protein.
The EcoRI/NotI restriction fragment obtainable from the deposit containing clone yb40_1 should be approximately 1700 bp.
The nucleotide sequence disclosed herein for yb40_1 was searched against the GenBank and GeneSeq nucleotide sequence databases using BLASTN/BLASTX and FASTA search protocols. yb40_1 demonstrated at least some similarity with sequences identified as AA595189 (no32f03.s1 NCI_CGAP_Pr23 Homo Sapiens cDNA clone 1MAGE:I102397), 874575 (yi58d04.r1 Homo sapiens cDNA clone 1434315'), and (Human gene signature HUMGS08001). Based upon sequence similarity; yb40_1 proteins and each similar protein or peptide may share at least some activity.
Deposit of Clones Clones bd577_1, bv280_3, co315 3, ij226_6, nf443_1, nt429_l, pe503_l, pe834_6, yal0_1, and yb40_1 were deposited on January 13,1998 with the American Type Culture Collection (10801 University Boulevard, Manassas, Virginia 20110-2209 U.S.A.}
as an 2 0 original deposit under the Budapest Treaty and were given the accession number ATCC
98631, 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.
2 5 Each clone has been transfected into separate bacterial cells {E. coh~ 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 3 0 ("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 was derived from pMT2 (Kaufman et al.,1989, MoI. 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 some 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 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:
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.
Clone Probe uence bd577-1 SEQ ID N0:21 bv280 3 SEQ ID N0:22 co315 3 SEQ ID N0:23 ij226_6 SEQ ID N0:24 nf443_1 SEQ ID N0:25 2 0 nt429_1 SEQ ID N0:26 pe503_1 SEQ ID N0:27 pe834_6 SEQ ID N0:28 yb40_1 SEQ ID N0:29 2 5 1n 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)}.
3 0 The design of the oligonucleotide probe should preferably follow these parameters:
(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 Tm 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 'ZP 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 probe should be approximately 4e+6 dpm/pmole.
The bacterial culture containing the pool of full-length clones should preferably be thawed and 100 l,xl of the stock used to inoculate a sterile culture flask containing 25 ml of sterile L-broth containing ampicillin at 100 llg/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 llg/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 2 0 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 llg/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 2 5 a concentration greater than or equal to le+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 3 0 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.

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 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. ~4, 9245-9253 (1992), both of which are incorporated herein by reference. Such fragments may be fused to earner molecules such as immunoglobulins for many purposes, including increasing the valency of protein binding 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 2 0 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 2 5 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 3 0 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 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 polynucleotide by identifying significantly similar nucleotide sequences in public databases, such as expressed sequence tags {SSTs), 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.
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 "UniGene clusters"
so identified will already have been mapped to particular chromosomal sites.
Organisms that have enhanced, reduced, or modified expression of the gene{s) corresponding to the polynucleotide sequences disclosed herein are provided.
The desired change in gene expression can be achieved through the use of antisense 2 0 polynucleotides or ribozymes that bind and/or cleave the mRNA transcribed from the gene (Albert and Morris,1994, Trends Pharmacol. Sci.15(7): 250-254; Iravarosky et al.,1997, Biochem. Mol. Med. 62(1):11-22; and Hampel,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 2 5 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 B1, incorporated by reference herein).
3 0 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.
Acad. Sci. LISA 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 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).
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 2 0 location of the center of the transmsmbrane domain, with at Least ten transmembrane amino acids on each side of the reported central residues}.
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 2 5 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 3 0 (more preferably 20 or more, most preferably 30 or more) contiguous amino acids 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 I.4, which in turn is based on the public domain NCBI-BLAST version 1.4 (Altschul and Gish, 199b, 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: 26b-272; Karlin and Altschul, 1993, Applications and statistics for multiple high-scoring segments in molecular sequences, Proc. Natl. Acad. Sci. USA 90: 5873-587?; 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 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 2 0 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 2 5 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 BLOSUM62, but other amino acid comparison matrices such as PAM can be utilized.
Species homologues of the disclosed polynucleotides and proteins are also 3 0 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 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 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 and 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 species such as, for example, Pan troglodytes, Gorilla gorilla, Pongo pygrnaeus, Hylobates concolor, Macaca mulatta, Papio papio, Papio hamadryas, Cercopithecus aethiops, Cebus capucinus, Aotus trivirgatus, Sanguinus Oedipus, Microcebus murinus, Mus musculus, Rattus norvegicus, Cricetulus griseus, Fells cntus, Mustela vison, Cams familiaris, 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 Seu~nez, 1988, Ann. Rev. Genet. 22: 323-351; O'Brien et aL, 1993, Nature 2 0 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 2 5 which also encode proteins which are identical or have significantly similar sequences to those encoded by the disclosed polynucleotides. 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 3 0 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.
SO

The invention also includes polynucleotides with sequences complementary to those of the polynudeotides disclosed herein.
The present invention also includes polynueleotides 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 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.

StringencyPolynucleotideHybridHybridization TemperatureWash ConditionHybrid Lengthand Temperature (bp)$ Buffers and Buffers A DNA:DNA 2 50 65C; lxSSC-or- 65C; 0.3xSSC
42C; lxSSC, 50% formamide B DNA:DNA <50 TB*; lxSSC TB*; lxSSC

C DNA:RNA Z 50 67C; lxSSC -or- 67C; 0.3xSSC
45C; lxSSC, 50% formamide D DNA:RNA <50 Tp*; lxSSC TD*; lxSSC

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

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

I DNA:RNA 2 50 67C; 4xSSC -or- 67C; lxSSC
45C; 4xSSC, 50% formamide J DNA:RNA <50 T~*; 4xSSC TJ*; 4xSSC

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

M DNA:DNA 2 50 50C; 4xSSC -or- 50C; 2xSSC
40C; 6xSSC, 50~ formamide N DNA:DNA <50 TN*; 6xSSC TN*; 6x~C

O DNA:RNA 2 50 55C; 4xSSC -or- 55C; 2xSSC
42C; 6xSSC, 50% formamide P DNA:RNA c50 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

#: The hybrid length is that anticipated for the hybridized regions) of the hybridizing polynucleotides. When hybridizing a polynudeotide to a target polynudeotide of unknown sequence, the hybrid length is assumed to be that of the hybridizing polynudeotide. 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 rnmplementarity.
f: SSPE (lxSSPE is 0.15M NaCI, lOmM NaH2P0*, and 1.25mM EDTA, pH 7.4) can be substituted for SSC
(lxSSC is 0.15M NaCI and l5mM 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 T," is determined according to the following equations. For hybrids less than 18 base pairs in length, T,"(°C) = 2(# of A + T bases) + 4(# of G +
C bases). For hybrids between 18 and 49 base pairs in length, T°,(°C) = 81.5 + 16.6(log,o[Na'j) + 0.41 (%G+C) (600/N), where N is the number of bases in the hybrid, and jNa'j is the concentration of sodium ions in the 3 5 hybridization buffer ([Na'j 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, lnc., 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 palynucleotide 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 pM1'2 or pED expression vectors disclosed in Kaufman et al., Nucleic Acids Res. ~9, 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 5 537-566 (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 vitro 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 Agricultural Experiment Station Bulletin No 1555~1987~, 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, Nn 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 (1ZP-HPLC) steps employing hydrophobic 1ZP-HPLC media, e.g., silica gel having pendant methyl or other aliphatic groups, can be employed to further purify the protein. Sorne 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 SS

