CA2315245A1 - Secreted proteins and polynucleotides encoding them - Google Patents

Abstract

Novel polynucleotides and the proteins encoded thereby are disclosed.

Description

WO 99/32614 PC"TIUS98IZ7140 SECRETED PROTEINS AND POLYNUCLEOTIDES ENCODING THEM
This application is a continuation-in-part of provisional application Ser. No.
60/068,369, filed December 20,1997, which is incorporated by reference herein.
The present invention provides novel polynucleotides and proteins encoded by such polynucleotides, along with therapeutic, diagnostic and research utilities for these polynucleotides and proteins.
BACKGROUND OF THE INVENTION
Technology aimed at the discovery of protein factors (including e.g., cytokines, such as lymphokines, interferons, CSFs and interleukins) has matured rapidly over the past decade. The now routine hybridization cloning and expression cloning techniques clone novel polynucleotides "directly" in the sense that they rely on information directly related to the discovered protein (i.e., partial DNA/amino acid sequence of the protein in the case of hybridization cloning; activity of the protein in the case of expression 2 0 cloning). More recent "indirect" cloning techniques such as signal sequence cloning, which isolates DNA sequences based on the presence of a now well-recognized secxetory 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 2 5 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 polynucleotides encoding them that the present invention is directed.

In one embodiment, the present invention provides a composition comprising an isolated polynucleotide selected from the group consisting of:
(a) a polynucleotide comprising the nucleotide sequence of SEQ ID
NO:1;
(b) a polynucleotide comprising the nucleotide sequence of SEQ ID
N0:1 from nucleotide 179 to nucleotide 4285;
(c) a polynucleotide comprising the nucleotide sequence of the full-length protein coding sequence of clone am728_60 deposited under accession number ATCC 98621;
(d) a polynucleotide encoding the full-length protein encoded by the cDNA insert of clone am728_60 deposited under accession number ATCC 98621;
(e) a polynucleotide comprising the nucleotide sequence of a mature protein coding sequence of clone am728_60 deposited under accession number ATCC 98621;
(f) a polynucleotide encoding a mature protein encoded by the cDNA
insert of clone am728_60 deposited under accession number ATCC 98621;
(g) a polynucleotide encoding a protein comprising the amino acid sequence of SEQ ID N0:2;
(h) 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;
(i) a polynucleotide which is an allelic variant of a polynucleotide of (a)-(f) above;
2 5 (j) a polynucleotide which encodes a species homologue of the protein of (g) or (h) above ; and (k) a poiynudeotide that hybridizes under stringent conditions to any one of the poiynucleotides specified in (a)-(h).
Preferably, such polynucleotide comprises the nucleotide sequence of SEQ ID

3 0 NO:1 from nucleotide 179 to nucleotide 4285; the nucleotide sequence of the full-length protein coding sequence of clone am728_60 deposited under accession number ATCC
98621; or the nucleotide sequence of a mature protein coding sequence of clone am728_60 deposited under accession number ATCC 98621. In other preferred embodiments, the polynucleotide encodes the full-length or a mature protein encoded by the cDNA
insert of clone am728 60 deposited under accession number ATCC 98621. In further preferred embodiments, the present invention provides a polynudeotide 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 679 to amino acid 688 of SEQ ID N0:2.
Other embodiments provide the gene corresponding to the cDNA sequence of SEQ
ID N0:1.
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 polynudeotide 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; and (ab) the nucleotide sequence of the cDNA insert of clone am728_60 deposited under accession number ATCC 98621;
2 0 (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 polynudeotides detected with the probe(s);
and 2 5 (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; and 3 0 (bb} the nucleotide sequence of the cDNA insert of clone am728_60 deposited under accession number ATCC 98621;
(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 NO: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 179 to nucleotide 4285, and extending contiguously from a nucleotide sequence corresponding to the 5' end of said sequence of SEQ ID N0:1 from nucleotide 179 to nucleotide 4285, to a nucleotide sequence corresponding to the 3' end of said sequence of SEQ ID N0:1 from nucleotide 179 to nucleotide 4285.
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 N0:2;
(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 am728_60 deposited under accession number ATCC 98621;
2 0 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 2 5 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 679 to amino acid 688 of SEQ ID N0:2.
In one embodiment, the present invention provides a composition comprising an isolated polynucleoHde selected from the group consisting of:
3 0 (a) a polynucleotide comprising the nucleotide sequence of SEQ ID
N0:3;
(b) a polynucleotide comprising the nucleotide sequence of SEQ ID
N0:3 from nucleotide 108 to nucleotide 254;

(c) a polynucleotide comprising the nucleotide sequence of SEQ ID
N0:3 from nucleotide 225 to nucleotide 254;
(d) a polynucleotide comprising the nucleotide sequence of the full length protein coding sequence of clone bf3~ 1 deposited under accession number ATCC 98621;
(e) a polynucleotide encoding the full-length protein encoded by the cDNA insert of clone bf377 1 deposited under accession number ATCC 98621;
(f) a polynucleotide comprising the nucleotide sequence of a mature protein coding sequence of clone bf377 1 deposited under accession number ATCC 98621;
(g) a polynucleotide encoding a mature protein encoded by the cDNA
insert of clone bf377 1 deposited under accession number ATCC 98621;
(h) a polynucleotide encoding a protein comprising the amino acid sequence of SEQ ID N0:4;
Z 5 (i) a polynucleotide encoding a protein comprising a fragment of the amino acid sequence of SEQ ID N0:4 having biological activity, the fragment comprising eight consecutive amino acids of SEQ ID N0:4;
(j) a polynucleotide which is an allelic variant of a polynucleotide of (a)-(g) above;
2 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
2 5 N0:3 from nucleotide lOS to nucleotide 254; the nucleotide sequence of SEQ
ID N0:3 from nucleotide 225 to nucleotide 254; the nucleotide sequence of the full-length protein coding sequence of clone bf3~ 1 deposited under accession number ATCC 98621; or the nucleotide sequence of a mature protein coding sequence of clone bf3Tl_1 deposited under accession number ATCC 98621. In other preferred embodiments, the 3 0 polynucleotide encodes the full-length or a mature protein encoded by the cDNA insert of clone bf3~ 1 deposited under accession number ATCC 98621. 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 ands of SEQ ID N0:4, or a polynucleotide encoding a protein comprising a fragment of the amino acid sequence of SEQ ID N0:4 having biological activity, the fragment comprising the amino acid sequence from amino acid 19 to amino acid 28 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:
(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 3' end of SEQ ID N0:3; and (ab) the nucleotide sequence of the cDNA insert of clone bf377_1 deposited under accession number ATCC 98621;
(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 2 0 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 2 5 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 bf377_1 deposited under accession number ATCC 98621;
3 0 (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 108 to nucleotide 254, and extending contiguously from a nucleotide sequence corresponding to the 5' end of said sequence of SEQ ID N0:3 from nucleotide 108 to nucleotide 254, to a nucleotide sequence corresponding to the 3' end of said sequence of SEQ ID N0:3 from nucleotide 108 to nucleotide 254. 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 225 to nucleotide 254, and extending contiguously from a nucleotide sequence corresponding to the 5' end of said sequence of SEQ ID
N0:3 from nucleotide 225 to nucleotide 254, to a nucleotide sequence corresponding to the 3' end of said sequence of SEQ ID N0:3 from nucleotide 225 to nucleotide 254.
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:
2 0 (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 (c) the amino acid sequence encoded by the cDNA insert of clone bf3~ 1 deposited under accession number ATCC 98621;
2 5 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 3 0 ands 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 and 19 to amino acid 28 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 426 to nucleotide 569;
(c) a polynucleotide comprising the nucleotide sequence of SEQ ID
N0:5 from nucleotide 546 to nucleotide 569;
(d) a polynucleotide comprising the nucleotide sequence of the full-length protein coding sequence of clone cw354_l deposited under accession number ATCC 98621;
(e) a polynucleotide encoding the full-length protein encoded by the cDNA insert of clone cw354_1 deposited under accession number ATCC 98621;
(f] a polynucleotide comprising the nucleotide sequence of a mature protein coding sequence of clone cw354_1 deposited under accession number ATCC 98621;
(g) a polynucleotide encoding a mature protein encoded by the cDNA
insert of clone cw354_1 deposited under accession number ATCC 98621;
(h) a polynucleotide encoding a protein comprising the amino acid sequence of SEQ ID N0:6;
(i) a polynucleotide encoding a protein comprising a fragment of the 2 0 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 polynucleotide which is an allelic variant of a polynucleotide of (a)-(g) above;
(k) a polynucleotide which encodes a species homologue of the protein 2 5 of (h) or (i) above ; 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
N0:5 from nucleotide 426 to nucleotide 569; the nucleotide sequence of SEQ ID
N0:5 from 3 0 nucleotide 546 to nucleotide 569; the nucleotide sequence of the full-length protein coding sequence of clone cw354_1 deposited under accession number ATCC 98621; or the nucleotide sequence of a mature protein coding sequence of clone cw354_1 deposited under accession number ATCC 98621. In other preferred embodiments, the polynudeotide encodes the full-length or a mature protein encoded by the cDNA
insert of clone cw354_1 deposited under accession number ATCC 98621. 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 polynucleotide encoding 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 19 to amino acid 28 of SEQ ID N0:6.
Other embodiments provide the gene corresponding to the cDNA sequence of SEQ
ID NO:S.
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 N0:5, 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 2 0 cw354_1 deposited under accession number ATCC 98621;
(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 probes};
2 5 and (b) a process comprising the steps of:
(i) preparing one or more polynucleotide primers that hybridize in 6X SSC at 65 degrees C to a nucleotide sequence selected from the group consisting of:
3 0 (ba) SEQ ID 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 cw354-1 deposited under accession number ATCC 98621;

(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 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 kom nucleotide 426 to nucleotide 569, and extending contiguously from a nucleotide sequence corresponding to the 5' end of said sequence of SEQ ID N0:5 from nucleotide 426 to nucleotide 569, to a nucleotide sequence corresponding to the 3' end of said sequence of SEQ ID N0:5 from nucleotide 426 to 1 S nucleotide 569. Also preferably the polynucleotide isolated according to the above process comprises a nucleotide sequence corresponding to the cDNA sequence of SEQ ID
N0:5 kom nucleotide 546 to nucleotide 569, and extending contiguously from a nucleotide sequence corresponding to the 5' end of said sequence of SEQ ID
N0:5 from nucleotide 546 to nucleotide 569, to a nucleotide sequence corresponding to the 3' end of 2 0 said sequence of SEQ ID N0:5 kom nucleotide 546 to nucleotide 569.
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;
2 5 (b) fragments of the amino acid sequence of SEQ ID N0:6, each kagment comprising eight consecutive amino acids of SEQ ID N0:6; and (c) the amino acid sequence encoded by the cDNA insert of clone cw354_1 deposited under accession number ATCC 9$621;
the protein being substantially free from other mammalian proteins. Preferably such 3 0 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 kagment of the amino acid sequence of SEQ ID N0:6 having biological activity, the fragment comprising the amino acid sequence from amino acid 19 to amino acid 28 of SEQ ID N0:6.
In one embodiment, the present invention provides a composition comprising an isolated polynucleotide selected from the group consisting of:
(a) a polynucleotide comprising the nucleotide sequence of SEQ ID
N0:7;
(b) a polynucleotide comprising the nucleotide sequence of SEQ ID
N0:7 from nucleotide 151 to nucleotide 891;
(c) a polynucleotide comprising the nucleotide sequence of SEQ ID
N0:7 from nucleotide 595 to nucleotide 891;
(d) a polynucleotide comprising the nucleotide sequence of the full-length protein coding sequence of clone nm134 4 deposited under accession number ATCC 98621;
(e} a polynucleotide encoding the full-length protein encoded by the cDNA insert of clone nm134 4 deposited under accession number ATCC 98621;
(f) a polynucleotide comprising the nucleotide sequence of a mature protein coding sequence of clone nm134 4 deposited under accession number ATCC 98621;
(g) a polynucleotide encoding a mature protein encoded by the cDNA
2 0 insert of clone nm134 4 deposited under accession number ATCC 98621;
(h) a polynucleotide encoding a protein comprising the amino acid sequence of SEQ ID N0:8;
(i) a polynucleotide encoding a protein comprising a fragment of the amino acid sequence of SEQ ID N0:8 having biological activity, the fragment 2 5 rnmprising eight consecutive amino acids of SEQ ID N0:8;
(j) a polynucleotide which is an allelic variant of a polynucleotide of (ar(g) above;
(k) a polynucleotide which encodes a species homologue of the protein of (h) or (i) above ; and 3 0 (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
N0:7 from nucleotide 151 to nucleotide 891; the nucleotide sequence of SEQ ID
N0:7 from nucleotide 595 to nucleotide 891; the nucleotide sequence of the full-length protein coding a sequence of clone -nm134 4 deposited under accession number ATCC 98621; or the nucleotide sequence of a mature protein coding sequence of clone nm134 4 deposited under accession number ATCC 98621. In other preferred embodiments, the polynucleotide encodes the full-length or a mature protein encoded by the cDNA
insert of clone nm134 4 deposited under accession number ATCC 98621. In yet other preferred embodiments, the present invention provides a polynucleotide encoding a protein comprising the amino acid sequence of SEQ ID N0:8 from amino acid 104 to amino acid 163. 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 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 118 to amino acid 127 of SEQ ID N0:8.
Other embodiments provide the gene corresponding to the cDNA sequence of SEQ
ID N0:7.
Further embodiments of the invention provide isolated polynudeotides produced according to a process selected from the group consisting of:
(a) a process comprising the steps of:
2 0 (i) preparing one or more polynucleotide probes that hybridize in 6X SSC at 65 degrees C to a nucleotide sequence selected from the group consisting of:
(aa) SEQ ID N0:7, but excluding the poly(A) tail at the 3' end of SEQ ID N0:7; and 2 5 (ab} the nucleotide sequence of the cDNA insert of clone run134 4 deposited under accession number ATCC 98621;
(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 3 0 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 nm134 4 deposited under accession number ATCC 98621;
(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 N0:7, and extending contiguously from a nucleotide sequence corresponding to the 5' end of SEQ ID
N0:7 to a nucleotide sequence corresponding to the 3' end of SEQ ID N0:7 , but excluding the poly(A) tail at the 3' end of SEQ ID N0:7. Also preferably the polynucleotide isolated according to the above process comprises a nucleotide sequence corresponding to the cDNA sequence of SEQ ID N0:7 from nucleotide 151 to nucleotide 891, and extending contiguously from a nucleotide sequence corresponding to the 5' end of said sequence of 2 0 SEQ ID N0:7 from nucleotide 151 to nucleotide 891, to a nucleotide sequence corresponding to the 3' end of said sequence of SEQ ID N0:7 from nucleotide 151 to nucleotide 891. Also preferably the polynucleotide isolated according to the above process comprises a nucleotide sequence corresponding to the cDNA sequence of SEQ ID
N0:7 from nucleotide 595 to nucleotide 891, and extending contiguously from a 2 5 nucleotide sequence corresponding to the 5' end of said sequence of SEQ ID
N0:7 from nucleotide 595 to nucleotide 891, to a nucleotide sequence corresponding to the 3' end of said sequence of SEQ ID N0:7 from nucleotide 595 to nucleotide 891.
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 N0:8;
(b) the amino and sequence of SEQ ID N0:8 from amino acid 104 to amino acid 163;