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 Uses 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 2 0 (when labeled) to identify chromosomes or to map related gene positions;
to compare 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 2 5 and making oligomers for attachment to a "gene chip" or other support, including for 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 3 0 polynucleotide can also be used in interaction trap assays (such as, for example, those described in Gyuris et al.,1993, CeII 75: 791-803 and in Rossi et al.,1997, Proc. Natl. Acad.
Sci. LISA 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 agonists 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. Maniatis 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.
C;~rtokine and Cell Proliferation/Differentiation Activity r 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 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, T20, B9, B9/11, BaF3, MC9/G, M+ (preB M+), 2E8, RBS, DA1,123, T1165, HT2, CTLL2, TF-1, Mo7e 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. Immunol.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 2 0 cells or thymocytes include, without limitation, those described in:
Polyclonal T cell stimulation, Kruisbeek, A.M. and Shevach, E.M. In Current Protocols in Immunology. J.E.e.a.
Coligan eds. VoI 1 pp. 3.12.1-3.12.14, John Wiley and Sons, Toronto. 1994; and Measurem~t 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.
2 5 Assays for proliferation and differentiation of hematopoietic and lymphopoietic cells include, without limitation, those described in: Measurement of Human and Murine Interleukin 2 and Interleukin 4, Bottomly, 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 3 0 336:690-692, 1988; Greenberger et al., Proc. Natl. Acad. Sci. U.S.A.
80:2931-2938, 1983;
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 lnterleukin 11- Bennett, 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 Interleukin 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. 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; 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.
11:405-4I1, 1981; Takai et al., J. Immunol. 137:3494-3500, 1986; Takai et al., J. Immunol.
140:508-512, 1988.
Immune Stimulating or u~nr-,, ecs,'_n~ Art;vi 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 2 0 deficiencies and disorders (including severe combined immunodeficiency (SCID)), e.g., 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 autoirnrnune disorders. More specifically, infectious 2 5 diseases causes by viral, bacterial, fungal or other infection may be treatable using a protein of the present invention, including infections by HIV, hepatitis viruses, herpesviruses, mycobacteria, Leishmania spp., malaria spp. and various fungal infections such as candidiasis. C~f 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 3 0 treatment of cancer.
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 aMivated 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 2 0 limitation B lymphocyte antigen functions (such as , for example, B7)), e.g., preventing 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 2 5 through its recognition as foreign by T cells, followed by an immune reaction that destroys 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., B7-3 0 1, B7-3) or blocking antibody), prior to transplantation can lead to the binding of the 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 bloclting 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 2 0 activation of autoreactive T cells may reduce or eliminate disease symptoms.
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 2 5 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/~pr/lpr mice or NZB hybrid mice, 3 0 marine autoimmune collagen arthritis, diabetes mellitus in NOD mice and BB
rats, and 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, 2 0 carcinoma) transfected with a nucleic acid encoding at least one peptide of the present 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 2 5 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. Altenlatively, 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
3 0 lymphocyte antigens) on the surface of the tumor cell provides the necessary 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 (i2 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.
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); Herrmann et al., Proc. Natl.
Acad. Sci.
2 0 USA 78:2488-2492,1981; Herrmann et al., J. Immunol.128:1968-1974,1982;
Handa et al., J. Immunol. 135:1564-1572,1985; Takai et al., J. Immunol.137:3494-3500,1986;
Takai et al., ]. Immunol.140:508-512, 1988; Herrmann et al., Proc. Natl. Acad. Sci. USA
78:2488-2492, 1981; Herrmann et al., J. Immunol. 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.
2 5 Virology 61:1992-1998; Takai et al., J. Immunol. 140:508-512, 1988;
Bertagnolli et al., Cellular Immunology 133:327-341,1991; Brown et al., J. Immunol.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 Thl /Th2 profiles) include, without limitation, those described 3 0 in: Maliszewski, J. Immunol.144:3028-3033,1990; and Assays for B cell function: In vitro 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 Th1 and CTL responses) 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; 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, 2 0 Journal of Immunology 145:4037-4045, 1990; Zamai et al., Cytometry 14:891-897, 1993;
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 2 5 85:2770-2778,1995; Toki et al., Proc. Nat. Acad Sci. USA 88:7548-7551, 1991.
Hematonoiesis Re~ulatin~g Actin 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 3 0 marginal biological activity in support of colony forming cells or of factor-dependent cell 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 WO 99/36512 ' PCTNS99/00550 precursors and/or enythroid 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 comparhnent 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.
2 0 , Assays for embryonic stem cell differentiation (which will identify, among others, 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.
2 5 Assays for stem cell survival and differentiation (which will identify, among 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 3 0 hematopoietic colony forming cells with high proliferative potential, McNiece, LK. and Briddell, R.A. In Culture of Hematopoietic Cells. R.I. Freshney, et aI. 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 aI. 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 HematopoieHc Cells. R.I. Freshney, et aI. eds. Vol pp.
163-179, Wiley-Liss, Inc., New York, NY.1994; Long term culture initiating cell assay, Sutherland, H.J. In Culture of Hemutopoietic Cells. R.I. Freshney, et al. eds. Vol pp.139-162, Wiley-Liss;
Inc., New York, NY.1994.
Tissue Growth Activi.tv 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.
2 0 A protein of this invention may also be used in the treatment of periodontal 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 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 3 0 invention, which induces tendon/ligament-like tissue or other tissue formation in 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 WO 99136512 PCT/US99l00550 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 2 0 localized neuropathies, and central nervous system diseases, such as Alzheimer's, Parkinson's disease, Huntington'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 2 5 from chemotherapy or other medical therapies may also be treatable using a protein of the 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.
3 0 It is expected that a protein of the present invention may also exhibit activity for 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 angiogeruc 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, Inc., Chicago, as modified by Eaglstein and Mertz, J.
Invest.
Dermatol 71:382-84 (1978).
2 0 Activin/Inhibin Activit~r 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 ~3 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.
Chemotactic/Chemokinetic Activity A protein of the present invention may have chemotactic or chemokinetic activity (e.g., act as a chemokine) for 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. lmmunol. 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 Thrombo~rtic Activity 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 hemophiliac) or to enhance coagulation and other hemostadc 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. Olin. 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.
2 0 Receptor/Ligand 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 A
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 l:L-1. Proteins of the invention may also be useful to treat anaphylaxis and hypersensitivity to an antigenic substance or material.
Cadhezin/Tumor Invasion Su,~nressnr Activity 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 I O 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 Activity 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 carrier.
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 tk~e 2 0 effectiveness of the biological activity of the active ingredient{s). The characteristics of the 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, ILr.7, IIr8, IL-9, IL-10, ILrll, IL-12, IL-13,1L-14, IL-15, IFN, TNFO, TNFl, TNF2, G-CSF, Meg-CSF, thrombopoietin, stem 2 5 cell factor, and erythropoietin. The pharmaceutical composition may further contain other 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 3 0 in formulations of the particular cytokine, lymphokine, other hematopoietic factor, 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 2 0 pharmaceutically acceptable earners, with amphipathic agents such as lipids which exist in aggregated form as micelles, insoluble monolayers, liquid crystals, or lamellar layers 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 2 5 art, as disclosed, for example, in U.S. Patent No. 4,235,871; U.S. Patent No. 4,501,728; U.S.
Patent No. 4,837,028; and U.S. Patent No. 4,737,323, all of which are incorporated herein 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 3 0 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, 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 cytokines, lymphokines or other hematopoietic factors. When co-administered with one or more cytokines, lymphokines or other hematopoietic factors, protein of the present invention may be administered either simultaneously with the cytokine(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 cytokine(s), lymphokine(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.
2 0 Intravenous administration to the patient is preferred.
When a therapeutically effective amount of protein of the present invention is 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 2 5 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.
When administered in liquid form, a liquid earner 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 3 0 physiological saline solution, dextrose or other saccharide solution, or glycols such as ethylene glycol, propylene glycol or polyethylene glycol. When administered in liquid form, the pharmaceutical composition rnntains 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 Ringer'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 2 0 increased further. It is contemplated that the various pharmaceutical compositions used to practice the method of the present invention should contain about 0.01 llg to about 100 mg (preferably about O.lng to about 10 mg, more preferably about 0.1 pg 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 2 5 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 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 3 0 therapy using the pharmaceutical composition of the present invention.
Protein of the invention may also be used to immunize animals to obtain polyclonal and monoclonal antibodies which specifically react with the protein. Such antibodies may be obtained using either the entire protein or fragments thereof as an immunogen. The peptide immunogens additionally may contain a cysteine residue at the carboxyl terminus, and are conjugated to a hapten such as keyhole limpet hemocyarun (KL~. Methods for synthesizing such peptides are known in the art, for example, as in R.P. Merrifield, J. Amer.Chem.Soc. ~S, 2149-2154 (1963); J.L. Krstenansky, et aL, FEBS Left.
211. 10 (1987). Monoclonal antibodies binding to the protein of the invention may be useful 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 2 0 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 2 5 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 applications.
The choice of matrix material is based on biocompatibility, biodegradability, mechanical properties, cosmetic appearance and interface properties. The particular 3 0 application of the compositions will define the appropriate formulation.
Potential matrices for the compositions may be biodegradable and chemically defined calcium 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 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 2 0 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.
2 5 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 question. These agents include various growth factors such as epidermal growth factor (EGF), platelet den~ived growth factor (PDGF}, transforming growth factors (TGF-a and TGF-~3), and insulin-like growth factor (IGF).
3 0 The therapeutic compositions are also presently valuable for veterinary applications. Particularly domestic animals and thoroughbred horses, in addition to 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 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.
2 0 Patent and literature references cited herein are incorporated by reference as if fully set forth.

SEQUENCE LISTING
<110> 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.
Spaulding, Vikki along, Gordon G.
Clark, Hilary Fechtel, Rim Genetics Institute, Inc.
<120> SECRETED PROTEINS AND POLYNUCLEOTIDES ENCODING THEM
<130> GI 6061A
<140>
<141>
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<213> Homo sapiens <400> 2 Met Glu Trp Trp Trp Asp Cys Leu Glu Met Leu Arg Glu Asn Thr Leu Val Thr Leu Ala Asn Ile Ser Gly Gln Leu Asp Leu Ser Pro Tyr Pro Glu Ser Ile Cys Leu Pro Val Leu Asp Gly Leu Leu His Trp Ala Val Cys Pro Ser Ala Glu Ala Gln Asp Pro Phe Ser Thr Leu Gly Pro Asn Ala Val Leu Ser Pro Gln Arg Leu Val Leu Glu Thr Leu Ser Lys Leu Ser Ile Gln Asp Asn Asn Val Asp Leu Ile Leu Ala Thr Pro Pro Phe 85 90 g5 Ser Arg Leu Glu Lys Leu Tyr Ser Thr Met Val Arg Phe Leu Ser Asp Arg Lys Asn Pro Val Cys Arg Glu Met Ala Val Val Leu Leu Ala Asn Leu Ala Gln Gly Asp Ser Leu Ala Ala Arg Ala Ile Ala Val Gln Lys Gly Ser Ile Gly Asn Leu Leu Gly Phe Leu Glu Asp Ser Leu Ala Ala Thr Gln Phe Gln Gln Ser Gln Ala Ser Leu Leu His Met Gln Asn Pro Pro Phe Glu Pro Thr Ser Val Asp Met Met Arg Arg Ala Ala Arg Ala Leu Leu Ala Leu Ala Lys Val Asp Glu Asn His Ser Glu Phe Thr Leu Tyr Glu Ser Arg Leu Leu Asp Ile Ser Val Ser Pro Leu Met Asn Ser Leu Val Ser Gln Val Ile Cys Asp Val Leu Phe Leu Ile Gly Gln Ser <210> 3 <211> 1684 <212> DNA
<213> Homo sapiens <400> 3 ctgcctgatt tgggaagcgc tgcaaggaca accggctggg gtccttgcgc gccgcggctc 60 agggaggagc accgactgcg ccgcaccctg agagatggtt ggtgccatgt ggaaggtgat 120 tgtttcgctg gtcctgttga tgcctggccc ctgtgatggg ctgtttcact ccctatacag 180 aagtgtttcc atgccaccta agggagactc aggacagcca ttatttctca ccccttacat 240 tgaagctggg aagatccaaa aaggaagaga attgagtttg gtcggtcctt tcccaggact 300 gaacatgaag agttatgccg gcttcctcac cgtgaataag acttacaaca gcaacctctt 360 cttctggttc ttcccagctc agatacagcc agaagatgcc ccagtagttc tctggctaca 420 gggtgggccg ggaggttcat ccatgtttgg actctttgtg gaacatgggc cttatgttgt 480 cacaagtaac atgaccttgc gtgacagaga cttcccctgg accacaacgc tctccatgct 540 ttacattgac aatccagtgg gcacaggctt cagttttact gatgataccc acggatatgc 600 agtcaatgag gacgatgtag cacgggattt atacagtgca ctaattcagt ttttccagat 660 atttcctgaa tataaaaata atgactttta tgtcactggg gagtcttatg cagggaaata 720 tgtgccagcc attgcacacc tcatccattc cctcaaccct gtgagagagg tgaagatcaa 780 cctgaacgga attgctattg gagatggata ttctgatccc gaatcaatta tagggggcta 840 tgcagaattc ctgtacctaa ttggcttgtt ggatgagaag caaaaaaagt acttccagaa 900 gcagtgccat gaatgcatag aacacatcag gaagcagaac tggtttgagg cctttgaaat 960 actggataaa ctactagatg gcgacttaac aagtgatcct tcttacttcc agaatgttac 1020 aggatgtagt aattactata actttttgcg gtgcacggaa cctgaggatc agctttacta 1080 tgtgaaattt ttgtcactcc cagaggtgag acaagccatc cacgtgggga atcagacttt 1140 taatgatgga actatagttg aaaagtactt gcgagaagat acagtacagt cagttaagcc 1200 atggttaact gaaatcatga ataattataa ggttctgatc tacaatggcc aactggacat 1260 catcgtggca gctgccctga cagagcgctc cttgatgggc atggactgga aaggatccca 1320 ggaatacaag aaggcagaaa aaaaagtttg gaagatcttt aaatctgaca gtgaagtggc 1380 tggttacatc cggcaagcgg gtgacttcca tcaggtaatt attcgaggtg gaggacatat 1440 tttaccctat gaccagcctc tgagagcttt tgacatgatt aatcgattca tttatggaaa 1500 aggatgggat ccttatgttg gataaactac cttcccaaaa gagaacatca gaggttttca 1560 ttgctgaaaa gaaaatcgta aaaacagaaa atgtcatagg aataaaaaaa ttatcttttc 1620 atatctgcaa gatttttttc atcaataaaa attatccttg raaaaaaaaa aaaaaaaaaa 1680 ease 1684 <210> 4 <211> 476 <212> PRT
<213> Homo sapiens <400> 4 Met Val Gly Ala Met Trp Lys Val Ile Val Ser Leu Val Leu Leu Met Pro Gly Pro Cys Asp Gly Leu Phe His Ser Leu Tyr Arg Ser Val Ser Met Pro Pro Lys Gly Asp Ser Gly Gln Pro Leu Phe Leu Thr Pro Tyr Ile Glu Ala Gly Lys Ile Gln Lys Gly Arg Glu Leu Ser Leu Val Gly Pro Phe Pro Gly Leu Asn Met Lys Ser Tyr Ala Gly Phe Leu Thr Val Asn Lys Thr Tyr Asn Ser Asn Leu Phe Phe Trp Phe Phe Pro Ala Gln B5 ' 90 95 Ile Gln Pro Glu Asp Ala Pro Val Val Leu Trp Leu Gln Gly Gly Pro Gly Gly Ser Ser Met Phe Gly Leu Phe Val Glu His Gly Pro Tyr Val Val Thr Ser Asn Met Thr Leu Arg Asp Axg Asp Phe Pro Trp Thr Thr Thr Leu Ser Met Leu Tyr Ile Asp Asn Pro Val Gly Thr Gly Phe Ser Phe Thr Asp Asp Thr His Gly Tyr Ala Val Asn Glu Asp Asp Val Ala Arg Asp Leu Tyr Ser Ala Leu Ile Gln Phe Phe Gln Ile Phe Pro Glu Tyr Lys Asn Asn Asp Phe Tyr Val Thr Gly Glu Ser Tyr Ala Gly Lys Tyr Val Pro Ala Ile Ala His Leu Ile His Ser Leu Asn Pro Val Arg Glu Val Lys Ile Asn Leu Asn Gly Ile Ala Ile Gly Asp Gly Tyr Ser Asp Pro Glu Ser Ile Ile Gly Gly Tyr Ala Glu Phe Leu Tyr Leu Ile Gly Leu Leu Asp Glu Lys Gln Lys Lys Tyr Phe Gln Lys Gln Cys His Glu Cys Ile Glu His Ile Arg Lys Gln Asn Trp Phe Glu Ala Phe Glu Ile Leu Asp Lys Leu Leu Asp Gly Asp Leu Thr Ser Asp Pro Ser Tyr Phe Gln Asn Val Thr Gly Cys Ser Asn Tyr Tyr Asn Phe Leu Arg Cys Thr Glu Pro Glu Asp Gln Leu Tyr Tyr Val Lys Phe Leu Ser Leu Pro Glu Val Arg Gln Ala Ile His Val Gly Asn Gln Thr Phe Asn Asp Gly Thr Ile Val Glu Lys Tyr Leu Arg Glu Asp Thr Val Gln Ser Val Lys Pro Trp Leu Thr Glu Ile Met Asn Asn Tyr Lys Val Leu Ile Tyr Asn Gly Gln Leu Asp Ile Ile Val Ala Ala Ala Leu Thr Glu Arg Ser Leu Met Gly Met Asp Trp Lys Gly Ser Gln Glu Tyr Lys Lys Ala Glu Lys Lys Val Trp Lys Ile Phe Lys Ser Asp Ser Glu Val Ala Gly Tyr Ile Arg Gln Ala Gly Asp Phe His Gln Val Ile Ile Arg Gly Gly Gly His Ile Leu Pro Tyr Asp Gln Pro Leu Arg Ala Phe Asp Met Ile Asn Arg Phe Ile Tyr Gly Lys Gly Trp Asp Pro Tyr Val Gly <210> 5 <211> 750 <212> DNA
<213> Homo sapiens <400> 5 acgatgtgtt gaccggctgc cgtttgagga ctttggtcac ccagactaga caccttctgt 60 gctcatgttt ggaaagctga aagggaagga cagctgtgcc ctcctgggag ctcatgtgtc 120 cctggcgctg tgctagcttt cctttacagc tgtttacaga caaggcaggc ctgaggcaga 180 tggccactgc tcttgtgatg tttgctcaga ggaatatgaa cattttattt ttgaaaaggg 240 atgatgtggt ttttgccagg tgtttataat taatccttta atattatggt tattaacctc 300 ttaaacatga atgaattctt gattgtttta acacagtacc taagactaat gctttctgtg 360 gacaccactg agctctgcct caactccacc ctctgcgacc ggaggactat gcccctagta 420 actgctgtcg gtgtggacgc tgtgctggtt ctgttttcta aaggagcaga aggacaggtc 480 tctgagacag gatcgttgtc cctacaggag gaacagtggc cttgcttctt agacggtctt 540 cactgtgtgt tttaaaacaa caacaacaac aacaacaaca taaaactctt ttgacctgta 600 acttaaagat cataaacttc aggcaataat attttctgtg taagctttta aaattatttt 660 tggggatcat agcttgtttt attttgtgct ataaaattaa cagtattaaa tgacttatat 720 tcttagaata aaaaaaaaaa aaaaaaaaaa 750 <210> 6 <211> 89 <212> PRT
<213> Homo sapiens <400> 6 Met Val Ile Asn Leu Leu Asn Met Asn Glu Phe Leu Ile Val Leu Thr Gln Tyr Leu Arg Leu Met Leu Ser Val Asp Thr Thr Glu Leu Cys Leu Asn Ser Thr Leu Cys Asp Arg Arg Thr Met Pro Leu Val Thr Ala Val Gly Val Asp Ala Val Leu Val Leu Phe Ser Lys Gly Ala Glu Gly Gln Val Ser Glu Thr Gly Ser Leu Ser Leu Gln Glu Glu Gln Trp Pro Cys Phe Leu Asp Gly Leu His Cys Val Phe <210> 7 <211> 2156 <212> DNA
<213> Homo Sapiens <220>
<221> unsure <222> (1353) <400> 7 aagtgatcta cctgcctggg cctcccaagg tgctgggatt acgggtgtga gccaccgcgc 60 ccagcctatt cttttttgtt tgtgataatg gtcatcctaa tggacatgag gtagtgtcat 120 gtggttttga tttgcatgtc cctgataaat aatgatgttg accatctact catgtgcttg 180 ttggctattt gcatggcgtg tttggagaaa cgtctgttca agggctttgc cttttttttt 240 tgagacagar tcttactccg ttgccccarg ctggagtkcg gtggtgaggg gtgcactgca 300 acatccgcct tccaggttca agcgattctt gtgcctcagc ctcccaaaga gctgggatta 360 caaaagtgca gtttgcccat ttttaatcga ttttgttcct gagttggagt tttttgtata 420 ttcaggctgt taacccctta tgagatagat ggtttgcaca tagtctcttc cattctatag 480 gatatcattt ctgttaatag attcctttgc tgtgcagaaa ctttttagtt tgaggtcatc 540 ccatttgtct atttttactt tcgttgccct tgctgttggt gtcatgttca agaaatcatt 600 gccaagacca atgtcgtgaa gtctttccct ttgttttctt ctaagggttt tacagtttca 660 agtctgtgtt tgggtcttgc atcggttttg agttagtttt tgtgtatgat gtaaggtaag 720 ggtctatctt tatttgcaag tggatatcca gttttcccag cgctgcatat tgaagagacc 780 atcctttccc cattgtgcaa gaagttcttg tcacccttgt tgaaggtcat ctgtctgtca 840 ttgtcatttc tggccctgtg ctgtcctgtc ctgtcctgtc ctgtcctgtt ctgttctgtt 900 ggtctgtagg tctgtcttta tgtcagcacc atactggctg ttggactttt taattctttt 960 cttgacagtg gtaatttatt tgcttctttt tcttattagt ccctttgcct actttaaata 1020 attaattttg ttaattttta gttttctgtt attttagttc attaatttca ttgcttcctt 1080 tatttattta tttatttttt ttgagatgga gtcttgctct gtcactcagg ctggagtgca 1140 gtggcacgat ctcagctcac tgcaacctcc acctcccagg ttcaagtgat tctcctgtct 1200 cagtctcctg agtagctggg attacaggca cttgccacca tgcccggcta attttttgta 1260 ttttttagta gagacggggt ttcgctgtgt tgcccgggct ggtttcaaac ttctgagctc 1320 aggcaatcca cctgcctcgg cctcccaaag tgntaggatt acaggtgtga gccaccacgc 1380 ctgacccatt gctgccttaa atacacaaag cgcttgagtt aataaagtta cctgaaggat 1440 tgaactttaa tttctaacag cgtttggagg tgaggggact acttgttttt gctcattttt 1500 agtttttttt tttttgcact tggggtcasa tggcatgtca tatgtgctgt tacctgaaat 1560 atattgaggg tttctttgtt ctatcatacm tggtcatttt cataactgtc ccacagacac 1620 tggagaagca tgatgactcc atggggtaca gaatttagaa catccttgtc agattgagtc 1680 tatggtgatg tgtcttaagt cgtcccttag tctttttttt cctaatcagt ctgtcaaatt 1740 tcagagaacc atgttaaaat cccctattat tgtggttttg aaggttgttt ccagtgtttt 1800 tccttcattt aattcttcct ctgtcgctgt gcgcctgcag attccaggct gcttgacatg 1860 ggttcctttc catatgggag tgagccagca gacagcccta cagatcgtac acacgttttc 1920 caaaactaac aatggaacag gcggcaaacc tatgccaata tactagaaat tgcagattaa 1980 atagatgaaa tattctaaac tggagtttac ataatgaaca taagagtaat cagagaatct 2040 gactcatttt agatgtgtgt gtgtgtgtat atatatgtgt gtgtgtgtga aaaacattga 2100 ctataataaa aataatctcg agttcaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaa 2156 <210> 8 <211> 94 <212> PRT
<213> Homo sapiens <400> 8 Met Val Met Cys Leu Lys Ser Ser Leu Ser Leu Phe Phe Pro Asn Gln Ser Val Lys Phe Gln Arg Thr Met Leu Lys Ser Pro Ile Ile Val Val Leu Lys Val Val Ser Ser Val Phe Pro Ser Phe Asn Ser Ser Ser Val 35 ~ 40 45 Ala Val Arg Leu Gln Ile Pro Gly Cys Leu Thr Trp Val Pro Phe His Met Gly Val Ser Gln Gln Thr Ala Leu Gln Ile Val His Thr Phe Ser Lys Thr Asn Asn Gly Thr Gly Gly Lys Pro Met Pro Ile Tyr <210> 9 <211> 3941 <212> DNA
<213> Homo sapiens <220>