WO 99!32614 PGT/US98J27140 (c) fragments of the amino acid sequence of SEQ ID N0:8, each fragment comprising eight consecutive amino acids of SEQ ID N0:8; and (d) the amino acid sequence encoded by the cDNA insert of clone nm134 4 deposited under accession number ATCC 98621;
the protein being substantially free from other mammalian proteins. Preferably such protein comprises the amino acid sequence of SEQ ID N0:8 or the amino acid sequence of SEQ ID N0:8 from amino acid 104 to amino acid 163. 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 118 to amino acid 127 of SEQ ID N0:8.
In one embodiment, the present invention provides a composition comprising an isolated polynucleotide selected from the group consisting of:
(a) a polynucleotide comprising the nucleotide sequence. of SEQ ID
N0:9;
(b} a polynucleotide comprising the nucleotide sequence of SEQ ID
N0:9 from nucleotide 1909 to nucleotide 2127;
2 0 (c) a polynucleotide comprising the nucleotide sequence of SEQ ID
N0:9 from nucleotide 2074 to nucleotide 2127;
(d) a polynucleotide comprising the nucleotide sequence of the full-length protein coding sequence of clone ybll 1 deposited under accession number ATCC 98621;
2 5 (e) a polynucleotide encoding the full-length protein encoded by the cDNA insert of clone ybll_1 deposited under accession number ATCC 98621;
(f) a polynucleotide comprising the nucleotide sequence of a mature protein coding sequence of clone ybl l 1 deposited under accession number ATCC
98621;
3 0 (g) a polynucleotide encoding a mature protein encoded by the cDNA
insert of clone ybll_1 deposited under accession number ATCC 98621;
(h) a polynucleotide encoding a protein comprising the amino acid sequence of SEQ ID N0:10;

WO 99/32614 PGT/US9$/27140 (i) 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;
(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
N0:9 from nucleotide 1909 to nucleotide 2127; the nucleotide sequence of SEQ
ID N0:9 from nucleotide 2074 to nucleotide 2127; the nucleotide sequence of the full-length protein coding sequence of clone ybll 1 deposited under accession number ATCC
98621;
or the nucleotide sequence of a mature protein coding sequence of clone ybll 1 deposited under accession number ATCC 98621. In other preferred embodiments, the polynucleotide encodes the full-length or a mature protein encoded by the cDNA
insert of clone ybll 1 deposited under accession number ATCC 98621. In further preferred embodiments, the present invention provides a polynucleotide encoding a protein comprising a fragment of the amino acid sequence of SEQ ID N0:10 having biological 2 0 activity, the fragment preferably comprising eight (more preferably twenty, most preferably thirty) consecutive amino acids of SEQ ID NO: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 31 to amino acid 40 of SEQ ID N0:10.
2 5 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:
3 0 (i) preparing one or more polynucleotide probes that hybridize in 6X SSC at 65 degrees C to a nucleotide sequ~ce selected from the group consisting of:
(aa) SEQ ID N0:9, but excluding the poly{A) tail at the 3' end of SEQ ID N0:9; and WO 99!31614 PCTNS98/x7140 (ab) the nucleotide sequence of the cDNA insert of clone ybll_1 deposited under accession number ATCC 98621;
(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 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 ybl l 1 deposited under accession number ATCC 98621;
(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 N0:9, and extending contiguously from a nucleotide sequence corresponding to the 5' end of SEQ ID
N0:9 to a nucleotide sequence corresponding to the 3' end of SEQ ID N0:9 , but excluding the poly(A) tail at the 3' end of SEQ ID N0:9. Also preferably the polynucleotide isolated 2 5 according to the above process comprises a nucleotide sequence corresponding to the cDNA sequence of SEQ ID N0:9 from nucleotide 1909 to nucleotide 2127, and extending contiguously from a nucleotide sequence corresponding to the 5' end of said sequence of SEQ ID N0:9 from nucleotide 1909 to nucleotide 2127, to a nucleotide sequence corresponding to the 3' end of said sequence of SEQ ID N0:9 from nucleotide 1909 to 3 0 nucleotide 2127. 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 2074 to nucleotide 2127, and extending contiguously from a nucleotide sequence corresponding to the 5' end of said sequence of SEQ ID
N0:9 from nucleotide 2074 to nucleotide 2127, to a nucleotide sequence corresponding to the 3' end of said sequence of SEQ ID N0:9 from nucleotide 2x74 to nucleotide 2127.
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 N0:10;
(b) fragments of the amino acid sequence of SEQ ID NO:IO, 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 ybl l 1 deposited under accession number ATCC 98621;
the protein being substantially free from other mammalian proteins. Preferably such protein comprises the amino acid sequence of SEQ ID N0:10. 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 31 to amino acid 40 of SEQ ID N0:10.
In one embodiment, the present invention provides a composition comprising an 2 0 isolated polynucleotide selected from the group consisting of:
(a) a polynucleotide comprising the nucleotide sequence of SEQ ID
NO:11;
(b) a polynucleotide comprising the nucleotide sequence of SEQ ID
N0:11 from nucleotide 1077 to nucleotide 1733;
(c) a polynucleotide comprising the nucleotide sequence of SEQ ID
N0:11 from nucleotide 1158 to nucleotide 1733;
(d) a polynucleotide comprising the nucleotide sequence of the full-length protein coding sequence of clone y~ 1 deposited under accession number ATCC 98621;
3 0 (e) a polynucleotide encoding the full-length protein encoded by the cDNA insert of clone y~ 1 deposited under accession number ATCC 98621;
(f) a polynucleotide comprising the nucleotide sequence of a mature protein coding sequence of done yc2_1 deposited under accession number ATCC
98621;

WO 99132614 PCT/US98l27140 (g) a polynucleodde encoding a mature protein encoded by the cDNA
insert of clone y~ 1 deposited under accession number ATCC 98621;
(h) a polynucleotide encoding a protein comprising the amino acid sequence of SEQ ID N0:12;
(i) a polynucleotide encoding a protein comprising a fragment of the amino acid sequence of SEQ ID N0:12 having biological activity, the fragment comprising eight consecutive amino acids of SEQ ID N0:12;
(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
NO:11 from nucleotide 1077 to nucleotide 1733; the nucleotide sequence of SEQ
ID NO:11 from nucleotide 1158 to nucleotide 1733; the nucleotide sequence of the full-length protein coding sequence of clone y~ 1 deposited under accession number ATCC
98621;
or the nucleotide sequence of a mature protein coding sequence of clone y~ 1 deposited under accession number ATCC 98621. In other preferred embodiments, the 2 0 polynucleotide encodes the full-length or a mature protein encoded by the cDNA insert of clone y~ 1 deposited under accession number ATCC 98621. 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 2 5 preferably thirty) consecutive amino ands 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 104 to amino acid 113 of SEQ ID N0:12.
Other embodiments provide the gene corresponding to the cDNA sequence of SEQ
3 0 ID NO:11.
Further embodiments of the invention provide isolated polynucleotides produced according to a process selected from the group consisting of:
(a) a process comprising the steps of:

(i) preparing one or more polynudeotide 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 N0:11; and (ab) the nucleotide sequence of the cDNA insert of done yc2_1 deposited under accession number ATCC 98621;
(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 polynudeotides detected with the probe(s);
and (b) a process comprising the steps of:
(i) preparing one or more polynudeotide 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 2 0 y~ 1 deposited under accession number ATCC 98621;
(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 polynudeotide products of step (b)(iii).
2 5 Preferably the polynucleotide isolated according to the above process comprises a nucleotide sequence corresponding to the cDNA sequence of SEQ ID NO:11, and extending contiguously from a nucleotide sequence corresponding to the 5' end of SEQ
ID N0:11 to a nucleotide sequence corresponding to the 3' end of SEQ ID NO:11, but excluding the poly(A) tail at the 3' end of SEQ ID N0:11. Also preferably the 3 0 polynucleotide isolated according to the above process comprises a nucleotide sequence rnrresponding to the cDNA sequence of SEQ ID NO:11 from nucleotide 1077 to nucleotide 1733, and extending contiguously from a nucleotide sequence corresponding to the 5' end of said sequence of SEQ ID N0:11 from nucleotide 1077 to nucleotide 1733, to a nucleotide sequence corresponding to the 3' end of said sequence of SEQ ID N0:11 from nucleotide 1077 to nucleotide 1733. Also preferably the polynucleotide isolated according to the above process comprises a nucleotide sequence corresponding to the cDNA
sequence of SEQ ID N0:11 from nucleotide 1158 to nucleotide 1733, and extending contiguously from a nucleotide sequence corresponding to the 5' end of said sequence of SEQ ID
N0:11 from nucleotide 1158 to nucleotide 1733, to a nucleotide sequence corresponding to the 3' end of said sequence of SEQ ID NO:11 from nucleotide 1158 to nucleotide 1733.
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) fragments of the amino acid sequence of SEQ ID N0:12, each fragment comprising eight consecutive amino acids of SEQ ID N0:12; and {c) the amino acid sequence encoded by the cDNA insert of clone y~ 1 deposited under accession number ATCC 98621;
the protein being substantially free from other mammalian proteins. Preferably such protein comprises the amino acid sequence of SEQ ID N0:12. In further preferred embodiments, the present invention provides a protein comprising a fragment of the amino acid sequence of SEQ ID N0:12 having biological activity, the fragment preferably comprising eight (more preferably twenty, most preferably thirty) consecutive amino 2 0 ands 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 104 to amino acid 113 of SEQ ID N0:12.
In certain preferred embodiments, the polynucleotide is operably linked to an expression control sequence. The invention also provides a host cell, including bacterial, 2 5 yeast, insect and mammalian cells, transformed with such polynudeotide compositions.
Also provided by the present invention are organisms that have enhanced, reduced, or modified expression of the genes) corresponding to the polynucleotide sequences disclosed herein.
Processes are also provided for producing a protein, which comprise:
3 0 (a) growing a culture of the host cell transformed with such polynucleotide compositions in a suitable culture medium; and (b) purifying the protein from the culture.
The protein produced according to such methods is also provided by the present invention.

Protein compositions of the present invention may further comprise a pharmaceutically acceptable Garner. 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 DRAWINGS
Figures lA and 1B are schematic representations of the pED6 and pNOTs vectors, respectively, used for deposit of clones disclosed herein.
D~~AILED 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 endoplasmic reticulum.
Clone "am728 60"
A polynucleotide of the present invention has been identified as clone "am728_60".
am728_60 was isolated from a human fetal kidney (293 cell line} cDNA library using methods which are selective for cDNAs encoding secreted proteins (see U.S.
Pat. No.
5,536,637), or was identified as encoding a secreted or transmembrane protein on the basis of computer analysis of the amino acid sequence of the encoded protein.
am728_60 is a full-length clone, including the entire coding sequence of a secreted protein (also referred to herein as "am728_60 protein").
The nucleotide sequence of am728_60 as presently determined is reported in SEQ
ID NO:1. What applicants presently believe to be the proper reading frame and the predicted amino acid sequence of the am728_60 protein corresponding to the foregoing nucleotide sequence is reported in SEQ ID N0:2.
The EcoRI/NotI restriction fragment obtainable from the deposit containing clone am728_60 should be approximately 4333 bp.
The nucleotide sequence disclosed herein for am728_60 was searched against the GenBank and GeneSeq nucleotide sequence databases using BLASTN/BLASTX and FASTA search protocols. am728_60 demonstrated at least some similarity with sequences identified as AA446039 (zw66a08.r1 Scares testis NHT Homo Sapiens cDNA clone 5') and U73682 (Human meningioma-expressed antigen 11 (MEA11) mRNA, partial cds).
The predicted amino acid sequence disclosed herein for am728_60 was searched against the GenPept and GeneSeq amino acid sequence databases using the BLASTX search protocol. The predicted am728_60 protein demonstrated at least some similarity to 2 0 sequences identified as U67884 (melanoma inhibitory activity/condrocyte-derived retinoic acid sensitive protein homolog [Rattus norvegicus]), U73682 (meningioma-expressed antigen 11 [Homo sapiens]), U94780 (MEA6 [Homo Sapiens]), and X84707 (melanoma growth regulatory protein [Homo sapiens]). Based upon sequence similarity, am728_60 proteins and each similar protein or peptide may share at least some activity.
The 2 5 TopPredII computer program predicts three potential transmembrane domains within the am728_60 protein sequence, centered around amino acids 300, 370, and 670 of SEQ ID
N0:2, respectively.
When expressed in COS cells, am728_60 protein was detected in a membrane fraction from these cells as a band migrating at approximately 200 kD on a denaturing 3 0 SDS polyacrylamide gel.
done "bf377 1"
A polynucleotide of the present invention has been identified as clone "bf377 1".
bf377 1 was isolated from a human fetal 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. bf377 1 is a full-length clone, including the entire coding sequence of a secreted protein (also referred to herein as 'bf377_1 protein").
The nucleotide sequence of bf377 1 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 bf377 1 protein corresponding to the foregoing nucleotide sequence is reported in SEQ ID N0:4.
Amino acids 27 to 39 of SEQ ID N0:4 are a predicted leader/signal sequence, with the predicted mature amino acid sequence beginning at amino acid 40. 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 bf377 1 protein.
The EcoRI/NotI restriction fragment obtainable from the deposit containing clone bf377 1 should be approximately 450 bp.
The nucleotide sequence disclosed herein for bf377_1 was searched against the GenBank and GeneSeq nucleotide sequence databases using BT ASTN/BLASTX and FASTA search protocols. bf377_1 demonstrated at least some similarity with sequences 2 0 identified as AA559859 (n148c05.s1 NCI CGAP Pr4 Homo Sapiens cDNA clone IMAGE
1043912), AA657838 (nu08bll.sl NCI_CGAP_Pr2 Homo sapiens cDNA clone IMAGE:1207389 similar to gb:M15990 PROTO-ONCOGENE TYROSINE-PROTEIN
ICINASE YES (HUMAN)), and 849353 (yg67e07.s1 Homo Sapiens cDNA clone 38126 3' similar to contains MER22 repetitive element). Based upon sequence similarity, bf377 1 proteins and each similar protein or peptide may share at least some activity.
Clone "cw3 1"
A polynucleotide of the present invention has been identified as clone "cw354 1".
cw354_1 was isolated from a human fetal brain cDNA library using methods which are 3 0 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. cw354_1 is a full-length clone, including the entire coding sequence of a secreted protein (also referred to herein as "cw354_1 protein").

WO 99/32614 PCTIUS9812'1140 The nucleotide sequence of cw354_1 as presently determined is reported in SEQ
ID N0:5, and includes a poly(A) tail. What applicants presently believe to be the proper reading frame and the predicted amino acid sequence of the cw354_1 protein corresponding to the foregoing nucleotide sequence is reported in SEQ ID N0:6.
Amino acids 28 to 40 of SEQ ID N0:6 are a predicted leader/signal sequence, with the predicted mature amino acid sequence beginning at amino acid 41. 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 cw354_1 protein.
The EcoRI/Notl restriction fragment obtainable from the deposit containing clone cw354_1 should be approximately 1350 bp.
The nucleotide sequence disclosed herein for cw354_i was searched against the GenBank and GeneSeq nucleotide sequence databases using BLASTN/BLASTX and FASTA search protocols. cw354_1 demonstrated at least some similarity with sequences identified as D58859 (Human placenta cDNA 5'-end GEN-514B03), H07863 (y186b05.s1 Homo sapiens cDNA clone 45017 3'), N32178 (yy25b09.s1 Homo sapiens cDNA clone 272249 3'), 881953 (yi98e11.r1 Homo Sapiens cDNA clone 147308 5'), and W84437 (zd89d06.s1 Soares fetal heart NbHHI9W Homo sapiens cDNA clone 356651 3'). The predicted amino acid sequence disclosed herein for cw354_1 was searched against the 2 0 GenPept and GeneSeq amino acid sequence databases using the BLASTX search protocol.
The predicted cw354_1 protein demonstrated at least some similarity to sequences identified as U39726 (adenosinetriphosphatase (Mycoplasma gerutalium]). Based upon sequence similarity, cw354_1 proteins and each similar protein or peptide may share at least some activity.
Clone "nm134 4"
A polynucleotide of the present invention has been identified as clone "nm134_4".
nm134 4 was isolated from a human adult blood (erythroleukemia TF-1) cDNA
library using methods which are selective for cDNAs encoding secreted proteins (see U.S. Pat.
3 0 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.
nm134_4 is a full-length clone, including the entire coding sequence of a secreted protein (also referred to herein as "nm134_4 protein").

WO 99/32614 PCT/US98/~7140 The nucleotide sequence of nm134_4 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 nm134 4 protein corresponding to the foregoing nucleotide sequence is reported in SEQ ID N0:8.
Amino acids 136 to 148 of SEQ ID N0:8 are a predicted leader/signal sequence, with the predicted mature amino acid sequence beginning at amino acid 149. 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 nm134 4 protein.
The EcoRI/NotI restriction fragment obtainable from the deposit containing clone nm134 4 should be approximately 1500 bp.
The nucleotide sequence disclosed herein for nm134_4 was searched against the GenBank and GeneSeq nucleotide sequence databases using BLASTN/BLASTX and FASTA search protocols. nm134_4 demonstrated at least some similarity with sequences identified as AA205020 (zq72a12x1 Stratagene neuroepithelium (#937231) Homo sapiens cDNA clone 647134 5'), AA205286 (zq72a12.s1 Stratagene neuroepithelium (#937231) Homo sapiens cDNA clone 647134 3'), AA261864 (zs18h05.r1 Soares NbHTGBC Homo sapiens cDNA clone 685593 5'), and H63680 (yr55d03.r1 Homo sapiens cDNA clone 209189 5'). Based upon sequence similarity, nm134 4 proteins and each similar protein 2 0 or peptide may share at least some activity. The TopPredII computer program predicts five potential transmembrane domains within the nm134_4 protein sequence centered around amino acids 108,132,170,195, and 226 of SEQ ID N0:8, respectively.
Clone "yrbll 1"
2 5 A polynucleotide of the present invention has been identified as clone "ybl l 1".
ybll 1 was isolated from a human fetal brain cDNA library and was identified as encoding a secreted or transmembrane protein on the basis of computer analysis of the amino aad sequence of the encoded protein. ybll 1 is a full-length clone, including the entire coding sequence of a secreted protein (also referred to herein as "ybl l 1 protein").
3 0 The nucleotide sequence of ybI l_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 ybll_1 protein corresponding to the foregoing nucleotide sequence is reported in SEQ ID
NO:10. Amino acids 43 to 55 of SEQ ID NO:10 are a predicted leader/signal sequence, with the predicted mature amino acid sequence beginning at amino acid 56. 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 ybll 1 protein.
The EcoRI/NotI restriction fragment obtainable from the deposit containing clone ybl2_1 should be approximately 2800 bp.
The nucleotide sequence disclosed herein for ybll 1 was searched against the GenBank and GeneSeq nucleotide sequence databases u&ing BLASTN/BLASTX and FASTA search protocols. ybll 1 demonstrated at least some similarity with sequences identified as 855695 (yg88f12.s1 Homo sapiens cDNA done 40397 3') and 885100 (yo43b05.s1 Homo sapiens cDNA clone 180657 3'). Based upon sequence similarity, ybll_1 proteins and each similar protein or peptide may share at least some activity.
Clone "~~ ~' A polynudeotide of the present invention has been identified as clone "yc2_1".
yc2_1 was isolated from a human fetal kidney (293 cell line) 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. yc2_1 is a full-length clone, including the entire coding sequence of a secreted protein (also referred to herein as 2 0 "yc2_1 protein").
The nucleotide sequence of y~ 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 reading frame and the predicted amino acid sequence of the y~ 1 protein corresponding to the foregoing nucleotide sequence is reported in SEQ ID N0:12. Amino acids 15 to 27 2 5 of SEQ ID N0:12 are a predicted leader/signal sequence, with the predicted mature amino acid sequence beginning at amino acid 28. 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 y~ 1 protein.
3 0 The EcoRI/NotI restriction fragment obtainable from the deposit containing clone y~ 1 should be approximately 2900 bp.
The nucleotide sequence disclosed herein for y~ 1 was searched against the GenBank and GeneSeq nucleotide sequence databases using BLASTN/BLASTX and FASTA search protocols. yc2_i demonstrated at least some similarity with sequences identified as AA618531 (np38a03.s1 NCI CGAP_Lul Homo Sapiens cDNA clone IMAGE:1118572 similar to contains Alu repetitive element) and AA626937 (af84h07.s1 Soares testis NHT Homo sapiens cDNA clone 1048765 3'). Based upon sequence similarity, y~ 1 proteins and each similar protein or peptide may share at least some activity. The nucleotide sequence of y~ 1 indicates that it may contain one or more Alu repetitive elements.
Clones am728_60, bf377 1, cw354_l, nm134_4, ybl l 1, and yc2 1 were deposited on December 19, 1997 with the American Type Culture Collection (10801 University Boulevard, Manassas, Virginia 20110-2209 U.S.A.) as an original deposit under the Budapest Treaty and were given the accession number ATCC 98621, from which each clone comprising a particular polynucleotide is obtainable. All restrictions on the availability to the public of the deposited material will be irrevocably removed upon the granting of the patent, except for the requirements specified in 37 C.F.R. ~
1.808(b), and the term of the deposit will comply with 37 C.F.R. ~ 1.806.
Each clone has been transfected into separate bacterial cells (E. coli) in this rnmposite 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 2 0 appropriate fragment for such clone. Each clone was deposited in either the pED6 or pNOTs vector depicted in Figures 1A and 1B, respectively. The pED6dpc2 vector ("pED6") was derived from pED6dpc1 by insertion of a new polylinker to facilitate cDNA cloning (Kaufman et al.,1991, Nucleic Acids Res.19: 4485-4490); the pNOTs vector was derived from pMT2 (Kaufman et al.,1989, Mol. Cell. Biol. 9: 946-958) by deletion of 2 5 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 3 0 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.
Cl-ne Pro SPd ~ n a SEQ ID N0:13 ~"'~-1 SEQ ID N0:14 nm134_4 SEQ ID N0:15 ybll 1 SEQ ID NO:I6 Y~ 1 SEQ ID N0:17 In the sequences listed above which include an N at position 2, that position is occupied in preferred probes/primers by a biotinylated phosphoaramidite residue rather than a nucleotide (such as, for example, that produced by use of biotin phosphoramidite (1-dimethoxytrityloxy-2-(N-biotinyl-4-aminobutyl)-propyl-3-O-(2-cyanoethyl)-(N,N-diisopropyl)-phosphoramadite) (Glen Research, cat. no.10-1953)).
The design of the oligonucleotide probe should preferably follow these 2 0 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).
2 5 The oligonucleotide should preferably be labeled with ~y-~P 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 3 0 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 100 111 of the stock used to inoculate a sterile culture flask containing 25 aril of sterile L-broth containing ampicillin at 100 Ilg/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 ug/m1 and agar at 1.5% in a 150 mm petri dish when grown overnight at 37°C. Other known methods of obtaining distinct, well-separated colonies can also be employed.
Standard colony hybridization procedures should then be used to transfer the colonies to nitrocellulose filters and lyre, 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 Ilg/ml of yeast RNA, and 10 mM EDTA
(approximately 10 mL per 150 mm filter). Preferably, the probe is then added to the hybridization mix at a concentration greater than or equal to 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 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 2 0 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 2 5 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 ~Q, 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 carrier molecules such as 3 0 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 polynucleotide (preferably those deposited with ATCC) in a suitable mammalian cell or other host cell. The sequences) of the mature forms) of the protein may also be determinable from the amino acid sequence of the full-length form.
The present invention also provides genes corresponding to the polynucleotide sequences disclosed herein. "Corresponding genes" are the regions of the genome that are transcribed to produce the mRNAs from which cDNA polynucleotide sequences are derived and may include contiguous regions of the genome necessary for the regulated expression of such genes. Corresponding genes may therefore include but are not limited to coding sequences, 5' and 3' untranslated regions, alternatively spliced exons, introns, promoters, enhancers, and silencer or suppressor elements. The corresponding genes can be isolated in accordance with known methods using the sequence information disclosed herein. Such methods include the preparation of probes or primers from the disclosed sequence information for identification and/or amplification of genes in appropriate 2 0 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 2 5 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 3 0 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.ruh.gov/UniGene/, in order to WO 99132614 PCTIUS98r17140 identify "UniGene clusters" of overlapping sequences. Many of the "UruGene clusters"
so identified will already have been mapped to particular chromosomal sites.
Organisms that have enhanced, reduced, or modified expression of the genes) corresponding to the polynucleotide sequences disclosed herein are provided.
The desired change in gene expression can be achieved through the use of antisense polynucleotides or ribozymes that bind and/or cleave the mRNA transcribed from the gene (Albert and Morris,1994, Trends Pharmacol. Sci.15(7): 250-254; lravarosky 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 herein, preferably produced by transformation of cells with genetic constructs that are stably maintained within the transformed cells and their progeny, are provided.
Transgenic animals that have modified genetic control regions that increase or reduce gene expression levels, or that change temporal or spatial patterns of gene expression, are also provided (see European Patent No. 0 649 464 Bl, incorporated by reference herein).
In addition, organisms are provided in which the genes) corresponding to the polynucleotide sequences disclosed herein have been partially or completely inactivated, through insertion of extraneous sequences into the corresponding genes) or through deletion of all or part of the corresponding gene(s). Partial or complete gene inactivation 2 0 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(I6): 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, 2 5 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 3 0 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 WO 99/32614 PCT/US98/2'f140 the protein is fully secreted from the cell in which it is expressed. The intracellular and transmembrane domains of proteins of the invention can be identified in accordance with known techniques for determination of such domains from sequence information.
For example, the TopPredII computer program can be used to predict the location of transmembrane domains in an amino acid sequence, domains which are described by the location of the center of the transmsmbrane domain, with at least ten transmembrane amino acids on each side of the reported central residue(s).
Proteins and protein fragments of the present invention include proteins with amino acid sequence lengths that are at least 25%(more preferably at least 50%, and most preferably at least 75%) of the length of a disclosed protein and have at least 60% sequence identity (more preferably, at least 75% identity; most preferably at least 90%
or 95%
identity) with that disclosed protein, where sequence identity is determined by comparing the amino acid sequences of the proteins when aligned so as to maximize overlap and identity while minimizing sequence gaps. Also included in the present invention are proteins and protein fragments that contain a segment preferably comprising 8 or more (more preferably 20 or more, most preferably 30 or more) contiguous amino 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
2 0 (Washington University BLAST) version 2.0 software, which builds upon WU-BLAST
version 1.4, which in turn is based on the public domain NCBI-BLAST version 1.4 (Altschul and Gish, 1996, Local alignment statistics, Doolittle ed., Methods in Enzymology 266: 460-480; Altschul et al., 1990, Basic local alignment search tool, Journal of Molecular Biology 215: 403-410; Gish and States, 1993, Identification of protein coding 2 5 regions by database similarity search, Nature Genetics 3: 266-272; Karlin and Altschul, 1993, Applications and statistics for multiple high-scoring segments in molecular sequences, Proc. Natl. Acad. Sci. USA 90: 5873-5877; all of which are incorporated by reference herein). WU-BLAST version 2.0 executable programs for several UNIX
platforms can be downloaded from ftp:/lblast.wustl.edu/blast/executables. The complete 3 0 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 TBL,ASTX -- 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 turned off in all of these programs, if desired. The default penalty (Q) for a gap of length one is Q=9 for proteins and BLASTP, and Q=10 for BLASTN, but may be changed to any integer value including zero, one through eight, nine, ten, eleven, twelve through twenty;
twenty-one through fifty, fifty-one through one hundred, etc. The default per-residue penalty for extending a gap (R) is R=2 for proteins and BLASTP, and R=10 for BLASTN, but may be changed to any integer value including zero, one, two, three, four, five, six, seven, eight, nine, ten, eleven, twelve through twenty, twenty-one through fifty, fifty-one through one hundred, etc. Any combination of values for Q and R can be used in order to align sequences so as to maximize overlap and identity while minimizing sequence gaps.
The default amino acid comparison matrix is BLOSUM62, but other amino acid comparison matrices such as PAM can be utilized.
Species homologues of the disclosed polynucleotides and proteins are also provided by the present invention. As used herein, a "species homologue" is a protein or polynucleotide with a different species of origin from that of a given protein or polynucleotide, but with significant sequence similarity to the given protein or 2 0 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 2 5 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 3 0 species. Most preferably, species homologues are those isolated from certain mammalian species such as, for example, Pan troglodytes, Gorilla gorilla, Pongo pygmaeus, Hylobates concolor, Maraca mulatta, Papio papio, Papio ~amadryas, Cercopithecus aethiops, Cebus capucinus, Aotus trivirgatus, Sanguinus Oedipus, Microcebus murinus, Mus musculus, Rattus norvegicus, Cricetulus griseus, Fells catus, 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 Seu4nez, 1988, Ann. Rev. Genet. 22: 323-351; O'Brien et al., 1993, Nature Genetics 3:103-112; Johansson et al.,1995, Genomics 25: 682-690; Lyons et al.,1997, Nature Genetics 15: 47-56; O'Brien et al.,1997, Trends in Genetics 13(10): 393-399;
Carver and Stubbs, 1997, Genome Research 7:1123-1137; all of which are incorporated by reference herein).
The invention also encompasses allelic variants of the disclosed polynudeotides or proteins; that is, naturally-occurring alternative forms of the isolated polynucleotides which also encode proteins which are identical or have significantly similar sequences to those encoded by the disclosed 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 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.
2 0 The invention also includes polynucleotides with sequences complementary to those of the polynucleotides disclosed herein.
The present invention also includes polynucleotides that hybridize under reduced stringency conditions, more preferably stringent conditions, and most preferably highly stringent conditions, to polynucleotides described herein. Examples of stringency 2 5 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.