<221> unsure <222> (2895) <400> 9 cagacacaga gatcagaatt ccaggaaatg atcttccagt gcgttctggg tcagttatgg 60 tgactgtaaa taccgtcatc acagctggcc ctcaaaataa cgcaataata acatatttac 120 ataatgacat attatgactg taagtgcagt cagccccatc tggggctgag gcgggggccc 180 tgctgtgcac tctcccccca gctatcccac cgggccaggg gtgggcctca gggttgtgct 240 gggagccgca gggcctgaag gggcctcggc tgtacgggga tgagactcgc aggggagagg 300 gcagaggccg gtgacctggc gaggacttgc ccaggagatt ggagctcctt gcttctgcgc 360 cacgcggatg ccccacgctg gtctcagctg ggttgttggc tctgagtggt catctcgttg 420 ctgccatatt ttcttgcttc attgaatttc actgtgctcc agcctgggca acacagccgg 480 actctgtttc aaaaaaaaaa attttttttt tccaagatag gatggtagag aaaatacctc 540 ctgccatgtc ctgctatgaa tacagctttg tatttctctc tctagttttg tcagttttgg 600 cttttcagat tttgaagcgt gtttgtgggc tgaatcttgc ccttatcacc catttctagg 660 atgctttttg ctccactcat tctttgtctt gcttcacttg actttgaact gtatactttt 720 ttccatcgtt ttactttcag tatcttcata catgtatgtt tttgtacgcc tctcttagaa 780 cagtgtatgg ttttgtaaaa attcagcctg tagcttttac ctgcctcctt catgaccttt 840 ataatcccct tggttctcag cctgccactc acaggacttt tccctgtgct gcgttccmag 900 tgccccctcc ccgcccccac ctgtgctttt tgttggatta gtagaattgc ttttgtcatt 960 ccattgtttt catatatttg tttgggacat tttacttttt tctgttaacg cttaccctag 1020 aaattagaaa tgacaccacg tattcttagc gaagtccagt tttcagcatt ttgtccttat 1080 tggacaatag caaggatatt agaacgtgtt ggttccgcgt gcttccgtct tgagttatgt 1140 gctgctattg tcggatattt tgtcttagat gtacgtactt tcctgttcat tgtggtatgt 1200 gtaatttgcg ttactttgaa ttttccacgt ttttactttc tttgtctctc atcacttact 1260 gcttttggga ccccccccat cggggttcac attccctctc cctagagcac actcccttgg 1320 atttcctcga gtggggtctg ctgcggtgaa gctttcccat tttatgtgca gattattttc 1380 agagggtata tagaattcag gcagctgttt cgttgtagca cattaaaaat attttcccac 1440 ttcctccttg cttctgttgt tgcttttgag tgttacctct gagtctgcct gtgctccctg 1500 gaaacggccc gggtttccca ccccctgccc aggtttgctc cttccgtggt ttttctgtca 1560 ttatcacgct cacgtgtttc cctcggtcac cccctctgca attttcacac gtcttttccc 1620 tctctctttg cttcattacc tttggcccgc ctgccagctg ctgattctct ctgaagatgt 1680 ctctaaatga cttttaactg tgatttgtgg aattcttatt gtggagtttt gcgtcttttc 1740 aggtgtaggt tttttgtctc gcgtgtttcc acgtctgctt gtagcgcttt ccgcttcgcc 1800 gttccctgcg gcccttcctt ccgtgcccgg tgttcatcct cttgaatgct cttttcctgc 1860 ctgtttggct gggtgtgtct gagttgcaac ctgagcgggt ttctttgtct tcttacttgt 1920 ctggtattgt gttctctcgg gacgttgcgt ttgaggggtc gcacctcaga gcaagccgag 1980 gtctgggcta agcctgtgct ttggcaggca ggaccttagt ttgccttttc tgggcacctg 2040 aggagagggt agcagcagcc tggggtctcc ttgactcacg gtcagcagtg agggtttcct 2100 ggcctgttgg gtggctggag cttggctgca ttccccactg agagagggag gtgcgcacct 2160 tctcctccct ggagtggcct tccaggtgcc ctctcagagc tgctcatcag ggctgtgcct 2220 ttgtcagcac caagcctcag cccttgtccc tgctgccact gaaggctcaa aacaacactg 2280 cacagccttg tgtgtcctct gtgtgtcggc agtttccccc ggctctgcag cagcccaggc 2340 caggtagcct ctggagggag tggtggagga gcacgggcat cctggccgcc gctgtgttgg 2400 ggacagaccc tggggcctgg aaagggaggt gaggccccgt gggggctgct gcaccacagg 2460 caagagagca agagacagca gaggccggcc aggtggtggc acagccgcta gggaccaggc 2520 cggcctgtgg aggtattggg atggggacca gcggacttgc tggcagaggg gcctcagggc 2580 tgcaggcttc ttggactgag ccactgggag gacggagttg accttctttg agacagaaaa 2640 agtgtgcatc ccggggctgc ctgtgaaagc tcatctctaa agtgtgtgtt gttcttccag 2700 ccaccccttt gctgtgaagt tgcttgcgct ctgtaagaaa gaaatcaaga attcaaaaga 2760 tatccagaag ctcctgtcag gcatcgcagt gtgagtttca agtgctactg gccttagacg 2820 gaatggcagg gcgcagcctc ccttggctga gggcaggagt ccacggctcc aggcgggaga 2880 ggagcagtta gtgtnactcc tcaagctaac ctaagatcgt gcattccaat gttcaaagca 2940 gtcgcaatgg gaggtgaggc agcccaggtg ctggtggagg gagttcccgc gggaacaggc 3000 gagctctgcc tctgctgccc tcgcgctctg ccctggcggg aggggaggct ccggaaagga 3060 gctgcgtggt caggggctgc ctccccgatt ctcctgtgtg ccctgggggt cgctgttgag 3120 tgccttgctc tgcggcgctc aggtggacac tgggcaggtg cgccagccag cgataggcac 3180 cttggctgct ctgtggctcc ttgaggtggg ggtcctcatg gcagggcgag cggccctgca 3240 ggagatcctc tgtgaggcgt cctcacttcc cacagtgact ttccaagtgc gacactcgcg 3300 tgtgtaggca cagtgcagat gtgcgcacac acacacctcc ggcttggggc cccaggcccg 3360 cactgtgctc acggatctgc tctgcccagg ttctgcggga tggtgcagtt ccccggcgaa 3420 cgtgaggagg caggccctcc tgcagctgtg tctgctcctc tgccaccgtt tccsgctgat 3480 ccggaagacc acggccagcc aggtgtacga gacattgctc acctacagtg acktcgtggg 3540 cgcggatgtg ctggacgagg tggtgactgt gctcagtgac actgsgtgga cgcagagctt 3600 gcagtggtga gagagcagcg caaccgtctg tgtgaccttc tgggcgtacc caggccccag 3660 ytggtgcccc agcctggtgc ctgctgaagc cagtcctgga gcccatacct cacccctgcc 3720 tggtgaggat gtcttgttcc tgagggaggc cggtgtggaa agcctcgcac agtggtgcct 3780 ccagctgttg aagggtagcg ctggcccttg gaggctggca ctagctgaca gcttttcctc 3840 tctgcacctg cgctctggtg acttggggtg gacgcctctg ccttcacttg aacacaaatg 3900 tgcttcctat aaaatcatgt accaagaaaa aaaaaaaaaa a 3941 <210> 10 <211> 70 <212> PRT
<213> Homo Sapiens <400> 10 Met Cys Cys Tyr Cys Arg Ile Phe Cys Leu Arg Cys Thr Tyr Phe Pro Val His Cys Gly Met Cys Asn Leu Arg Tyr Phe Glu Phe Ser Thr Phe Leu Leu Ser Leu Ser Leu Ile Thr Tyr Cys Phe Trp Asp Pro Pro His Arg Gly Ser His Ser Leu Ser Leu Glu His ThY Pro Leu Asp Phe Leu Glu Trp Gly Leu Leu Arg <210> 11 <211> 1779 <212> DNA
<213> Homo Sapiens <400> 11 ccaagttcca ggtctagaat tcaaattact aatttactgc ttctctctct ctaagcctca 60 gctccctgat ctagaccatg agatttacag taggagagta ccatgtttat ccccaaatac 120 ttaacagcta gggttttccc agactgaata ataataataa cttttttaaa attcagaagg 180 , tatcttcaag ttcttggctt gcttcttgta cattcaatat caaagaagag aaaacacact 240 atctgagagt acttcccatg cacctaataa gtgccaaagc cacctggtgc tagagccctt 300 caccaaaatg agcatcagcc ttgctttcag aaagcaggga ccacatatat atgatttaaa 360 aaaaatctgc gatcaacttt tctctaaaaa acccaaatat gctggggtac agaaagatca 420 atgcaaaagc aaaacatcct gtgcctgtcc tagaggtccc cagaggcagg atgccccgac 480 tcagaaagaa actcctaagc tggcctggcc aaagggagga agaacccagg gtgggtgtcg 540 taactcatct aaaaataacg atgtcatcag gcagatgtgc cattgtgctg gggctgggtg 600 ggtgtggcag gcccaccttg ggtatgcaaa gctctgacag tgtttcactt gctaccctcg 660 g gtctgcttac cacactccca gttctgctga ccttacggga aggctcatgc tgggttgact 720 cacggcaggc ctagagcact gtgagggatg tgtgaggaca agggtcacac cccagggtgg 780 catttccaag ccccatgcct ctggccatat cccatagggg ctctaggcct ctgttttccc 840 atctttaaaa taattggggg caatacctcc tatgatcttt ctgagaatta atagagattt 900 catggcaatt gcttagccct gcccagcaga gatagcaaat aatcaatcag ctccctttct 960 cctctgtctc ttgggtgttt tctactcctg gaaccccaga gcaagagagg accctgaaac 1020 atggcctaca tccaattctt tcattttgca tttgaggaaa tcgaggcaca tggctgcggt 1080 tctactctta ccaacccata tcaggtcatt gctctaacga ggcttaagga gcaataaccc 1140 gcctttcacg tggttcttac ggatacccag aaagatgact cagcttctcc agatttctga 1200 gaagactaag cataagtcag agagagtata gacaaaggaa aagggggcat aactgcaagg 1260 accccctcaa atgtgtgctg tggcagcatt ggtgggacag gggctgsaag agcaaaacag 1320 tagggatcac atcttggaga gtactcggga aggagtccaa saacgaccat ggatcctgga 1380 gctacaggtt gcaaccaaac tacaatcatt ccatttggcc tcaggatgtg gaagcacccc 1440 aaatgtgttt gcctcaaaaa gcaaagagga tgaggcccgg catggtagct caggcctgta 1500 atcccagcac tttgggaggc cgaggtgggc ggatcacttg agtccaggag ttcgagatca 1560 gcctgggcaa tgtagcaaca ccgcacctct acaaaaaata aaagaattaa ctgggcgtgg 1620 tggcgcatgc ctgtagtccc agctactctg gaggctgagg tgggaggatc ccttgagccc 1680 aggagatgga ggttgcagtg agctgagatg gcaccactgc actccagtct gggtgacaga 1740 gcaagaccca gactcaaaaa aaaaaaaaaa aaaaaaaaa 1779 <210> 12 <211> 109 <212> PRT
<213> Homo Sapiens <400> 12 Met Ser Ile Ser Leu Ala Phe Arg Lys Gln Gly Pro His Ile Tyr Asp Leu Lys Lys Ile Cys Asp Gln Leu Phe Ser Lys Lys Pro Lys Tyr Ala Gly Val Gln Lys Asp Gln Cys Lys Ser Lys Thr Ser Cys Ala Cys Pro Arg Gly Pro Gln Arg Gln Asp Ala Pro Thr Gln Lys Glu Thr Pro Lys Leu Ala Trp Pro Lys Gly Gly Arg Thr Gln Gly Gly Cys Arg Asn Ser Ser Lys Asn Asn Asp Val Ile Arg Gln Met Cys His Cys Ala Gly Ala Gly Trp Val Trp Gln Ala His Leu Gly Tyr Ala Lys Leu <210> 13 <211> 1170 <212> DNA
<213> Homo Sapiens <400> 13 agccgcgcgg ctgcgggggc gcaaataggg tcactgggcc gcttggcggt gtcgttgcgg 60 taccaggtcc gcgtgagggg ttcgggggtt ctgggcaggc acaatggcgt ctcgagcagg 120 cccgcgagcg gccggcaccg acggcagcga ctttcagcac cgggagcgcg tcgccatgca 180 ctaccagatg agtgtgaccc tcaagtatga aatcaagaag ctgatctacg tacatctggt 240 catatggctg ctgctggttg ctaagatgag cgtgggacac ctgaggctct tgtcacatga 300 tcaggtggcc atgccctatc agtgggaata cccgtatttg ctgagcattt tgccctctct 360 cttgggcctt ctctcctttc cccgcaacaa cattagctac ctggtgctct ccatgatcag 420 catgggactc ttttccatcg ctccactcat ttatggcagc atggagatgt tccctgctgc 480 acagcagctc taccgccatg gcaaggccta ccgtttcctc tttggttttt ctgccgtttc 540 catcatgtac ctggtgttgg tgttggcagt gcaagtgcat gcctggcagt tgtactacag 600 caagaagctc ctagactctt ggttcaccag cacacaggag aagaagcata aatgaagcct 660 ctttggggtg aagcctggac atcccatcga atgaaaggac actagtacag cggttccaaa 720 atcccttctg gtgattttag cagctgtgat gttggtacct ggtgcagacc aggccaaagt 780 tctggaaagc tccttttgcc atctgctgag gtggcaaaac tataatttat tcctggttgg 840 ctagaactgg gtgaccgaca gctatgaaac aaatttcagc tgtttgaagt tgaactttga 900 ggtttttctt taagaatgag cttcgtcctt gcctctactc ggtcattctc cccatttcca 960 tccattaccc cttagccatt gagactaaag gaaataggga ataaatcaaa ttacttcatc 1020 tctaggtcac gggtcaggaa acatttgggc agctgctccc ttggcagctg tggtctcctc 1080 tgcaaagcat tttaattaaa aacctcaata aagatggccc tgcccacaaa aaaaaaaaaa 1140 aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa 1170 <210> 14 <211> 183 <212> PRT
<213> Homo sapiens <400> 14 Met Ala Ser Arg Ala Gly Pro Arg Ala Ala Gly Thr Asp Gly Ser Asp Phe Gln His Arg Glu Arg Val Ala Met His Tyr Gln Met Ser Val Thr Leu Lys Tyr Glu Ile Lys Lys Leu Ile Tyr Val His Leu Val Ile Trp Leu Leu Leu Val Ala Lys Met Ser Val Gly His Leu Arg Leu Leu Ser His Asp Gln Val Ala Met Pro Tyr Gln Trp Glu Tyr Pro Tyr Leu Leu Ser Ile Leu Pro Ser Leu Leu-Gly Leu Leu Ser Phe Pro Arg Asn Asn Ile Ser Tyr Leu Val Leu Ser Met Ile Ser Met Gly Leu Phe Ser Ile Ala Pro Leu Ile Tyr Gly Ser Met Glu Met Phe Pro Ala Ala Gln Gln Leu Tyr Arg His Gly Lys Ala Tyr Arg Phe Leu Phe Gly Phe Ser Ala Val Ser Ile Met Tyr Leu Val Leu Val Leu Ala Val Gln Val His Ala Trp Gln Leu Tyr Tyr Ser Lys Lys Leu Leu Asp Ser Trp Phe Thr Ser Thr Gln Glu Lys Lys His Lys <210> 15 <211> 1127 1~