WO 99/32614 PCT/US98n7140 StringencyPolynucleotideHybridHybridization TemperatureWash ConditionHybrid Lengthand Temperature ~p)= Buffer' and 8uffert 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 x 50 67C; lxSSC -or- 67C; 0.3xSSC
45C; lxSSC, 50% formamide D DNA:RNA <50 Tp*; lxSSC TD*; lxSSC

E RNA:RNA Z 50 70C; lxSSC -or- 70C; 0.3xSSC
50C;IxSSC,50~oformanude F RNA:RNA <50 TF*; lxSSC TF*; lxSSC

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

I DNA:RNA 2 50 67C;4xSSC-or- 67C; lxSSC
45C; 4xSSC, 50~ formamide 1 DNA:RNA <50 T~*; 4xSSC T~*; ,~~

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

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

O DNA:RNA 250 55C;4xSSC-or- 55C;2xSSC
42C; 6xSSC, 50Yo formamide P DNA:RNA <50 TP*; 6xSSC TP*; 6xSSC

Q RNA:RNA 2 50 60C; 4xSSC -or- (~0C;
45C; 6xSSC, 509 formamide 2 R R1VA:ItNA <50 TR*; 4xSSC TR*: 4xSSC

t: The hybrid length is that anticipated for the hybridized regions) of the hybridizing polynucleotides. When hybridizing a polynucleatide to a target polynucleotide of unknown sequence, the hybrid length is assumed to be that of the hybridizing polynucieotide. When polynucleotides of known sequence are hybridized, the 2 5 hybrid length can be defiamined by aligning the sequences of the polynucleotides and identifying the region or regions of optimal sequence complementarity.
': SSPE (lxSSPE is 0.15M NaCI, lOmM NaHiPO" 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 "Te- T~: 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 (T°,) of the hybrid, where Tm is determined according to the following equations. For hybrids less than 18 base pairs in length, T",(°C) = 2(ii of A + T bases) + 4(# of G +
C bases). For hybrids between 18 and 49 base pairs in l~gth, 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 [Na') is the concentration of sodium ions in the 3 5 hybridization buffer ([Na'] for lxSSC = 0.165 M).

Additional examples of stringency conditions for polynucleotide hybridization are provided in Sambrook, J., E.F. Fritsch, and T. Maniatis, 1989, Molecular Cloning: A
Laboratory Manual, Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY, chapters 9 and 11, and Current Protocols in Molecular Biology,1995, F.M.
Ausubel et al., eds., john Wiley & Sons, Inc., sections 2.10 and 6.3-6.4, incorporated herein by reference.
Preferably, each such hybridizing polynucleotide has a length that is at least 25%(more preferably at least 50%, and most preferably at least 75%) of the length of the polynucleotide of the present invention to which it hybridizes, and has at least 6(?%
sequence identity (more preferably, at least 75% identity; most preferably at least 90% or 95% identity) with the polynucleotide of the present invention to which it hybridizes, where sequence identity is determined by comparing the sequences of the hybridizing polynucleotides when aligned so as to maximize overlap and identity while minimizing sequence gaps.
The isolated polynucleotide of the invention may be operably linked to an expression control sequence such as the pMT2 or pED expression vectors disclosed in Kaufman et al., Nucleic Acids Res. ,~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$~, 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- Mamnnalian 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 i_ri 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 rnay 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 MaxBacO kit), and such methods are weU known in the art, as described in Summers and Smith, Texas Agricultural Exnerimgnt Station Bulletin N~. 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 Sepharose~; 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 (TRH. Kits for expression and purification of such fusion proteins are commercially available from New England BioLabs (Beverly, MA), Pharmacia (Piscataway, NJ) and 3 0 Invitrogen Corporation (Carlsbad, CA), respectively. The protein can also be tagged with an epitope and subsequently purified by using a specific antibody directed to such epitope. One such epitope ("Flag") is commercially available from the Eastman Kodak Company (New Haven, CT).

Finally, one or more reverse-phase high performance liquid chromatography (1ZP-I-iPLC) steps employing hydrophobic,RP-HPLC media, e.g., silica gel having pendant methyl or other aliphatic groups, can be employed to further purify the protein. Some or all of the foregoing purification steps, in various combinations, can also be employed to provide a substantially homogeneous isolated recombinant protein. The protein thus purified is substantially free of other mammalian proteins and is defined in accordance with the present invention as an "isolated protein."
The protein of the invention may also be expressed as a product of transgenic animals, e.g., as a component of the milk of transgenic cows, goats, pigs, or sheep which are characterized by somatic or germ cells containing a nucleotide sequence encoding the protein.
The protein may also be produced by known conventional chemical synthesis.
Methods for constructing the proteins of the present invention by synthetic means are known to those skilled in the art. The synthetically-constructed protein sequences, by virtue of sharing primary, secondary or tertiary structural and/or conformational characteristics with proteins may possess biological properties in common therewith, including protein activity. Thus, they may be employed as biologically active or imrnunological substitutes for natural, purified proteins in screening of therapeutic compounds and in immunological processes for the development of antibodies.
2 0 The proteins provided herein also include proteins characterized by amino acid sequences similar to those of purified proteins but into which modification are naturally provided or deliberately engineered. For example, modifications in the peptide or DNA
sequences can be made by those skilled in the art using known techniques.
Modifications of interest in the protein sequences may include the alteration, substitution, replacement, 2 5 insertion or deletion of a selected amino acid residue in the coding sequence. For example, one or more of the cysteine residues may be deleted or replaced with another amino acid to alter the conformation of the molecule. Techniques for such alteration, substitution, replacement, insertion or deletion are well known to those skilled in the art (see, e.g., U.S. Patent No. 4,518,584). Preferably, such alteration, substitution, replacement, 3 0 insertion or deletion retains the desired activity of the protein.
Other fragments and derivatives of the sequences of proteins which would be expected to retain protein activity in whole or in part and may thus be useful for screening or other immunological methodologies may also be easily made by those skilled in the art given the disclosures herein. Such modifications are believed to be encompassed by the present invention.
USES AND BIOLOGICAL ACTIVITY
The polynucleotides and proteins of the present invention are expected to exhibit one or more of the uses or biological activities (including those associated with assays cited herein) identified below. Uses or activities described for proteins of the present invention may be provided by administration or use of such proteins or by administration or use of polynucleotides encoding such proteins (such as, for example, in gene therapies or vectors suitable for introduction of DNA).
Research 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, Cell 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.
Polynucleotides and proteins of the present invention can also be used as nutritional sources or supplements. . Such uses include without limitation use as a protein 2 5 or amino acid supplement, use as a carbon source, use as a nitrogen source and use as a source of carbohydrate. In such cases the protein or polynucleotide of the invention can be added to the feed of a particular organism or can be administered as a separate solid or liquid preparation, such as in the form of powder, pills, solutions, suspensions or capsules. In the case of microorganisms, the protein or polynucleotide of the invention 3 0 can be added to the medium in or on which the microorganism is cultured.
Cytokine aid Cell Proli_fPra+;r", ~~~erentiation cti~~iri~
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, T10, B9, B9/I1, BaF3, MC9/G, M+ {preB M+), 2E8,.RB5, 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.1~5:1706-1712, 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. Vol 1 pp. 3.12.1-3.12.14, john Wiley and Sons, Toronto. 1994; and Measurement of mouse and human Interferon y, Schreiber, R.D. In Current Protocols in Immunology. J.E.e.a. Coligan eds. Vol 1 pp. 6.8.1-6.8.8, john Wiley and Sons, Toronto.1994.
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 Interleukin 11- Bennett, F., Giannotti, J., Clark, S.C. and Turner, K. J. In Current Protocols pCT/US98/27140 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. 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; Chapter 6, Cytokines and their cellular receptors; Chapter 7, Imrnunologic studies in Humans);
Weinberger et al., Proc. Natl. Acad. Sci. USA 77:6091-6095, 1980; Weinberger et al., Eur. J.
Immure.

11:405-411,1981; Takai et al., J. Immunol.137:3494-3500,1986; Takai et al., J.
l.mmunol.
140:508-512, 1988.
ini~iune Stimulati_r~Q or Sunvressi_ng Activity A protein of the present invention may also exhibit immune stimulating or immune suppressing activity, including without limitation the activities for which assays are described herein. A protein may be useful in the treatment of various immune 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 autoimmune 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. Of course, in this regard, a protein of the present invention may also be useful where a boost to the immune system generally may be desirable, i.e., in the 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, autoinunune pulmonary inflammation, Guillain-Barre syndrome, autoimmune thyroiditis, insulin dependent diabetes mellitis, myasthenia gravis, graft versus-host disease and autoimmune inflammatory eye disease.
Such a protein of the present invention may also to be useful in the treatment of allergic reactions and conditions, such as asthma (particularly allergic asthma) or other respiratory problems. Other conditions, in which immune suppression is desired (including, for example, organ transplantation), may also be treatable using a protein of the present invention.
Using the proteins of the invention it may also be possible to regulate immune responses in a number of ways. Down regulation may be in the form of inhibiting or blocking an immune response already in progress or may involve preventing the induction of an immune response. The functions of activated T cells may be inhibited by suppressing T cell responses or by inducing specific tolerance in T cells, or both.
Immunosuppression of T cell responses is generally an active, non-antigen-specific, process which requires continuous exposure of the T cells to the suppressive agent.
Tolerance, which involves inducing non-responsiveness or anergy in T cells, is distinguishable from immunosuppression in that it is generally antigen-specific and persists after exposure to the tolerizing agent has ceased. Operationally, tolerance can be demonstrated by the lack of a T cell response upon reexposure to specific antigen in the absence of the tolerizing agent.
Down regulating or preventing one or more antigen functions (including without 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 in Immunology. J.E.e.a. Coligan eds.

anergize the T cells, thereby inducing tolerance in a subject. Induction of long-term tolerance by B lymphocyte antigen-blocking reagents may avoid the necessity of repeated administration of these blocking reagents. To achieve sufficient immunosuppression or tolerance in a subject, it may also be necessary to block the function of a combination of B lymphocyte antigens.
The efficacy of particular blocking reagents in preventing organ transplant rejection or GVI ID can be assessed using animal models that are predictive of efficacy in hun;tans. 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 GVI3D (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/lpr/Ipr 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. Far 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 I5 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 (eg., 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 aMivity. The transfected tumor cells are returned to the patient to result in expression of the peptides on the surface of the transfected cell. Alternatively, gene therapy techniques can be used to target a tumor cell for transfection in vivo.
The presence of the peptide of the present invention having the activity of a B
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 transfiected 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-domaizi 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); Hernnann et al., Proc. Natl.
Acad. Sci.
2 0 USA 78:2488-2492,1981; Herrrnann 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., J. 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. Ixnmunol. 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 Thl and CTL responses) include, without limitation, WO 99/32614 PCT/US98n7140 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.
Hematovoiesis R~~ulating Activi 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 4?

precursors and/or erythroid cells; in supporting the growth and proliferation of myeloid cells such as granulocytes and monocytes/macrophages (i.e., traditional CSF
activity) useful, for example, in conjunction with chemotherapy to prevent or treat consequent myelo-suppression; in supporting the growth and proliferation of megakaryocytes and consequently of platelets thereby allowing prevention or treatment of various platelet disorders such as thrombocytopenia, and generally for use in place of or complimentary to platelet transfusions; and/or in supporting the growth and proliferation of hematopoietic stem cells which are capable of maturing to any and all of the above-mentioned hematopoietic cells and therefore find therapeutic utility in various stem cell disorders (such as those usually treated with transplantation, including, without limitation, aplastic anemia and paroxysmal nocturnal hemoglobinuria), as well as in repopulating the stem cell compartment post irradiation/chemotherapy, either in-vivo or ex-vivo (i.e., in conjunction with bone marrow transplantation or with peripheral progenitor cell transplantation (homologous or heterologous)) as normal cells or genetically manipulated for gene therapy.
The activity of a protein of the invention may, among other means, be measured by the following methods:
Suitable assays for proliferation and differentiation of various hematopoietic lines are cited above.
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 al. eds. Vol pp. l-21, Wiley-Liss, Inc.., New York, NY.1994; Long term bone marrow cultures in the presence of stromal cells, Spooncer, E., Dexter, M. and Allen, T. In Culture of Hematopoietic Cells. R.I. Freshney, et al. eds. Vol pp. 163-179, Wiley-Liss, Inc., New York, NY.1994; Long term culture initiating cell assay, Sutherland, H.J. In Culture of Hematopoietic Cells. R.I. Freshney, et al. eds. Vol pp.139-162, Wiley-Liss, lnc., New York, NY.1994.
Tissue Growth ctivitv 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/ligarnent-like tissue inducing protein may have prophylactic use in preventing damage to tendon or ligament tissue, as well as use in the improved fixation of tendon or ligament to bone or other tissues, and in repairing defects to tendon or ligament tissue. De novo tendon/ligament-like tissue formation induced by a composition of the present invention contributes to the repair of congenital, trauma induced, or other tendon or ligament defects of other origin, and is also useful in cosmetic plastic surgery far 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 WO 99132614 PCTIUS98n7140 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, ~nidermal_ Wound HeaLne, 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).
Activi_n~/Inhibin Activity A protein of the present invention may also exhibit activin- or inhibin-related activities. Inhibins are characterized by their ability to inhibit the release of follicle stimulating hormone (FSH), while activins and are characterized by their ability to stimulate the release of follicle stimulating hormone (FSH). Thus, a protein of the present 2 5 invention, alone or in heterodimers with a member of the inhibin a family, may be useful as a contraceptive based on the ability of inhibins to decrease fertility in female mammals and decrease spermatogenesis in male mammals. Administration of sufficient amounts of other inhibins can induce infertility in these mammals. Alternatively, the protein of the invention, as a homodimer or as a heterodimer with other protein subunits of the inhibin-3 0 [i group, may be useful as a fertility inducing therapeutic, based upon the ability of activin molecules in stimulating FSH release from cells of the anterior pituitary.
See, for example, United States Patent 4,798,885. A protein of the invention may also be useful for advancement of the onset of fertility in sexually immature mammals, so as to increase the lifetime reproductive performance of domestic animals such as cows, sheep and pigs.

The activity of a protein of the invention may, among other means, be measured by the following methods:
Assays for activin/inlu'bin 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.
(''hpmntac~tir /C'hpmt~kineti~ ACtIVitV
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, f'broblasts, neutrophils, T-cells, mast cells, eosinophils, epithelial and/or endothelial cells.
Chemotactic and chemokinetic proteins can be used to mobilize or attract a desired cell population to a desired site of action. Chemotactic or chemokinetic proteins provide particular advantages in treatment of wounds and other trauma to tissues, as well as in treatment of localized infections. For example, attraction of lymphocytes, monocytes or neutrophils to tumors or sites of infection may result in improved immune responses against the tumor or infecting agent.
A protein or peptide has chemotactic activity for a particular cell population if it can stimulate, directly or indirectly, the directed orientation or movement of such cell 2 0 population. Preferably, the protein or peptide has the ability to directly stimulate directed movement of cells. Whether a particular protein has chemotactic activity for a population of cells can be readily determined by employing such protein or peptide in any known assay for cell chemotaxis.
The activity of a protein of the invention may, among other means, be measured 2 5 by the following methods:
Assays for chemotactic activity {which will identify proteins that induce or prevent chemotaxis) consist of assays that measure the ability of a protein to induce the migration of cells across a membrane as well as the ability of a protein to induce the adhesion of one cell population to another cell population. Suitable assays for movement and adhesion 3 0 include, without limitation, those described in: Current Protocols in Immunology, Ed by J.E. Coligan, A.M. Kruisbeek, D.H. Margulies, E.M. Shevach, W.Strober, Pub.
Greene Publishing Associates and Wiley Interscience (Chapter 6.12, Measurement of alpha and beta Chemokines 6.12.1-6.12.28; Taub et al. J. Clin. Invest. 95:1370-1376,1995; Lind et al.

APMIS 103:140-146,1995; Muller et al Eur. J. Immunol. 25:1744-1748; Gruber et al. J. of Immunol.152:5860-5867,1994; Johnston et al. J. of Immunol. 153:1762-1768,1994.
Hemostatic and Thromb~;~tic Activ:it~r A protein of the invention may also exhibit hemostatic or thrombolytic activity.
As a result, such a protein is expected to be useful in treatment of various coagulation disorders (including hereditary disorders, such as hemophilias) or to enhance coagulation and other hemostatic events in treating wounds resulting from trauma, surgery or other causes. A protein of the invention may also be useful for dissolving or inhibiting formation of thromboses and for treatment and prevention of conditions resulting therefrom (such as, for example, infarction of cardiac and central nervous system vessels (e.g., stroke).
The activity of a protein of the invention may, among other means, be measured by the following methods:
Assay for hemostatic and thrombolytic activity include, without limitation, those described in: Linet et al., J. Clin. Pharmacol. 26:131-140,1986; Burdick et al., Thrombosis Res. 45:413-419,1987; Humphrey et al., Fibrinolysis 5:71-79 (1991); Schaub, Prostaglandins 35:467-474,1988.
2 0 R~tor/L~and Activitv 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-In_~a_mmatorX Activity Proteins of the present invention may also exhibit anti-inflammatory activity.
The anti-inflammatory activity may be achieved by providing a stimulus to cells involved in the inflammatory response, by inhibiting or promoting cell-cell interactions (such as, for example, cell adhesion), by inhibiting or promoting chemotaxis of cells involved in the inflammatory process, inhibiting or promoting cell extravasation, or by stimulating or suppressing production of other factors which more directly inhibit or promote an inflammatory response. Proteins exhibiting such activities can be used to treat inflammatory conditions including chronic or acute conditions), including without limitation inflammation associated with infection (such as septic shock, sepsis or systemic 2 0 inflammatory response syndrome (SIRS)), ischemia-reperfusion injury, endotoxin lethality, arthritis, complement-mediated hyperacute rejection, nephritis, cytokine or chemokine-induced lung injury, inflammatory bowel disease, Crohn's disease or resulting from over production of cytokines such as TNF or IL-1. Proteins of the invention may also be useful to treat anaphylaxis and hypersensitivity to an antigenic substance or material.
radheri_n /Tumor Ixivasion Suppressor 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 WO 99!32614 PCTIUS98I27140 extracellular repeats (cadherin domains), but structural differences are found in other parts of the molecule. The cadherin domains bind calcium to form their tertiary structure and thus calcium is required to mediate their adhesion. Only a few amino acids in the first cadherin domain provide the basis for homophilic adhesion; modification of this recognition site can change the specificity of. a cadherin so that instead of recognizing only itself, the mutant molecule can now also bind to a different cadherin. In addition, some cadherins engage in heterophilic adhesion with other cadherins.
E-cadherin, one member of the cadherin superfamily, is expressed. in epithelial cell types. Pathologically, if E-cadherin expression is lost in a tumor, the malignant cells become invasive and the cancer metastasizes. Transfection of cancer cell lines with polynucleotides expressing E-cadherin has reversed cancer-associated changes by returning altered cell shapes to normal, restoring cells' adhesiveness to each other and to their substrate, decreasing the cell growth rate, and drastically reducing anchorage-independent cell growth. Thus, reintroducing E-cadherin expression reverts carcinomas to a less advanced stage. It is likely that other cadherins have the same invasion suppressor role in carcinomas derived from other tissue types. Therefore, proteins of the present invention with cadherin activity, and polynucleotides of the present invention encoding such proteins, can be used to treat cancer. Introducing such proteins or polynucleotides into cancer cells can reduce or eliminate the cancerous changes observed 2 0 in these cells by providing normal cadherin expression.
Cancer cells have also been shown to express cadherins of a different tissue type than their origin, thus allowing these cells to invade and metastasize in a different tissue in the body. Proteins of the present invention with cadherin activity, and polynucleotides of the present invention encoding such proteins, can be substituted in these cells for the 2 5 inappropriately expressed cadherins, restoring normal cell adhesive properties and reducing or eliminating the tendency of the cells to metastasize.
Additionally, proteins of the present invention with cadherin activity, and polynucleotides of the present invention encoding such proteins, can used to generate antibodies recognizing and binding to cadherins. Such antibodies can be used to block 3 0 the adhesion of inappropriately expressed tumor-cell cadherins, preventing the cells from forming a tumor elsewhere. Such an anti-cadherin antibody can also be used as a marker for the grade, pathological type, and prognosis of a cancer, i.e. the more progressed the cancer, the less cadherin expression there will be, and this decrease in cadherin expression can be detected by the use of a cadherin-binding antibody.

Fragments of proteins of the present invention with cadherin activity, preferably a polypeptide comprising a decapeptide of the cadherin recognition site, and poly-nucleotides of the present invention encoding such protein fragments, can also be used to block cadherin function by binding to cadherins and preventing them from binding in ways that produce undesirable effects. Additionally, fragments of proteins of the present invention with cadherin activity, preferably truncated soluble cadherin fragments which have been found to be stable in the circulation of cancer patients, and polynucleotides encoding such protein fragments, can be used to disturb proper cell-cell adhesion.
Assays for cadherin adhesive and invasive suppressor activity include, without limitation, those described in: Hortsch et al. J Biol Chem 270 (32): 18809-18817, 1995;
Miyaki et al. Oncogene 11: 2547 2552,1995; Ozawa et al. Cell 63:1033-1038,1990.
Tumor Inhibition Activii~
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 characteristic, 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 DOSIrf~C' 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 earner) diluents, fillers, salts, buffers, stabilizers, solubilizers, and other materials well known in the art. The term "pharmaceutically acceptable" means a non-toxic material that does not interfere with the 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, TIVF, IL-1, IL-2, IL-3, IL-4, IL-5, IL-6, II,-7, ):L~g, I~,_9, n;10, IL-11, ILrl2, ILrl3,1Ir14, 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.

WO 99132614 PC"T/US98/27140 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 carriers, 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, lympholcines or other hematopoietic factors. When co-administered with one or more cytolcines, lymphokines or other hematopoietic factors, protein of the present invention may be administered either simultaneously with the cytolcine(s), lympholcine(s), other hematopoietic factor(s), thrombolytic or anti-thrombotic factors, or sequentially. If administered sequentially, the attending physician will decide on the appropriate sequence of administering protein of the present invention in combination with cytolcine(s), lympholcine{s), other hematopoietic factor(s), thrombolytic or anti-thrombotic factors.
Administration of protein of the present invention used in the pharmaceutical composition or to practice the method of the present invention can be carned 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, end preferably from about 25 to 90% protein of the present invention.
When administered in liquid form, a liquid carrier 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 contains from about 0.5 to 90% by weight of protein of the present invention, and preferably from about 1 to 50% protein of the present invention.