<212> DNA
<213> Homo sapiens <400> 15 ctcgccgcag aagtatctcc gaatggagcc atcccccttc ggcgacgtct cctcccgcct 60 caccacagaa caaattctgt acaacataas acaagagtat aaacgaatgc agaagagaag 120 acatttagaa acgagtttcc aacagacaga tccgtgttgt acttctgatg cacagccaca 180 tgcatttctc ctcagtggac cagcttcacc agggacttca tctgcagcat cctcaccatt 240 aaaaaaagaa cagcccttat ttactctacg gcaggttggg atgatctgtg aacgtCtgtt 300 gaaagaacgt gaagagaaag ttcgagaaga atatgaagaa atattgaaca caaaacttgc 360 agaacaatat gatgcgtttg tgaagtttac gcatgatcaa ataatgcgac gatatggaga 420 acagcctgct agctatgttt catgaatcac gtatcctgca tttgtgggct gccttgttcc 480 ttgttgagtt gttgcaagag gtcccaatta tgacatgcag caatgccaat accccttctg 540 tgaatacagg ttatttcaag ctttcgtcag tggcaaccac tcttaggcag cagcaactgg 600 ttttggaaat ttccctgatg tcagtaccac ctggatgtgg acctttgcta cctgtattaa 660 taccagtggc ctcattttgc tgtatcatta caatttggct tcttatatta atgtttgaaa 720 aggattaaag ctggtattct agaacatgcc cttcactggt tgtgtaaata aaactgtaga 780 atgacacttc agatgaagtt agtgtgattt taattgtgca ctacaaccga gctgtaacca 840 gttactaatt ttagaatgta atcccaggac aatattaagc aaatagcctg cagtgcttcc 900 tgtgaaatag tgaaggagga gggcatttct gtattccagg acttcttggg gtttcagaat 960 gggtttgtat gatttttttt tttttgtagt tttatttatt ctatcagtct ttttaacaaa 1020 tgtttattgc tgcatttttt tttttccagt gtatcattgt tttactgccc ttgtagtact 1080 ggaatttagt tggaagaata aaacatttac ttctaaaaaa aasaaaa <210> 16 <211> 140 <212> PRT
<213> Homo sapiens <400> 16 Met Glu Pro Ser Pro Phe Gly Asp Val Ser Ser Arg Leu Thr Thr Glu Gln Ile Leu Tyr Asn Ile Lys Gln Glu Tyr Lys Arg Met Gln Lys Arg Arg His Leu Glu Thr Ser Phe Gln Gln Thr Asp pro Cys Cys Thr Ser Asp Ala Gln Pro His Ala Phe Leu Leu Ser Gly Pro Ala Ser Pro Gly Thr Ser Ser Ala Ala Ser Ser Pro Leu Lys Lys Glu Gln Pro Leu Phe Thr Leu Arg Gln Val Gly Met Ile Cys Glu Arg Leu Leu Lys Glu Arg Glu Glu Lys Val Arg Glu Glu Tyr Glu Glu Ile Leu Asn Thr Lys Leu Ala Glu Gln Tyr Asp Ala Phe Val Lys Phe Thr His Asp Gln Ile Met Arg Arg Tyr Gly Glu Gln Pro Ala Ser Tyr Val Ser <210> 17 <211> 806 <212> DNA
<213> Homo sapiens <400> 17 gtgtatcttc agaggcagca ggggccagtg tgccacatct tgccccagtc ctgaaaggat 60 agatggtatt tggcctgtga cccttggctg aggagccatg gtccggctct gccaggccct 120 gctgctgtta gtggccactg tggcccttgc atccagaaga ttccaagcct ggggctcaac 180 aaargtggtg aggacattcc aagatatccc tcaaaactac gtctatgtkc arcakgcact 240 ctggttcgcc atagaaggag tataacaagg ccagctttag tataacaagt tcagctttag 300 ggtgctgaag gttctgaaga gccasgarca ggtgacagat agtttggagt actatattga 360 ggtcaaaatt gcccgaacar tttgcaagaa aatttcagaa gatgaaaact gtgcatttca 420 agaggatccc aaaatgcaaa aggtggtttt ttgtaytttt attgttgcat ctaaaccatg 480 gaaatttgaa ctcaccatgy tgraaacaat gcaaagatat gtagttatct tctmgtgtgt 540 tctgccacac tcatttccat tttaaagaag aagcaaagac ayttgcaaga aytagaacaa 600 cacagttaac ccattaactt catttgtttg gcctttttgc atttttgtgt gttcttcatg 660 ggctgatgtt gaaaatccat gatgtgtttt gacagcattg catagcctat tcttgctgga 720 tacttcccct actagctggg ataatctgyt gcaataaatg gaagtggttt cttacacstc 780 aaaaaaaaaa aaaaaaaaaa aaaaaa 806 <210> 18 <211> 55 <212> PRT
<213> Homo sapiens <220>
<221> UNSURE
<222> (46) <400> 18 Met Val Arg Leu Cys Gln Ala Leu Leu Leu Leu Val Ala Thr Val Ala Leu Ala Ser Arg Arg Phe Gln Ala Trp Gly Ser Thr Lys Val Val Arg Thr Phe Gln Asp Ile Pro Gln Asn Tyr Val Tyr Val Gln Xaa Ala Leu Trp Phe Ala Ile Glu Gly Val <210> 19 <211> 1783 <212> DNA
<213> Homo sapiens <400> 19 tccccacccc ccttatgtct cagccgaacc taccctaatc cagcccacgc cacaatggtg 60 ggacaggttc cccagtccct atgtggtctt atttttaccc ttgcactccc tgtagaccat 120 caattctaca ccctaattac aaaatcatat ccacctctgc ctggcagaag gtgttatgct 180 tttctggctc gcctaccatc cacacatccc tacacctcac caccggatcc tcttttcttt 240 ccttccatcc aattcctggc ttccccgctg ccaactctgc tctctatgtc tccagtttaa 300 aggtgccccc tggaaaaaat gtaacaattc cctcacctgt gactggtacc tgacagccac 360 cacaccgggg cagcaatggc taacggttga caaagacaat ttctttctct ctccaaaacc 420 aaacagcctt catcaactcc ctagccaaga ctccctatca ggcccttaca ggtgccgctc 480 tggctggcag ttacccmatt tgggaaaacg aaaataccct atcatggcta cctaccttca 540 cctacaactt ctgcctgtcc acccccagtc tcttcttttt gtgtgataca aactgatatc 600 tttgcctacc agccaactgg tcaggaactt gcaccctggt ctttcaggct ccaaccatca 660 acatcctacc ecctaaccaa actattctaa tttctgtaga agcctctatc tcctcttcac 720 WO 99/3b512 PCT/US99/00550 ccataagaaa taaatgggct ctacatctca tcaccctgct aacaggatta ggcatcactg 780 ctgcacttgg cactggasta gcaggcataa ccacctcaat cacctcatac caaacactat 840 tcacaaccct ttctaacacc gtagaagata tgcacacttc cattaccagt ctccaacgac 900 aattagactt cctcgtggga gtcatccttc aaaactggag agtcctggac ctcctaacca 960 ctgagaaagg gggtacctgc atatacctcc aggaagaatg ctgtttctgt gttaatgaat 1020 ctggcattgt tcatatcgca gttcgtaggc ttcatgacag ggctgcagag ctttgacatc 1080 aagtcgctga ctcctggtgg caaggatcat cccttctaag atggataccc tgggttgccc 1140 ccttcctagg acccctgatc ttcctcttcc tgttactaat gattgggcca tgcatattta 1200 accttgtatc ccgcttcatt tcccaaaggc tgaattgttt tatccaggca agcatgcaaa 1260 aacacattga taatatattt cacctttgcc acgtctaata ccagagccta cgaggaaacc 1320 attcggaagc tccagaaccc aggccctaat cacaacgccc ctatccagca ggaagcagcc 1380 agatgatyaa mgacgccctt tttccttttt atactaaagt aagaaataag aatgttagcc 1440 caaactgcay tattttgcag acccctacca ttttacaaac tggtcagagt ggaaaattcc 1500 accagggcct gagctgtgag aaacatcctg tcaggcaggt cccaggccta acccctggst 1560 gcactaaatt ccttcattat cagcagccaa acacaccgcc cccaccccat tttcacaaca 1620 atcccagacc tctcctgccc gggactgtaa ctggtccagc ctgtaagcgg gaagggggct 1680 ctggcactag stggtacccc ctctccgcag gtctttctcc caataaatct gtgttgccct 1740 tgraaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaa 1783 <210> 20 <211> 136 <212> PRT
<2I3> Homo sapiens <220>
<221> UNSURE
<222> (108) <400> 20 Met Leu Phe Trp Leu Ala Tyr His Pro His Ile Pro Thr Pro His His Arg Ile Leu Phe Ser Phe Leu Pro Ser Asn Ser Trp Leu Pro Arg Cys Gln Leu Cys Ser Leu Cys Leu Gln Phe Lys Gly Ala Pro Trp Lys Lys Cys Asn Asn Ser Leu Thr Cys Asp Trp Tyr Leu Thr Ala Thr Thr Pro 50 55 ~ 60 Gly Gln Gln Trp Leu Thr Val Asp Lys Asp Asn Phe Phe Leu Ser Pro Lys Pro Asn Ser Leu His Gln Leu Pro Ser Gln Asp Ser Leu Ser Gly Pro Tyr Arg Cys Arg Ser Gly Trp Gln Leu Pro Xaa Leu Gly Lys Arg Lys Tyr Pro Ile Met Ala Thr Tyr Leu His Leu Gln Leu Leu Pro Val His Pro Gln Ser Leu Leu Phe Val <210> 21 <211> 19 <212> DNA
<213> Artificial Sequence <220>