When a therapeutically effeMive 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 pg 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 hemocyanin (KLH). Methods for synthesizing such peptides are known in the art, for example, as in R.P. Merrifield, J. Amer.Chem.Soc. $ ~" 2149-2154 (1963); J.L. Krstenansky, et al., FEBS Lett.
~1_, 10 (198. 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. Qther 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.
2 0 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 hydroxyallcylcelluloses), 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 derived 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 WO 99/32614 PCTIUS98l27140 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.

WO 99132614 PCTNS98l27140 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.
Wong, Gordon G.
Clark, Hilary Fechtel, Kim Genetics Institute, Inc.
<120> SECRETED PROTEINS AND POLYNUCLEOTIDES ENCODING THEM
<130> 6057A
<140>
<141>
<160> 17 <170> Patentin Ver. 2.0 <210> 1 <211> 4292 <212> DNA
<213> Homo sapiens <400> 1 ctcaagttaa accaacaagc cgatagaaaa aggtagttat casgagattt taaaacttca 60 accctttttc tcttatagtt agtgaagaga gtagaatatc tccagttttg gctgacatct 120 ctacaacctg aacaattggc ttsaacttca cttgggattc ccggttgctt gttttagcat 180 ggcaaaattt ggcgttcaca gaatccttct tctggctatt tctctgacaa agtgtctgga 240 gagtacaaaa ctgctggcag accttaaaaa atgtggtgac ttggaatgtg aagctttaat 300 aaacagagtc tcagccatga gagattatag aggacctgac tgccgatacc tgaacttcac 360 taagggagaa gagatatctg tttatgttaa acttgcagga gaaagggaag atttgtgggc 420 aggaagtaaa ggaaaggagt ttggatattt tcccagagat gcagtccaga ttgaagaggt 480 gttcatatct gaggaaattc agatgtcaac gaaagaatct gactttcttt gtcttcttgg 540 agtaagttac acatttgaca atgasgatag tgaattaaac ggtgattatg gtgaaaatat 600 atatccttat gaagaagata aagatgaaaa atctagtata tatgaaagtg attttcagat 660 agaacctgga ttttatgcaa cttatgaaag tactttgttt gaagaccaag ttccagcatt 720 agaggctcct gaagatatcg gaagtaccag tgaatcaaaa gactgggaag aagtagttgt 780 tgaaagtatg gaacaggatc gtattccaga agtgcatgtc ccaccatctt cagctgtgtc 840 tggagtcaaa gaatggtttg gattgggagg agancaagct gaagagaagg cttttgaatc 900 agttattgaa cctgtacaag aaagctcatt tcggagtaga aaaatagcag tggaagatga 960 gaatgaccta gaggaattaa ataatggtga gcctcaaaca gaacatcagc aagaatctga 1020 atcagaaatt gattcagtgc caaagacaca gtctgaacta gcatctgagt cagagcacat 1080 tcccaaacct caatccactg gttggtttgg tggaggattt acaagttatt taggttttgg 1140 agatgaggat acagggcttg aattaatagc tgaagaaagc aatccaccac tacaagattt 1200 tcccaatccc atatcatctg ataaagaagc cacagttcca tgtacagaaa tattaacaga 1260 aaaaaaagac acaatcacta atgatagctt gagtctcaag ccaagttggt ttgattttgg 1320 ttttgctata ctaggctttg catatgccaa ggaagataaa attatgttag atgacaggaa 1380 aaatgaagaa gatggtgggg cagatgaaca tgaacatcct ctaacaagtg aattagaccc 1440 tgaaaaagaa caagaaatag aaacgataaa aattatagaa acagaagatc aaatagacaa 1500 gaaaccagtc tcagaaaaaa cagacgaatc tgatactata ccatatttga aaaagttctt 1560 gtataatttt gacaaccctt ggaacttcca gaacattcca aaggaaacag aattgccatt 1620 SUBSTITUTE SKEET (RULE 28) WO 99/32614 PCT/ItS98/27140 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gaggggtttt cctccttacc ttcccccaag acctggattt ttccccccac ccccacattc 4200 tgaaggtaga agtgagttcc cctcaggttt gattccacct tcaaatgagc ctgctactga 4260 acatccagaa ccacagcaag aaacctgaca at 4292 <210> 2 <211> 1369 <212> pRT
<213> Homo sapiens <400> 2 Met Ala Lys Phe Gly Val His Arg Ile Leu Leu Leu Ala Ile Ser Leu Thr Lys Cys Leu Glu Ser Thr Lys Leu Leu Ala Asp Leu Lys Lys Cys Gly Asp Leu Glu Cys Glu Ala Leu Ile Asn Arg Val Ser Ala Met Arg i Asp Tyr Arg Gly Pro Asp Cys Arg Tyr Leu Asn Phe Thr Lys Gly Glu Glu Ile Ser Val Tyr Val Lys Leu Ala Gly Glu Arg Glu Asp Leu Trp Ala Gly Ser Lys Gly Lys Glu Phe Gly Tyr Phe Pro Arg Asp Ala Val Gln Ile Glu Glu Val Phe Ile Ser Glu Glu Ile Gln Met Ser Thr Lys Glu Ser Asp Phe Leu Cys Leu Leu Gly Val Ser Tyr Thr Phe Asp Asn Glu Asp Ser Glu Leu Asn Gly Asp Tyr Gly Glu Asn Ile Tyr pro Tyr Glu Glu Asp Lys Asp Glu Lys Ser Ser Ile Tyr Glu Ser Asp Phe Gln Ile Glu Pro Gly Phe Tyr Ala Thr Tyr Glu Ser Thr Leu Phe Glu Asp Gln Val Pro Ala Leu Glu Ala Pro Glu Asp Ile Gly Ser Thr Ser Glu Ser Lys Asp Trp Glu Glu Val Val Val Glu Ser Met Glu Gln Asp Arg Ile Pro Glu Val His Val Pro Pro Ser Ser Ala Val Ser Gly Val Lys Glu Trp Phe Gly Leu Gly Gly Glu Gln Ala Glu Glu Lys Ala Phe Glu Ser Val Ile Glu Pro Val Gln Glu Ser Ser Phe Arg Ser Arg Lys Ile Ala Val Glu Asp Glu Asn Asp Leu Glu Glu Leu Asn Asn Gly Glu Pro Gln Thr Glu His Gln Gln Glu Ser Glu Ser Glu Ile Asp Ser Val Pro Lys Thr Gln Ser Glu Leu Ala Ser Glu Ser Glu His Ile Pro Lys Pro Gln Ser Thr Gly Trp Phe Gly Gly Gly Phe Thr Ser Tyr Leu Gly Phe Gly Asp Glu Asp Thr Gly Leu Glu Leu Ile Ala Glu Glu Ser Asn pro Pro Leu Gln Asp Phe Pro Asn Pro Ile Ser Ser Asp Lys Glu Ala Thr Val Pro Cys Thr Glu Ile Leu Thr Glu Lys Lys Asp Thr Ile Thr Asn Asp Ser Leu Ser Leu Lys Pro Ser Trp Phe Asp phe Gly Phe Ala Ile Leu Gly Phe Ala Tyr Ala Lys Glu Asp Lys Ile Met Leu Asp Asp Arg Lys Asn Glu Glu Asp Gly Gly Ala Asp Glu His Glu His Pro Leu Thr Ser Glu Leu Asp pro Glu Lys Glu Gln Glu Ile Glu Thr Ile Lys Ile Ile Glu Thr Glu Asp Gln Ile Asp Lys Lys pro Val Ser Glu Lys Thr Asp Glu Ser Asp Thr Ile Pro Tyr Leu Lys Lys Phe Leu Tyr Asn Phe Asp Asn Pro Trp Asn Phe Gln Asn Ile Pro Lys Glu Thr Glu Leu Pro Phe Pro Lys Gln Ile Leu Asp Gln Asn Asn Val Ile Glu Asn Glu Glu Thr Gly Glu Phe Ser Ile Asp Asn Tyr Pro Thr Asp Asn Thr Lys Val Met Ile Phe Lys Ser Ser Tyr Ser Leu Ser Asp Met Val Ser Asn Ile Glu Leu Pro Thr Arg Ile His Glu Glu Val Tyr Phe Glu Pro Ser Ser Ser Lys Asp Ser Asp Glu Asn Ser Lys Pro Ser Val Asp Thr Glu Gly Pro Ala Leu Val Glu Ile Asp Arg Ser Val Glu Asn Thr Leu Leu Asn Ser Gln Met Val Ser Thr Asp Asn Ser Leu Ser Ser Gln Asn Tyr Ile Ser Gln Lys Glu Asp Ala Ser Glu Phe Gln Ile Leu Lys Tyr Leu Phe Gln Ile Asp Val Tyr Asp phe Met Asn Ser Ala Phe Ser Pro Ile Val Ile Leu Thr Glu Arg Val Val Ala Ala Leu Pro Glu Gly Met Arg Pro Asp Ser Asn Leu Tyr Gly Phe pro Trp Glu Leu Val Ile Cys Ala Ala Val Val Gly Phe Phe Ala Val Leu Phe Phe Leu Trp Arg Ser Phe Arg Ser Val Arg Ser Arg Leu Tyr Val Gly Arg Glu Lys Lys Leu Ala Leu WO 99/32614 PCT/US98n7140 Met Leu Ser Gly Leu Ile Glu Glu Lys Ser Lys Leu Leu Glu Lys Phe Ser Leu Val Gln Lys Glu Tyr Glu Gly Tyr Glu Val Glu Ser Ser Leu Lys Asp Ala Ser Phe Glu Lys Glu Ala Thr Glu Ala Gln Ser Leu Glu Ala Thr Cys Glu Lys Leu Asn Arg Ser Asn Ser Glu Leu Glu Asp Glu Ile Leu Cys Leu Glu Lys Glu Leu Lys Glu Glu Lys Ser Lys His Ser Glu Gln Asp Glu Leu Met Ala Asp Ile Ser Lys Arg Ile Gln Ser Leu Glu Asp Glu~Ser Lys Ser Leu Lys Ser Gln Val Ala Glu Ala Lys Met Thr Phe Lys Ile PhP Gln Met Asn Glu Glu Arg Leu Lys Ile Ala Ile Lys Asp Ala Leu Asn Glu Asn Ser Gln Leu Gln Glu Ser Gln Lys Gln Leu Leu Gln Glu Ala Glu Val Trp Lys Glu Gln Val Ser Glu Leu Asn Lys Gln Lys Val Thr Phe Glu Asp Ser Lys Val His Ala Glu Gln Val Leu Asn Asp Lys Glu Ser His Ile Lys Thr Leu Thr Glu Arg Leu Leu Lys Met Lys Asp Trp Ala Ala Met Leu Gly Glu Asp Ile Thr Asp Asp Asp Asn Leu Glu Leu Glu Met Asn Ser Glu Ser Glu Asn Gly Ala Tyr Leu Asp Asn Pro Pro Lys Gly Ala Leu Lys Lys Leu Ile His Ala Ala Lys Leu Asn Ala Ser Leu Lys Thr Leu Glu Gly Glu Arg Asn Gln Ile Tyr Ile Gln Leu Ser Glu Val Asp Lys Thr Lys Glu Glu Leu Thr Glu His Ile Lys Asn Leu Gln Thr Gln Gln Ala Ser Leu Gln Ser Glu Asn 965 970 g75 Thr His Phe Glu Asn Glu Asn Gln Lys Leu Gln Gln Lys Leu Lys Val Met Thr Glu Leu Tyr Gln Glu Asn Glu Met Lys Leu His Arg Lys Leu WO 99l3Z614 PGT/US98/27140 Thr Val Glu Glu Asn Tyr Arg Leu Glu Lys Glu Glu Lys Leu Ser Lys Val Asp Glu Lys Ile Ser His Ala Thr Glu Glu Leu Glu Thr Tyr Arg Lys Arg Ala Lys Asp Leu Glu Glu Glu Leu Glu Arg Thr Ile His Ser Tyr Gln Gly Gln Ile Ile Ser His Glu Lys Lys Ala His Asp Asn Trp Leu Ala Ala Arg Asn Ala Glu Arg Asn Leu Asn Asp Leu Arg Lys Glu Asn Ala His Asn Arg Gln Lys Leu Thr Glu Thr Glu Leu Lys Phe Glu Leu Leu Glu Lys Asp Pro Tyr Ala Leu Asp Val Pro Asn Thr Ala Phe Gly Arg Gly Ser Arg Gly Pro Gly Asn Pro Leu Asp His Gln Ile Thr Asn Glu Arg Gly Glu Ser Ser Cys Asp Arg Leu Thr Asp Pro His Arg Ala Pro Ser Asp Thr Gly Ser Leu Ser Pro Pro Trp Asp Gln Asp Arg Arg Met Met Phe Pro Pro Pro Gly Gln Ser Tyr Pro Asp Ser Ala Leu Pro Pro Gln Arg Gln Asp Arg Phe Cys Ser Asn Ser Gly Arg Leu Ser Gly Pro Ala Glu Leu Arg Ser Phe Asn Met Pro Ser Leu Asp Lys Met Asp Gly Ser Met Pro Ser Glu Met Glu Ser Ser Arg Asn Asp Thr Lys Asp Asp Leu Gly Asn Leu Asn Val Pro Asp Ser Ser Leu Pro Ala Glu Asn Glu Ala Thr Gly Pro Gly Phe Val Pro Pro Pro Leu Ala Pro Ile Arg Gly Pro Leu Phe Pro Val Asp Ala Arg Gly Pro Phe Leu Arg Arg Gly Pro Pro Phe Pro Pro Pro Pro Pro Gly Ala Met Phe Gly Ala Ser Arg Asp Tyr Phe Pro Pro Arg Asp Phe Pro Gly Pro Pro Pro Ala Pro Phe Ala Met Arg Asn Val Tyr Pro Pro Arg Gly Phe Pro Pro Tyr Leu Pro Pro Arg Pro Gly Phe Phe Pro Pro Pro Pro His Ser Glu Gly Arg Ser Glu Phe Pro Ser Gly Leu Ile Pro Pro Ser Asn Glu Pro Ala Thr Glu His Pro Glu Pro Gln Gln Glu Thr <210> 3 <211> 412 <212> DNA
<213> Homo sapiens <400> 3 ttactttatg ccagatccag tacatgccat tgttatcctc tgtttacagg tggggaagct 60 gaggcaggaa gccctttagt cacttgccga aggccacgct gttacccatg ggaccggttt 120 tgggcggccg aagagcactc atggggccgg attcacgccc cgggcccgtt ccctcctgct 180 ctctggtgct cctcacgcca ttggccccac tgcctctcac tgcccgtgag tccctgtgcc 240 cgtgtcctcc ttcttgaacc cctcagccct cagttaaccc tcagaaagct ggctcggaga 300 agtccttgtg tggtatctgg gaggcagagt ttgccgtgag ccgagattgt gccactgcac 360 gcactccagc ctgggcgaca gagcgagacc ccatctcaaa aaaaaaaaaa as 412 <210> 4 <211> 49 <212> PRT
<213> Homo sapiens <400> 4 Met Gly Pro Val Leu Gly Gly Arg Arg Ala Leu Met Gly Pro Asp Ser Arg Pro Gly Pro Val Pro Ser Cys Ser Leu Val Leu Leu Thr Pro Leu Ala Pro Leu Pro Leu Thr Ala Arg Glu Ser Leu Cys Pro Cys Pro Pro Ser <210> 5 <211> 1317 <212> DNA
<213> Homo sapiens <400> 5 gtctgtgaga gtcaattcag gggaaagata caagattgat ttgtaaaacc cttgaaatgt 60 agatttcttg tagatgtatc cttcacgttg taaatatgtt ttgtagagtg aagccatggg 120 aagccatgtg taacagagct tagacatcca aaactaatca atgctgaggt ggctaaatac 180 ctagcctttt acatgtaaac ctgtctgcaa aattagcttt tttaaaaaaa aaaaaaaaaa 240 aaaattgggg gggttaattt atcattcaga aatcttgcat tttcaaaaat tcagtgcaag 300 cgccaggcga tttgtgtcta aggatacgat tttgaaccat atgggcagtg tacaaaatat 360 gaaacaactg tttccacact tgcacctgat caagagcagt gcttctccat ttgttttgca 420 gagaaatgtt tttcatttcc cgtgtgtttc catttccttc tgaaattctg attttatcca 480 tttttttaag gctcctcttt atctcctttc ttaaggcact gttgctatgg cacttttcta 540 taaccttttc attcctgtgt acagtagctt aaaattgcag tgattgagca taacctactt 600 WO 99132614 PCTIUS98lZ7140 gtttgtataa attattgaaa tccatttgca ccctgtaaga atggacttaa aagtactgct 660 ggacaggcat gtgtgctcaa agtacattga ttgctcaaat ataaggaaat ggcccaatga 720 acgtggttgt gggaggggaa agaggaaaca gagctagtca gatgtgaatt gtatctgttg 780 taataaacat gttaaaacaa acaaaaattg ttatttttct tttccttcgg tcagtgcaca 840 ttagcatttg aactacctgg ggattcttta tcagaactgt tcttgttgaa tatttatact 900 taattgaaat aattccttaa gggaggtttt gtttaaaacg tattaacagg aaattgtgta 960 tgagatattt aatgaaataa gaaattcaac aagaatgatt aagtcacttc ccaagtggtt 1020 gtcatttgtt aaaccctggt ttacctgtct tgctattatg acatttcatt tggaaggatg 1080 tttgtgttgt agctaactgt tcaagtctgg tgctgactgc tgttcttagc catcacasaa 1140 cgctaaattt gtgtaattgg agcttcctgc tgttatctgg aaatagcagg aaagcgcagc 1200 tttgtatatt gtttcctaaa gtatattaaa ataaaaaaag aaactattgc tactaaaaaa 1260 aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaa 1317 <210> 6 <211> 4B
<212> PRT
<213> Homo sapiens <400> 6 Met Phe Phe Ile Ser Arg Val Phe Pro Phe Pro Ser Glu Ile Leu Ile Leu Ser Ile Phe Leu i~rg Leu Leu Phe Ile Ser Phe Leu Lys Ala Leu Leu Leu Trp His Phe Ser Ile Thr Phe Ser Phe Leu Cys Thr Val Ala <210> 7 <211> 1442 <212> DNA
<213> Homo sapiens <400> 7 tgcggttgtt ttccttctct ccgtgcsacg ctggcaagtc tcaaagtcgc cacagaaaca 60 tgcccctgat tcagtgcctc tgcttagctg taacatgtta atcagaacta cctggcatct 120 tcctgaacaa gactttcaat aggggccagt atgcttcgct tcatccagaa gttttctcaa 180 gcatcttcaa agatactgaa gtactctttc ccagtgggac taagaaccag cagaacagat 240 atactttctc tcaagatgtc tctccagcaa aacttttccc catgtccaag gccttggctt 300 tcctcatcat ttccagcgta tatgagcaag acacagtgct atcatacatc cccctgcagc 360 tttaaaaagc agcagaagca agcacttcta gccagaccct caagcaccat cacttaccta 420 actgacagcc caaagccagc attatgtgta actctggcag gactaatccc cttcgttgct 480 ccaccactgg tcatgctgat gacaaasact tatattccca tattagcttt tactcagatg 540 gcttatggag ccagtttcct atctttcttg ggtgggatca gatggggttt tgctctacca 600 gaaggtagtc cagccaaacc agactacctt aatttagcta gcagtgcagc tcctcttttc 660 ttttcatggt ttgccttcct tatttctgaa agacttagtg aagccatagt cacagtaata 720 atgggtatgg gagtagcatt ccaccttgaa ctttttctct taccacatta tcccaactgg 780 tttaaagccc tgaggatagt agtcacttta ttggccactt tttcatttat aatcacttta 840 gtagttaaaa gtagttttcc agaaaaagga cataagagac ctggtcaagt atasaaaata 900 taaaagtctg ggaagtgagg agcacctctg cccagctgct gccccgtctg ggaagtgagg 960 agcgcctctg cctggccgcc tgaccatctg ggaagtgtga caagcgcctc tgcccggccg 1020 ctgtgcaacc ttccacgtgt gaagtgacag ccttgtgtgt gatcttttct gtcttcccca 1080 agtttgcatt ttcgacatta aagtttactt tttagttaaa agttttaaaa atatatatat 1140 ataaatacac tgtagataac atttgtatgc cagctacacc tttttctact tctgtttggc 1200 tttttttccc cacaccaatg gtaatttatc ttcacagatt gttcttcatt tctagaaatt 1260 gttacttcat ggtaattact tgagcaaaag cttgaaaatc cctgacaagt acttttcatc 1320 g tcatagtata ttagttttca ctcagtcatt ttatgaataa tatagttatc cacttaaaca 1380 tttcaatatt ttaaccatct tgaaaattaa agattaaaaa tccccttaaa saaaaaaaaa 1440 as <210> 8 <211> 247 <212> PRT
<213> Homo sapiens <400> 8 Met Leu Arg Phe Ile Gln Lys Phe Ser Gln Ala Ser Ser Lys Ile Leu Lys Tyr Ser Phe Pro Val Gly Leu Arg Thr Ser Arg Thr Asp Ile Leu Ser Leu Lys Met Ser Leu GIn Gln Asn Phe Ser Pro Cys Pro Arg Pro Trp Leu Ser Ser Ser Phe Pro Ala Tyr Met Ser Lys Thr Gln Cys Tyr His Thr Ser Pro Cys Ser Phe Lys Lys Gla Gln Lys Gln Ala Leu Leu Ala Arg pro Ser Ser Thr Ile Thr Tyr Leu Thr Asp Ser Pro Lys Pro Ala Leu Cys Val Thr Leu Ala Gly Leu Ile Pro Phe Val Ala Pro Pro loo las llo Leu Val Met Leu Met Thr Lys Thr Tyr Ile Pro Ile Leu Ala Phe Thr Gln Met Ala Tyr Gly Ala Ser Phe Leu Ser Phe Leu Gly Gly Ile Arg Trp Gly Phe Ala Leu Pro Glu Gly Ser Pro Ala Lys Pro Asp Tyr Leu Asn Leu Ala Ser Ser Ala Ala Pro Leu Phe Phe Ser Trp Phe Ala Phe Leu Ile Ser Glu Arg Leu Ser Glu Ala Ile Val Thr Val Ile Met Gly Met Gly Val Ala Phe His Leu Glu Leu Phe Leu Leu Pro His Tyr Pro Asn Trp Phe Lys Ala Leu Arg Ile Val Val Thr Leu Leu Ala Thr Phe Ser Phe Ile Ile Thr Leu VaI Val Lys Ser Ser Phe Pro Glu Lys Gly His Lys Arg pro Gly Gln Val <210> 9 <211> 2696 <212> DNA
<213> Homo sapiens <400> 9 taggcctctt tggccgtgga gctgtccctc ctaggtggga atgttcactc ttttgttgtc 60 aggttgtatg ggggggcagg gacagtgctg ctgggaagga tgccagctct gggattgggc 120 cagtcctgtg ggcaagactt gcaagaggct ggatcaactt ggtgtggtat ctctgatggc 180 ttagagtaat ggcaatgagg gtctctgttg tgatgtcact gagtactttc tggggtgctt 240 ctgggcaccc cttatgatgt cacaggaagc agttcctcag aggttacttc ctgtgaacat 300 aagggagcag gtacttcctg tgatgtctca atgagttctt ccttgaaggt cactttggtg 360 atatcatggg aaggtgtact tccggtgatg cctagaggtc acgtcctgtg atgtcattag 420 gctgaagcat gtacttcctg tattggacag tgaccagtct ctgacctgcc ttctccctcc 480 acacccttct ttggtgggtg ttggcctggg ggtcttccta ggaagagaat aaggcacggg 540 acttggatcc aatctggagg actctaacgg aaaaaaacca atattgtcag ggtcatctcc 600 attcaaacac gtcaagtctg cgaactcgcc ctggagggag gggtgggaga tggacctagt 660 gcaaactact gttaaagacc tcccttccca cccctgcctt ttgtgtgcat gcctgtgtct 720 gcgtggcttt gtttcattga atcgggtgag ccaagtgtgt ggtggctctg tcaggccagt 780 atggccaggt gtagagttca gtgagccata gtagggtccc ttgggccaga atgcttcgtg 840 tggtctgata ggttagattg gtttggtggc ttccaccaag ctggcatact tcggtgatgt 900 ctgatggatc agaatgtttt ggtgtgctca ccaggggctc tggagaacta gaatgttctg 960 atggagtctg acaagccagg cggccttgag agttggttta ggagtggctc cctgagtgct 1020 gtggctgtgg tcagctctaa ggacctgttg gcagactgag attt~agggc ctgacaacca 1080 tgtagactag gatagctgag acctccctct acccccaccc atctccctct cttccttgga 1140 gaccacctcc actttctcca acccaaagca gggcgcccag tgccctggtt ccatcatcag 1200 catctctggg ggaggggcgc cccatgccca ccctctcccc catttgtccg cctcctaggt 1260 cttccaascc cttttcttct ctatgacttt gggggaaatc cagcctcctt gtctctctcc 1320 taaaaaagga gggaagaaaa gccacagaga caattcctgc ccctaaagcc taggagatcc 1380 ctctcccttg ctagagagcc acccccaaat caaaatgtga aaatccctag aaagcaatag 1440 ccttcgaggt accttgcact gaatttccca ccccagccct tccacccgat gggaggctgt 1500 aacttgggca ctggggtgac tttttccatg cccttgtcat ctccagggtg ggaggcaggc 1560 cccacttccc ctcccctatc ccccacttcc cattgttgtt gccccacccc taatctccag 1620 actgaaccca gatggagatc tgagtgccaa aacaattctt gatgtaactt tgtacatatc 1680 ttctactacc gttgggggct cttggggtta gaggtggggg cggctctgtg ggccattgct 1740 cccctccacc tctcaaaaga ccttacagta tttcacagta tctctacccg cacgcgagta 1800 ttacagtatc tagctggaat atccccctac agccccccag gaccctatga ggaagggaag 1860 gagccaggga gagtgaagta aggtctggga ctggggaggt gggatctgat gaactcattt 1920 gcatatcatt cgcatcctcc gcttggcagc cgctttctac aaactcattc actggagtct 1980 gggtcccaat cagccgggtc caggactcct ctcacacaga cacatctccg gaggctgggc 2040 ctcctgaaaa gtgtttgctt ggggtgtctg tgtaacaacc cctccctatt catatttctt 2100 ggggaccccc tacccagcca gccagggtga tctgaaaggt atactttgct agctcagtga 2160 gctagttcac tcaccatgtt ggtgagcaga gagccacacc tttccccatt ttaccttggg 2220 aaactcactc caccatcttt gccatctctt gaaagtccct tctgcaatct gacctcaatc 2280 ttttgtgctg cagtttgtcc agaggggaca cagatgtggg gtcagggatg aggattattg 2340 aaaaacccat catctctttt ttttttcccc gtctccctat tagccaatcc gatctcagag 2400 tctctgagtg gcctccttgc accttctctt cagcacccag taggtgctta ataagtgttt 2460 gctgcattga attatctccc tattccttct catttgccct ctagcttccc ataccttctc 2520 caagtgtctt cctccctttc tttgtctggc tccctatgac tttctatttt tttttcctcc 2580 gtgtggttcc cattgttttc tgtcctgtct ctatcttagt ctttgtctgt cttcctcctt 2640 tcctcaaatg tctcaactct ctctccccaa tttccccatt taaaaaaaaa aaaaaa 2696 <210> 10 <211> 73 <212> PRT
<213> Homo sapiens <400> 10 Met Asn Ser Phe Ala Tyr His Ser His Pro Pro Leu Gly Ser Arg Phe 1~