<223> oligonucleotide <400> 21 ccgtgatacc gaaatgtcc 1g <210> 22 <211> 29 <212> DNA

<213> Artificial Sequence <220>

<223> oligonucleotide <220>

<221> misc_feature <222> (2 ) <223> biotinylated phosphoaramidite residue <400> 22 gnaacaatca ccttccacat ggcaccaac <210> 23 <211> 29 <212> DNA

<213> Artificial Sequence <220>

<223> oligonucleotide <220>

<221> misc_feature <222> (2) <223> biotinylated phosphoaramidite residue <400> 23 gngttgaggc agagctcagt ggtgtccac <210> 24 <211> 29 <212> DNA

<213> Artificial Sequence <220>

<223> oligonucleotide <220>

<221> misc_feature <222> (2) <223> biotinylated phosphoaramidite residue <400> 24 ancgtgtgta cgatctgtag ggctgtctg 29 <210> 25 <211> 29 <212> DNA

<213> Artificial Sequence <220>

<223> oligonucleotide <220>

<221> misc_feature <222> (2) <223> biotinylated phosphoaramidite residue <400> 25 gnagcacgcg gaaccaacac gttctaata <210> 26 <211> 29 <212> DNA

<213> Artificial Sequence <220>

<223> oligonucleotide <220>

<221> misc_feature <222> (2) <223> biotinylated phosphoaramidite residue <400> 26 anatcaggga gctgaggctt agagagaga 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 gngaaaggag agaaggccca agagagagg <210> 28 <211> 29 <212> DNA

<213> Artificial Sequence <220>

<223> oligonucleotide <220>

<221> misc_feature <222> (2) <223> biotinylated phosphoaramidite residue <400> 28 gntgccactg acgaaagctt gaaataacc 29 <210> 29 <211> 20 <212> DNA
<213> Artificial Seguence <220>
<223> oligonucleotide <400> 29 ggctctacat ctcatcaccc ~ 20 <210> 30 .
<211> 130 <212> PRT
<213> Homo sapiens <400> 30 Met Asp Asp Met Leu Ser Thr Arg Ser Ser Thr Leu Thr Glu Asp Gly Ala Lys Ser Ser Glu Ala Ile Lys Glu Ser Ser Lys Phe Pro Phe Gly Ile Ser Pro Ala Gln Ser His Arg Asn Ile Lys Ile Leu Glu Asp Glu Pro His Ser Lys Asp Glu Thr Pro Leu Cys Thr Leu Leu Asp Trp Gln Asp Ser Leu Ala Lys Arg Cys Val Cys Val Ser Asn Thr Ile Arg Ser Leu Ser Phe Val Pro Gly Asn Asp Phe Glu Met Ser Lys His Pro Gly Leu Leu Leu Ile Leu Gly Lys Leu Ile Leu Leu His His Lys His Pro Glu Arg Lys Gln Ala Pro Leu Thr Tyr Glu Lys Glu Glu Glu Gln Asp Gln Gly <210> 31 <211> 111 <212> PRT
<213> Homo sapiens <400> 31 Met Leu Gly Tyr Arg Lys Ile Asn Ala Lys Ala Lys His Pro Val Pro Val Leu Glu Val Pro Arg Gly Arg Met Pro Arg Leu Arg Lys Lys Leu Leu Ser Trp Pro Gly Gln Arg Glu Glu Glu Pro Arg Val Gly Val Val Thr His Leu Lys Ile Thr Met Ser Ser Gly Arg Cys Ala Ile Val Leu Gly Leu Gly Gly Cys Gly Arg Pro Thr Leu Gly Met Gln Ser Ser Asp 65 70 75 g0 Ser Val Ser Leu Ala Thr Leu Gly Leu Leu Thr Thr Leu Pro Val Leu g5 90 95 Leu Thr Leu Arg Glu Gly Ser Cys Trp Val Asp Ser Arg Gln Ala <210> 32 <211> 104 <212> PRT
<213> Homo Sapiens <400> 32 Met Glu Asn Ser Leu Leu Ala Met Phe His Glu Ser Arg Ile Leu His Leu Trp Ala Ala Leu Phe Leu Val Glu Leu Leu Gln Glu Val Pro Ile Met Thr Cys Ser Asn Ala Asn Thr Pro Ser Val Asn Thr Gly Tyr Phe Lys Leu Ser Ser Val Ala Thr Thr Leu Arg Gln Gln Gln Leu Val Leu Glu Ile Ser Leu Met Ser Val Pro Pro Gly Cys Gly Pro Leu Leu Pro 65 70 75 g0 Val Leu Ile Pro Val Ala Ser Phe Cys Cys Ile Ile Thr Ile Trp Leu 85 90 g5 Leu Ile Leu Met Phe Glu Lys Asp

Claims (41)

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 427 to nucleotide 1146;
(c) a polynucleotide comprising the nucleotide sequence of SEQ ID
NO:1 from nucleotide 589 to nucleotide 1146;
(d) a polynucleotide comprising the nucleotide sequence of the full-length protein coding sequence of clone bd577 1 deposited under accession number ATCC 98631;
(e) a polynucleotide encoding the full-length protein encoded by the cDNA insert of clone bd577_1 deposited under accession number ATCC 98631;
(f) a polynucleotide comprising the nucleotide sequence of the mature protein coding sequence of clone bd577 1 deposited under accession number ATCC 98631;
(g) a polynucleotide encoding the mature protein encoded by the cDNA insert of clone bd577_1 deposited under accession number ATCC 98631;
(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 having biological activity, the fragment comprising eight consecutive amino acids of SEQ ID NO:2;
(j) a polynucleotide which is an allelic variant of a polynucleotide of (a)-(g) above; and (k) a polynucleotide that hybridizes under stringent conditions to any one of the polynucleotides specified in (a)-(i).
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 bd577 1 deposited under accession number ATCC 98631.
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) fragments of the amino acid sequence of SEQ ID NO:2, each fragment comprising eight consecutive amino acids of SEQ ID NO:2; and (c) the amino acid sequence encoded by the cDNA insert of clone bd577_1 deposited under accession number ATCC 98631;
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. A process for producing an isolated polynucleotide, wherein the process is 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 NO:1, but excluding the poly(A) tail at the 3' end of SEQ ID NO:1; and (ab) the nucleotide sequence of the cDNA insert of clone bd577_1 deposited under accession number ATCC 98631;
(ii) hybridizing said probe(s) to human genomic DNA in conditions at least as stringent as 4X SSC at 65 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 NO:1, but excluding the poly(A) tail at the 3' end of SEQ ID NO:1; and (bb) the nucleotide sequence of the cDNA insert of clone bd577 1 deposited under accession number ATCC 98631;
(ii) hybridizing said primer(s) to human genomic DNA in conditions at least as stringent as 4X SSC at 65 degrees C;
(iii) amplifying human DNA sequences; and (iv) isolating the polynucleotide products of step (b)(iii);
wherein at least one isolated polynucleotide comprises a nucleotide sequence selected from the group consisting of:
(v) 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;
(w) a nucleotide sequence corresponding to the cDNA sequence of SEQ ID
NO:1 from nucleotide 427 to nucleotide 1146, and extending contiguously from a nucleotide sequence corresponding to the 5' end of said sequence of SEQ ID
NO:1 from nucleotide 427 to nucleotide 1146, to a nucleotide sequence corresponding to the 3' end of said sequence of SEQ ID NO:1 from nucleotide 427 to nucleotide 1146; and (x) a nucleotide sequence corresponding to the cDNA sequence of SEQ ID
NO:1 from nucleotide 589 to nucleotide 1146, and extending contiguously from a nucleotide sequence corresponding to the 5' end of said sequence of SEQ ID
NO:1 from nucleotide 589 to nucleotide 1146, to a nucleotide sequence corresponding to the 3' end of said sequence of SEQ ID NO:1 from nucleotide 589 to nucleotide 1146.
13. An isolated polynucleotide produced according to the process of claim 12.
14. An isolated polynucleotide comprising the polynucleotide of claim 13.
15. 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 95 to nucleotide 1522;
(c) a polynucleotide comprising the nucleotide sequence of SEQ ID
NO:3 from nucleotide 161 to nucleotide 1522;
(d) a polynucleotide comprising the nucleotide sequence of the full-length protein coding sequence of clone bv280_3 deposited under accession number ATCC 98631;
(e) a polynucleotide encoding the full-length protein encoded by the cDNA insert of clone bv280_3 deposited under accession number ATCC 98631;
(f) a polynucleotide comprising the nucleotide sequence of the mature protein coding sequence of clone bv280_3 deposited under accession number ATCC 98631;
(g) a polynucleotide encoding the mature protein encoded by the cDNA insert of clone bv280_3 deposited under accession number ATCC 98631;
(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 having biological activity, the fragment comprising eight consecutive amino acids of SEQ ID NO:4;

(j) a polynucleotide which is an allelic variant of a polynucleotide of (a)-(g) above; and (k) a polynucleotide that hybridizes under stringent conditions to any one of the polynucleotides specified in (a)-(i).
16. A protein comprising an amino acid sequence selected from the group consisting of:
(a) the amino acid sequence of SEQ ID NO:4;
(b) fragments of the amino acid sequence of SEQ ID NO:4, each fragment comprising eight consecutive amino acids of SEQ ID NO:4; and (c) the amino acid sequence encoded by the cDNA insert of clone bv280_3 deposited under accession number ATCC 98631;
the protein being substantially free from other mammalian proteins.
17. A process for producing an isolated polynucleotide, wherein the process is 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 NO:3, but excluding the poly(A) tail at the 3' end of SEQ ID NO:3; and (ab) the nucleotide sequence of the cDNA insert of clone bv280_3 deposited under accession number ATCC 98631;
(ii) hybridizing said probe(s) to human genomic DNA in conditions at least as stringent as 4X SSC at 65 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 NO:3, but excluding the poly(A) tail at the 3' end of SEQ ID NO:3; and (bb) the nucleotide sequence of the cDNA insert of clone bv20-3 deposited under accession number ATCC 98631;
(ii) hybridizing said primer(s) to human genomic DNA in conditions at least as stringent as 4X SSC at 65 degrees C;
(iii) amplifying human DNA sequences; and (iv) isolating the polynucleotide products of step (b)(iii);
wherein at least one isolated polynucleotide comprises a nucleotide sequence selected from the group consisting of:
(v) a nucleotide sequence corresponding to the cDNA sequence of SEQ ID
NO:3, and extending contiguously from a nucleotide sequence corresponding to the 5' end of SEQ ID NO:3 to a nucleotide sequence corresponding to the 3' end of SEQ ID NO:3, but excluding the poly(A) tail at the 3' end of SEQ ID NO:3;
(w) a nucleotide sequence corresponding to the cDNA sequence of SEQ ID
NO:3 from nucleotide 95 to nucleotide 1522, and extending contiguously from a nucleotide sequence corresponding to the 5' end of said sequence of SEQ ID
NO:3 from nucleotide 95 to nucleotide 1522, to a nucleotide sequence corresponding to the 3' end of said sequence of SEQ ID NO:3 from nucleotide 95 to nucleotide 1522;
and (x) a nucleotide sequence corresponding to the cDNA sequence of SEQ ID
NO:3 from nucleotide 161 to nucleotide 1522, and extending contiguously from a nucleotide sequence corresponding to the 5' end of said sequence of SEQ ID
NO:3 from nucleotide 161 to nucleotide 1522, to a nucleotide sequence corresponding to the 3' end of said sequence of SEQ ID NO:3 from nucleotide 161 to nucleotide 1522.
18. 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 286 to nucleotide 552;
(c) a polynucleotide comprising the nucleotide sequence of SEQ ID
NO:5 from nucleotide 475 to nucleotide 552;

(d) a polynucleotide comprising the nucleotide sequence of the full-length protein coding sequence of clone co315_3 deposited under accession number ATCC 98631;
(e) a polynucleotide encoding the full-length protein encoded by the cDNA insert of clone co315-3 deposited under accession number ATCC 98631;
(f) a polynucleotide comprising the nucleotide sequence of the mature protein coding sequence of clone co315_3 deposited under accession number ATCC 98631;
(g) a polynucleotide encoding the mature protein encoded by the cDNA insert of clone co315_3 deposited under accession number ATCC 98631;
(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 acid sequence of SEQ ID NO:6 having biological activity, the fragment comprising eight consecutive amino acids of SEQ ID NO:6;
(j) a polynucleotide which is an allelic variant of a polynucleotide of (a)-(g) above; and (k) a polynucleotide that hybridizes under stringent conditions to any one of the polynucleotides specified in (a)-(i).
19. A protein comprising an amino acid sequence selected from the group consisting of:
(a) the amino acid sequence of SEQ ID NO:6;
(b) fragments of the amino acid sequence of SEQ ID NO:6, each fragment comprising eight consecutive amino acids of SEQ ID NO:6; and (c) the amino acid sequence encoded by the cDNA insert of clone co315_3 deposited under accession number ATCC 98631;
the protein being substantially free from other mammalian proteins.
20. A process for producing an isolated polynucleotide, wherein the process is 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 NO:5, but excluding the poly(A) tail at the 3' end of SEQ ID NO:5; and (ab) the nucleotide sequence of the cDNA insert of clone co315_3 deposited under accession number ATCC 98b31;
(ii) hybridizing said probe(s) to human genomic DNA in conditions at least as stringent as 4X SSC at 65 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 NO:5, but excluding the poly(A) tail at the 3' end of SEQ ID NO:5; and (bb) the nucleotide sequence of the cDNA insert of clone co315_3 deposited under accession number ATCC 98631;
(ii) hybridizing said primer(s) to human genomic DNA in conditions at least as stringent as 4X SSC at 65 degrees C;
(iii) amplifying human DNA sequences; and (iv) isolating the polynucleotide products of step (b)(iii);
wherein at least one isolated polynucleotide comprises a nucleotide sequence selected from the group consisting of:
(v) a nucleotide sequence corresponding to the cDNA sequence of SEQ ID
NO:5, and extending contiguously from a nucleotide sequence corresponding to the 5' end of SEQ ID NO:5 to a nucleotide sequence corresponding to the 3' end of SEQ ID NO:5, but excluding the poly(A) tail at the 3' end of SEQ ID NO:5;
(w) a nucleotide sequence corresponding to the cDNA sequence of SEQ ID
NO:5 from nucleotide 286 to nucleotide 552, and extending contiguously from a nucleotide sequence corresponding to the 5' end of said sequence of SEQ ID
NO:5 from nucleotide 286 to nucleotide 552, to a nucleotide sequence corresponding to the 3' end of said sequence of SEQ ID NO:5 from nucleotide 286 to nucleotide 552;
and (x) a nucleotide sequence corresponding to the cDNA sequence of SEQ ID
NO:5 from nucleotide 475 to nucleotide 552, and extending contiguously from a nucleotide sequence corresponding to the 5' end of said sequence of SEQ ID
NO:5 from nucleotide 475 to nucleotide 552, to a nucleotide sequence corresponding to the 3' end of said sequence of SEQ ID NO:5 from nucleotide 475 to nucleotide 552.
21. 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 1682 to nucleotide 1963;
(c) a polynucleotide comprising the nucleotide sequence of the full-length protein coding sequence of clone ij226_6 deposited under accession number ATCC 98631;
(d) a polynucleotide encoding the full-length protein encoded by the cDNA insert of clone ij226_6 deposited under accession number ATCC 98631;
(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 having biological activity, the fragment comprising eight consecutive amino acids of SEQ ID NO:8;
(g) a polynucleotide which is an allelic variant of a polynucleotide of (a)-(f) above; and (h) a polynucleotide that hybridizes under stringent conditions to any one of the polynucleotides specified in (a)-(f).
22. A protein comprising an amino acid sequence selected from the group consisting of:
(a) the amino acid sequence of SEQ ID NO:8;
(b) fragments of the amino acid sequence of SEQ ID NO:8, each fragment comprising eight consecutive amino acids of SEQ ID NO:8; and (c) the amino acid sequence encoded by the cDNA insert of clone ij226_6 deposited under accession number ATCC 98631;
the protein being substantially free from other mammalian proteins.
23. A process for producing an isolated polynucleotide, wherein the process is 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 NO:7, but excluding the poly(A) tail at the 3' end of SEQ ID NO:7; and (ab) the nucleotide sequence of the cDNA insert of clone ij226_6 deposited under accession number ATCC 98631;
(ii) hybridizing said probe(s) to human genomic DNA in conditions at least as stringent as 4X SSC at 65 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 NO:7, but excluding the poly(A) tail at the 3' end of SEQ ID NO:7; and (bb) the nucleotide sequence of the cDNA insert of clone ij226_6 deposited under accession number ATCC 98631;
(ii) hybridizing said primer(s) to human genomic DNA in conditions at least as stringent as 4X SSC at 65 degrees C;
(iii) amplifying human DNA sequences; and (iv) isolating the polynucleotide products of step (b)(iii);
wherein at least one isolated polynucleotide comprises a nucleotide sequence selected from the group consisting of:

(v) a nucleotide sequence corresponding to the cDNA sequence of SEQ ID
NO:7, and extending contiguously from a nucleotide sequence corresponding to the 5' end of SEQ ID NO:7 to a nucleotide sequence corresponding to the 3' end of SEQ ID NO:7, but excluding the poly(A) tail at the 3' end of SEQ ID NO:7; and (w) a nucleotide sequence corresponding to the cDNA sequence of SEQ ID
NO:7 from nucleotide 1682 to nucleotide 1963, and extending contiguously from a nucleotide sequence corresponding to the 5' end of said sequence of SEQ ID
NO:7 from nucleotide 1682 to nucleotide 1963, to a nucleotide sequence corresponding to the 3' end of said sequence of SEQ ID NO:7 from nucleotide to nucleotide 1963.
24. 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 1137 to nucleotide 1346;
(c) a polynucleotide comprising the nucleotide sequence of the full-length protein coding sequence of clone nf443_1 deposited under accession number ATCC 98631;
(d) a polynucleotide encoding the full-length protein encoded by the cDNA insert of clone nf443_1 deposited under accession number ATCC 98631;
(e) a polynucleotide encoding a protein comprising the amino acid sequence of SEQ ID NO:10;
(f) 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 consecutive amino acids of SEQ ID NO:10;
(g) a polynucleotide which is an allelic variant of a polynucleotide of (a)-(f) above; and (h) a polynucleotide that hybridizes under stringent conditions to any one of the polynucleotides specified in (a)-(f).
25. A protein comprising an amino acid sequence selected from the group consisting of:
(a) the amino acid sequence of SEQ ID NO:10;

(b) fragments of the amino acid sequence of SEQ ID NO:10, each fragment comprising eight consecutive amino acids of SEQ ID NO:10; and (c) the amino acid sequence encoded by the cDNA insert of clone nf443_1 deposited under accession number ATCC 98631;
the protein being substantially free from other mammalian proteins.
26. A process for producing an isolated polynucleotide, wherein the process is 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 NO:9, but excluding the poly(A) tail at the 3' end of SEQ ID NO:9; and (ab) the nucleotide sequence of the cDNA insert of clone nf443_1 deposited under accession number ATCC 98631;
(ii) hybridizing said probe(s) to human genomic DNA in conditions at least as stringent as 4X SSC at 65 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 NO:9, but excluding the poly(A) tail at the 3' end of SEQ ID NO:9; and (bb) the nucleotide sequence of the cDNA insert of clone nf443_1 deposited under accession number ATCC 98631;
(ii) hybridizing said primer(s) to human genomic DNA in conditions at least as stringent as 4X SSC at 65 degrees C;
(iii) amplifying human DNA sequences; and (iv) isolating the polynucleotide products of step (b)(iii);

wherein at least one isolated polynucleotide comprises a nucleotide sequence selected from the group consisting of:
(v) a nucleotide sequence corresponding to the cDNA sequence of SEQ ID
NO:9, and extending contiguously from a nucleotide sequence corresponding to the 5' end of SEQ ID NO:9 to a nucleotide sequence corresponding to the 3' end of SEQ ID NO:9, but excluding the poly(A) tail at the 3' end of SEQ ID NO:9; and (w) a nucleotide sequence corresponding to the cDNA sequence of SEQ ID
NO:9 from nucleotide 1137 to nucleotide 1346, and extending contiguously from a nucleotide sequence corresponding to the 5' end of said sequence of SEQ ID
NO:9 from nucleotide 1137 to nucleotide 1346, to a nucleotide sequence corresponding to the 3' end of said sequence of SEQ ID NO:9 from nucleotide to nucleotide 1346.
27. 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 308 to nucleotide 634;
(c) a polynucleotide comprising the nucleotide sequence of the full-length protein coding sequence of clone nt429_1 deposited under accession number ATCC 98631;
(d) a polynucleotide encoding the full-length protein encoded by the cDNA insert of clone nt429_1 deposited under accession number ATCC 98631;
(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 having biological activity, the fragment comprising eight consecutive amino acids of SEQ ID NO:12;
(g) a polynucleotide which is an allelic variant of a polynucleotide of (a)-(f) above; and (h) a polynucleotide that hybridizes under stringent conditions to any one of the polynucleotides specified in (a)-(f).
28. A protein comprising an amino acid sequence selected from the group consisting of:
(a) the amino acid sequence of SEQ ID NO:12;
(b) the amino acid sequence of SEQ ID NO:12 from amino acid 1 to amino acid 47;
(c) fragments of the amino acid sequence of SEQ ID NO:12, each fragment comprising eight consecutive amino acids of SEQ ID NO:12; and (d) the amino acid sequence encoded by the cDNA insert of clone nt429_1 deposited under accession number ATCC 98631;
the protein being substantially free from other mammalian proteins.
29. A process for producing an isolated polynucleotide, wherein the process is 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 NO: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 nt429_1 deposited under accession number ATCC 98631;
(ii) hybridizing said probe(s) to human genomic DNA in conditions at least as,stringent as 4X SSC at 65 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 NO:11, but excluding the poly(A) tail at the 3' end of SEQ ID NO:11; and (bb) the nucleotide sequence of the cDNA insert of clone nt429_1 deposited under accession number ATCC 98631;

(ii) hybridizing said primer(s) to human genomic DNA in conditions at least as stringent as 4X SSC at 65 degrees C;
(iii) amplifying human DNA sequences; and (iv) isolating the polynucleotide products of step (b)(iii);
wherein at least one isolated polynucleotide comprises a nucleotide sequence selected from the group consisting of:
(v) 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 NO: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;
and (w) a nucleotide sequence corresponding to the cDNA sequence of SEQ ID
NO:11 from nucleotide 308 to nucleotide 634, and extending contiguously from a nucleotide sequence corresponding to the 5' end of said sequence of SEQ ID
NO:11 from nucleotide 308 to nucleotide 634, to a nucleotide sequence corresponding to the 3' end of said sequence of SEQ ID NO:11 from nucleotide 308 to nucleotide 634.
30. 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 104 to nucleotide 652;
(c) a polynucleotide comprising the nucleotide sequence of SEQ ID
NO:13 from nucleotide 377 to nucleotide 652;
(d) a polynucleotide comprising the nucleotide sequence of the full-length protein coding sequence of clone pe503_1 deposited under accession number ATCC 98631;
(e) a polynucleotide encoding the full-length protein encoded by the cDNA insert of clone pe503_1 deposited under accession number ATCC 98631;
(f) a polynucleotide comprising the nucleotide sequence of the mature protein coding sequence of clone pe503_1 deposited under accession number ATCC 98631;

(g) a polynucleotide encoding the mature protein encoded by the cDNA insert of clone pe503_1 deposited under accession number ATCC 98631;
(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 having biological activity, the fragment comprising eight consecutive amino acids of SEQ ID NO:14;
(j) a polynucleotide which is an allelic variant of a polynucleotide of (a)-(g) above; and (k) a polynucleotide that hybridizes under stringent conditions to any one of the polynucleotides specified in (a)-(i).
31. A protein comprising an amino acid sequence selected from the group consisting of:
(a) the amino acid sequence of SEQ ID NO:14;
(b) the amino acid sequence of SEQ ID NO:14 from amino acid 69 to amino acid 125;
(c) fragments of the amino acid sequence of SEQ ID NO:14, each fragment comprising eight consecutive amino acids of SEQ ID NO:14; and (d) the amino acid sequence encoded by the cDNA insert of clone pe503_1 deposited under accession number ATCC 98631;
the protein being substantially free from other mammalian proteins.
32. A process for producing an isolated polynucleotide, wherein the process is 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 NO:13, but excluding the poly(A) tail at the 3' end of SEQ ID NO:13; and (ab) the nucleotide sequence of the cDNA insert of clone pe503_1 deposited under accession number ATCC 98631;

(ii) hybridizing said probe(s) to human genomic DNA in conditions at least as stringent as 4X SSC at 65 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 NO:13, but excluding the poly(A) tail at the 3' end of SEQ ID NO:13; and (bb) the nucleotide sequence of the cDNA insert of clone pe503_1 deposited under accession number ATCC 98631;
(ii) hybridizing said primer(s) to human genomic DNA in conditions at least as stringent as 4X SSC at 65 degrees C;
(iii) amplifying human DNA sequences; and (iv) isolating the polynucleotide products of step (b)(iii);
wherein at least one isolated polynucleotide comprises a nucleotide sequence selected from the group consisting of:
(v) a nucleotide sequence corresponding to the cDNA sequence of SEQ ID
NO:13, and extending contiguously from a nucleotide sequence corresponding to the 5' end of SEQ ID NO:13 to a nucleotide sequence corresponding to the 3' end of SEQ ID NO:13, but excluding the poly(A) tail at the 3' end of SEQ ID NO:13;
(w) a nucleotide sequence corresponding to the cDNA sequence of SEQ ID
NO:13 from nucleotide 104 to nucleotide 652, and extending contiguously from a nucleotide sequence corresponding to the 5' end of said sequence of SEQ ID
NO:13 from nucleotide 104 to nucleotide 652, to a nucleotide sequence corresponding to the 3' end of said sequence of SEQ ID NO:13 from nucleotide 104 to nucleotide 652; and (x) a nucleotide sequence corresponding to the cDNA sequence of SEQ ID
NO:13 from nucleotide 377 to nucleotide 652, and extending contiguously from a nucleotide sequence corresponding to the 5' end of said sequence of SEQ ID
NO:13 from nucleotide 377 to nucleotide 652, to a nucleotide sequence corresponding to the 3' end of said sequence of SEQ ID NO:13 from nucleotide 377 to nucleotide 652.
33. 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 23 to nucleotide 442;
(c) a polynucleotide comprising the nucleotide sequence of SEQ ID
NO:15 from nucleotide 224 to nucleotide 442;
(d) a polynucleotide comprising the nucleotide sequence of the full-length protein coding sequence of clone pe834_6 deposited under accession number ATCC 98631;
(e) a polynucleotide encoding the full-length protein encoded by the cDNA insert of clone pe834_6 deposited under accession number ATCC 98631;
(f) a polynucleotide comprising the nucleotide sequence of the mature protein coding sequence of clone pe834_6 deposited under accession number ATCC 98631;
(g) a polynucleotide encoding the mature protein encoded by the cDNA insert of clone pe834_6 deposited under accession number ATCC 98631;
(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. having biological activity, the fragment comprising eight consecutive amino acids of SEQ ID NO:16;
(j) a polynucleotide which is an allelic variant of a polynucleotide of (a)-(i) above; and (k) a polynucleotide that hybridizes under stringent conditions to any one of the polynucleotides specified in (a)-(i).
34. A protein comprising an amino acid sequence selected from the group consisting of:
(a) the amino acid sequence of SEQ ID NO:16;
(b) fragments of the amino acid sequence of SEQ ID NO:16, each fragment comprising eight consecutive amino acids of SEQ ID NO:16; and (c) the amino acid sequence encoded by the cDNA insert of clone pe834_6 deposited under accession number ATCC 98631;
the protein being substantially free from other mammalian proteins.
35. A process for producing an isolated polynucleotide, wherein the process is 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 N0:15, but excluding the poly(A) tail at the 3' end of SEQ ID NO:15; and (ab) the nucleotide sequence of the cDNA insert of clone pe834_6 deposited under accession number ATCC 98631;
(ii) hybridizing said probe(s) to human genomic DNA in conditions at least as stringent as 4X SSC at 65 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 NO:15, but excluding the poly(A) tail at the 3' end of SEQ ID NO:15; and (bb) the nucleotide sequence of the cDNA insert of clone pe834_6 deposited under accession number ATCC 98631;
(ii) hybridizing said primer(s) to human genomic DNA in conditions at least as stringent as 4X SSC at 65 degrees C;
(iii) amplifying human DNA sequences; and (iv) isolating the polynucleotide products of step (b)(iii);
wherein at least one isolated polynucleotide comprises a nucleotide sequence selected from the group consisting of:

(v) a nucleotide sequence corresponding to the cDNA sequence of SEQ ID
NO:15, and extending contiguously from a nucleotide sequence corresponding to the 5' end of SEQ ID NO:15 to a nucleotide sequence corresponding to the 3' end of SEQ ID NO:15, but excluding the poly(A) tail at the 3' end of SEQ ID NO:15;
and (w) a nucleotide sequence corresponding to the cDNA sequence of SEQ ID
NO:15 from nucleotide 23 to nucleotide 442, and extending contiguously from a nucleotide sequence corresponding to the 5' end of said sequence of SEQ ID
NO:15 from nucleotide 23 to nucleotide 442, to a nucleotide sequence corresponding to the 3' end of said sequence of SEQ ID NO:15 from nucleotide 23 to nucleotide 442.
36. 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 98 to nucleotide 265;
(c) a polynucleotide comprising the nucleotide sequence of SEQ ID
NO:17 from nucleotide 152 to nucleotide 265;
(d) a polynucleotide comprising the nucleotide sequence of the full-length protein coding sequence of clone ya10_1 deposited under accession number ATCC 98631;
(e) a polynucleotide encoding the full-length protein encoded by the cDNA insert of clone ya10_1 deposited under accession number ATCC 98631;
(f) a polynucleotide comprising the nucleotide sequence of the mature protein coding sequence of clone ya10_1 deposited under accession number ATCC
98631;
(g) a polynucleotide encoding the mature protein encoded by the cDNA insert of clone ya10_1 deposited under accession number ATCC 98631;
(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 having biological activity, the fragment comprising eight consecutive amino acids of SEQ ID NO:18;

(j) a polynucleotide which is an allelic variant of a polynucleotide of (a)-(g) above; and (k) a polynucleotide that hybridizes under stringent conditions to any one of the polynucleotides specified in (a)-(i).
37. A protein comprising an amino acid sequence selected from the group consisting of:
(a) the amino acid sequence of SEQ ID NO:18;
(b) fragments of the amino acid sequence of SEQ ID NO:18, each fragment comprising eight consecutive amino acids of SEQ ID NO:18; and (c) the amino acid sequence encoded by the cDNA insert of clone ya10_1 deposited under accession number ATCC 98631;
the protein being substantially free from other mammalian proteins.
38. A process for producing an isolated polynucleotide, wherein the process is 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 NO:17, but excluding the poly(A) tail at the 3' end of SEQ ID NO:17; and (ab) the nucleotide sequence of the cDNA insert of clone ya10_1 deposited under accession number ATCC 98631;
(ii) hybridizing said probe(s) to human genomic DNA in conditions at least as stringent as 4X SSC at 65 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 NO:17, but excluding the poly(A) tail at the 3' end of SEQ ID NO:17; and (bb) the nucleotide sequence of the cDNA insert of clone ya10_1 deposited under accession number ATCC 98631;
(ii) hybridizing said primer(s) to human genomic DNA in conditions at least as stringent as 4X SSC at 65 degrees C;
(iii) amplifying human DNA sequences; and (iv) isolating the polynucleotide products of step (b)(iii);
wherein at least one isolated polynucleotide comprises a nucleotide sequence selected from the group consisting of:
(v) a nucleotide sequence corresponding to the cDNA sequence of SEQ ID
NO:17, and extending contiguously from a nucleotide sequence corresponding to the 5' end of SEQ ID NO:17 to a nucleotide sequence corresponding to the 3' end of SEQ ID NO:17 , but excluding the poly(A) tail at the 3' end of SEQ ID
NO:17;
(w) a nucleotide sequence corresponding to the cDNA sequence of SEQ ID
NO:17 from nucleotide 98 to nucleotide 265, and extending contiguously from a nucleotide sequence corresponding to the 5' end of said sequence of SEQ ID
NO:17 from nucleotide 98 to nucleotide 265, to a nucleotide sequence corresponding to the 3' end of said sequence of SEQ ID NO:17 from nucleotide 98 to nucleotide 265;
and (x) a nucleotide sequence corresponding to the cDNA sequence of SEQ ID
NO:17 from nucleotide 152 to nucleotide 265, and extending contiguously from a nucleotide sequence corresponding to the 5' end of said sequence of SEQ ID
NO:17 from nucleotide 152 to nucleotide 265, to a nucleotide sequence corresponding to the 3' end of said sequence of SEQ ID NO:17 from nucleotide 152 to nucleotide 265.
39. 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 176 to nucleotide 583;

(c) a polynucleotide comprising the nucleotide sequence of the full-length protein coding sequence of clone yb40_1 deposited under accession number ATCC 98631;
(d) a polynucleotide encoding the full-length protein encoded by the cDNA insert of clone yb40_1 deposited under accession number ATCC 98631;
(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 having biological activity, the fragment comprising eight consecutive amino acids of SEQ ID NO:20;
(g) a polynucleotide which is an allelic variant of a polynucleotide of (a)-(f) above; and (h) a polynucleotide that hybridizes under stringent conditions to any one of the polynucleotides specified in (a)-(f).
40. A protein comprising an amino acid sequence selected from the group consisting of:
(a) the amino acid sequence of SEQ ID NO:20;
(b) fragments of the amino acid sequence of SEQ ID NO:20, each fragment comprising eight consecutive amino acids of SEQ ID NO:20; and (c) the amino acid sequence encoded by the cDNA insert of clone yb40_1 deposited under accession number ATCC 98631;
the protein being substantially free from other mammalian proteins.
41. A process for producing an isolated polynucleotide, wherein the process is 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 NO:19, but excluding the poly(A) tail at the 3' end of SEQ ID NO:19; and (ab) the nucleotide sequence of the cDNA insert of clone yb40_1 deposited under accession number ATCC 98631;

(ii) hybridizing said probes) to human genomic DNA in conditions at least as stringent as 4X SSC at 65 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 NO:19, but excluding the poly(A) tail at the 3' end of SEQ ID NO:19; and (bb) the nucleotide sequence of the cDNA insert of clone yb40_1 deposited under accession number ATCC 98631;
(ii) hybridizing said primer(s) to human genomic DNA in conditions at least as stringent as 4X SSC at 65 degrees C;
(iii) amplifying human DNA sequences; and (iv) isolating the polynucleotide products of step (b)(iii);
wherein at least one isolated polynucleotide comprises a nucleotide sequence selected from the group consisting of:
(v) 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 NO:19 to a nucleotide sequence corresponding to the 3' end of SEQ ID NO:19, but excluding the poly(A) tail at the 3' end of SEQ ID NO:19;
and (w) a nucleotide sequence corresponding to the cDNA sequence of SEQ ID
NO:19 from nucleotide 176 to nucleotide 583, and extending contiguously from a nucleotide sequence corresponding to the 5' end of said sequence of SEQ ID
NO:19 from nucleotide 176 to nucleotide 583, to a nucleotide sequence corresponding to the 3' end of said sequence of SEQ ID NO:19 from nucleotide 176 to nucleotide 583.
CA002317392A 1998-01-13 1999-01-11 Secreted proteins and polynucleotides encoding them Abandoned CA2317392A1 (en)

Applications Claiming Priority (5)

Application Number Priority Date Filing Date Title
US7130498P 1998-01-13 1998-01-13
US60/071,304 1998-01-13
US22746299A 1999-01-08 1999-01-08
US09/227,462 1999-01-08
PCT/US1999/000550 WO1999036512A1 (en) 1998-01-13 1999-01-11 Secreted proteins and polynucleotides encoding them

Publications (1)

Publication Number Publication Date
CA2317392A1 true CA2317392A1 (en) 1999-07-22

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EP (1) EP1047772A1 (en)
JP (1) JP2002527038A (en)
AU (1) AU2112099A (en)
CA (1) CA2317392A1 (en)
WO (1) WO1999036512A1 (en)

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JP2002527038A (en) 2002-08-27
WO1999036512A1 (en) 1999-07-22
EP1047772A1 (en) 2000-11-02

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