WO 99/32614 PCTNS98n7140 Leu Gln Thr His Ser Leu Glu Ser Gly Ser Gln Ser Ala Gly Ser Arg Thr Pro Leu Thr Gln Thr His Leu Arg Arg Leu Gly Leu Leu Lys Ser Val Cys Leu Gly Cys Leu Cys Asn Asn Pro Ser Leu Phe Ile Phe Leu Gly Asp Pro Leu Pro Ser Gln Pro Gly 65 7p <210> 11 <211> 2791 <212> DNA
<Z13> Homo sapiens <400> 11 tttttttaga gtggtaacat ttattaggaa ggagagactt cagttagact cccatgtctc 60 acacagaaga aacagatggt aactgccatt tctttctttc tttctttctt tttttttttg 120 agatggagtt tcgctcttat tgcccaggct ggagtgcaat ggtgcaattt cggctcacct 180 caacctccgc ctcccggttt caagagattc tcctgcctca gcctcctaag tagctgggat 240 tacaggcata cgccaccatg cctggctaat tctgtatttt tagtagacac ggggtttctc 300 catgttggtc aggctggtct tgaactcccg acctcaggtg atccgcccac ctcggcttcc 360 caaagtgctg ggattacaag cgtgagccac tgcgcccagc cagtaactgc catttctaaa 420 gaggaaagag agcaggcaga gggtcctgac tcccagggga caggtagttc agctggacaa 480 tgagggagta tgagattagg gtggataagg acactgctca ccaccctcca ctgaagttca 540 gtggctaaaa tactgctaca ccagccaatc agtggaaggc tatcttgctc tccaggggac 600 acttgggatg tggttggtgg caggaagaaa gaatgtcatg ctatctcttg ctgctcctcc 660 gggtttcttt gccgttacta acagggttgc tggaagggcc aggcgggcct gcaggtgtgt 720 ggttgggctg gcttcgagtg attcgggtgc tgggtgggtg ggagggagta tgagggggat 780 gtgggccacc accgggacct ggtggggcac tcagtccatt cccattctgg ttctcagagg 840 ctggagaggc agcagagggt ccagtggtag gctgggaagc tgaagggttg ctgtcgttct 900 gcaccgcctt gttgggtgcc tgctgggtgc tgtactctgg gtttggctca ctgaagttag 960 gaacctcaga ataasccata cagaattctc caatcttctg cagatcaccc ccacggccag 1020 tatatagtgt cagacaacct ttgaaaactg aagcgtaccc tttggtgagc cggataatgt 1080 ccccaggctg gatcagattg ccaacatcgt cccagacaga gatattgatg ctgcctgttt 1140 tgtccgccac tttgcaggtc cgaacctcat gcccgtcctt tgtcttggtc actcggcctg 1200 tctccagcac aatgaagata aggttcagat tcttgagccc aggcttgata tccttcacaa 1260 aggtctccgt cgtcatgctg cctgagcctc ggcaccccac tggatgggga atagaggatg 1320 aaggctcaat gcttggatcc tttgccccca tgctccactt cccaaggcct acctacccca 1380 ttaggatggg gtcaggcagc ctcaacccct ctaacccttc taaaagatta aaaaagaaca 1440 tccccggagg cctccaactt caggatcaga atttgggggt ctctggacag gctgctttgg 1500 ggctggaagg ccccctgccg ggatgctctt ttagtctcaa gccgcgaagt ggcggagccg 1560 acgtagacag gggtagggaa cccggtgcac agccagggag tagaatctta ctggccagat 1620 cttccggaac cctcatcccg acaagccggg actcggtcca tcccctcccc taccggcagc 1680 ccaccaccca cccaagccag ccggcaggac tgtgccgtgg ctggaagtta ctgtgaggcg 1740 gcggctaaga aggcggttct ggtggcggcg gtggaggctg aggcggcggc cgaggcggcg 1800 acggaggaaa cagaagatgg cagatttttt gaaaggactg cctgtctaca acaaaagcaa 1860 ttttagtcga tttcacgcgg actccgtgtg caaagcctcg aaccgacggc cctcagtcta 1920 cctgcctacc cgcgagtacc cgtctgaaca gatcatcgtg acagaaaaga caaacatcct 1980 cctgcgctac ctgcatcagc aatgggacaa aaagaacgct gccaagaaga gagaccagga 2040 gcaagtggag ctcgaaggcg agagctccgc acctccccgc aaggtggcgc ggaccgacag 2100 cccagacatg cacgaggaca cttaagactc tcaactccac aggcgcctcc tgccaggtct 2160 gctcctcggt cgcccacccg cctgcccgcc atgtgtaagc accccgcccg cccgcctccc 2220 tgccggccca tccacaccct gcgtccacac cacttccaac ctcataggag ccgatgtatt 2280 tattttcctt gagtttttat ttatgctgta acctgtatca agcgttggtt aaaggggaca 2340 tcagacccag tagtgtgatg ttggtagatg ctttttaaaa aaaacaacat tgtccccccg 2400 acccccgcct tccatcgggc cagttccccg attcctgccc ccagttctcc agagaaccag 2460 agtgtgtctg tgagagtctc tagcgggggc tttactgtgg ccgggcgaca ggggcgggcc 2520 cggggtggcc tgacctacca ggacagccga gtggccttct cccccccaac accgatccag 2580 gccattgaga ctcggtcttg tcccacgttc gcccggaact ttcccatgcc cagacctcac 2640 tcagcgtgca cgcacgttgg ggagaagtcg gcccttggga tctttctctt gagtcatttt 2700 atttttatca tggactagtg cgtgctccgt gtccacccca ataaaagggt ctttcctaaa 2760 aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa a 2791 <210> 12 <211> 219 <212> PRT
<213> Homo sapiens <400> 12 Met Ser Pro Gly Trp Ile Arg Leu Pro Thr Ser Ser Gln Thr Glu Ile Leu Met Leu Pro Val Leu Ser Ala Thr Leu Gln Val Arg Thr Ser Cys Pro Ser Phe Val Leu Val Thr Arg Pro Val Ser Ser Thr Met Lys Ile Arg Phe Arg Phe Leu Ser Pro GIy Leu Ile Ser Phe Thr Lys Val Ser Val Val Met Leu Pro Glu Pro Arg His Pro Thr Gly Trp Gly Ile Glu Asp Glu Gly Ser Met Leu Gly Ser Phe Ala Pro Met Leu His Phe Pro Arg Pro Thr Tyr Pro Ile Arg Met Gly Ser Gly Ser Leu Asn Pro Ser Asn Pro Ser Lys Arg Leu Lys Lys Asn Ile Pro Gly Gly Leu Gln Leu Gln Asp Gln Asn Leu Gly Val Ser Gly Gln Ala Ala Leu Gly Leu Glu Gly Pro Leu Pro Gly Cys Ser Phe Ser Leu Lys Pro Arg Ser Gly Gly Ala Asp Val Asp Arg Gly Arg Glu Pro Gly Ala Gln Pro Gly Ser Arg Ile Leu Leu Ala Arg Ser Ser Gly Thr Leu Ile Pro Thr Ser Arg Asp Ser Val His Pro Leu Pro Tyr Arg Gln Pro Thr Thr His Pro Ser Gln Pro Ala Gly Leu Cys Arg Gly Trp Lys Leu Leu <210> 13 <211> 29 <212> DNA
<213> Artificial Sequence <220>
<223> oligonucleotide <220>
<221> misc_feature <222> (2) <223> biotinylated phosphoaramidite residue «400> 13 cnaattgttc aggttgtaga gatgtcagc 29 <210> 14 <211> 29 <212> DNA
<213> Artificial Sequence <220>
<223> oligonucleotide <220>
<221> misc_feature <222> (2) <223> biotinylated phosphoaramidite residue «400> 14 tnagaaggaa atggaaacac acgggaaat <210> 15 <211> 29 <212> DNA
<213> Artificial Sequence <220>
<223> oligonucleotide <220>
<221> misc_feature <222> (2) <223> biotinylated phosphoaramidite residue «400> 15 tnagcatgac cagtggtgga gcaacgaag 29 <210> 16 <211> 20 <212> DNA
<213> Artificial Sequence <220>
<223> oligonucleotide <400> 16 ggtatgggaa gctagagggc ~ 20 <210> 17 <211> 18 <212> DNA
<213> Artificial Sequence <220>
<223> oligonucleotide <400> 17 gtctgggacg atgttggc 18

Claims (29)

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 179 to nucleotide 4285;
(c) a polynucleotide comprising the nucleotide sequence of the full-length protein coding sequence of clone am728_60 deposited under accession number ATCC 98621;
(d) a polynucleotide encoding the full-length protein encoded by the cDNA insert of clone am728_60 deposited under accession number ATCC 98621;
(e) a polynucleotide encoding a protein comprising the amino acid sequence of SEQ ID NO:2;
(f) 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;
(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).

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 am728_60 deposited under accession number ATCC 98621.

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 am728_60 deposited under accession number ATCC 98621;
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; and (ab) the nucleotide sequence of the cDNA insert of clone am728_60 deposited under accession number ATCC 98621;
(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; and (bb) the nucleotide sequence of the cDNA insert of clone am728_60 deposited under accession number ATCC 98621;
(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; and (w) a nucleotide sequence corresponding to the cDNA sequence of SEQ ID
NO:1 from nucleotide 179 to nucleotide 4285, and extending contiguously from a nucleotide sequence corresponding to the 5' end of said sequence of SEQ ID
NO:1 from nucleotide 179 to nucleotide 4285, to a nucleotide sequence corresponding to the 3' end of said sequence of SEQ ID NO:1 from nucleotide 179 to nucleotide 4285.

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 108 to nucleotide 254;
(c) a polynucleotide comprising the nucleotide sequence of SEQ ID
NO:3 from nucleotide 225 to nucleotide 254;

(d) a polynucleotide comprising the nucleotide sequence of the full-length protein coding sequence of clone bf377_1 deposited under accession number ATCC 98621;
(e) a polynucleotide encoding the full-length protein encoded by the cDNA insert of clone bf377_1 deposited under accession number ATCC 98621;
(f) a polynucleotide comprising the nucleotide sequence of the mature protein coding sequence of clone bf377_1 deposited under accession number ATCC 98621;
(g) a polynucleotide encoding the mature protein encoded by the cDNA insert of clone bf377_1 deposited under accession number ATCC 98621;
(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 bf377_1 deposited under accession number ATCC 98621;
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 bf377_1 deposited under accession number ATCC 98621;
(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: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 bf377_1 deposited under accession number ATCC 98621;
(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 108 to nucleotide 254, and extending contiguously from a nucleotide sequence corresponding to the 5' end of said sequence of SEQ ID
NO:3 from nucleotide 108 to nucleotide 254, to a nucleotide sequence corresponding to the 3' end of said sequence of SEQ ID NO:3 from nucleotide 108 to nucleotide 254;
and (x) a nucleotide sequence corresponding to the cDNA sequence of SEQ ID
NO:3 from nucleotide 225 to nucleotide 254, and extending contiguously from a nucleotide sequence corresponding to the 5' end of said sequence of SEQ ID
NO:3 from nucleotide 225 to nucleotide 254, to a nucleotide sequence corresponding to the 3' end of said sequence of SEQ ID NO:3 from nucleotide 225 to nucleotide 254.

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 426 to nucleotide 569;
(c) a polynucleotide comprising the nucleotide sequence of SEQ ID
NO:5 from nucleotide 546 to nucleotide 569;
(d) a polynucleotide comprising the nucleotide sequence of the full-length protein coding sequence of clone cw354_1 deposited under accession number ATCC 98621;
(e) a polynucleotide encoding the full-length protein encoded by the cDNA insert of clone cw354_1 deposited under accession number ATCC 98621;
(f) a polynucleotide comprising the nucleotide sequence of the mature protein coding sequence of clone cw354_1 deposited under accession number ATCC 98621;
(g) a polynucleotide encoding the mature protein encoded by the cDNA insert of clone cw354_1 deposited under accession number ATCC 98621;
(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 cw354_1 deposited under accession number ATCC 98621;
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 cw354_1 deposited under accession number ATCC 98621;
(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 cw354_1 deposited under accession number ATCC 98621;
(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 426 to nucleotide 569, and extending contiguously from a nucleotide sequence corresponding to the 5' end of said sequence of SEQ ID
NO:5 from nucleotide 426 to nucleotide 569, to a nucleotide sequence corresponding to the 3' end of said sequence of SEQ ID NO:5 from nucleotide 426 to nucleotide 569;
and (x) a nucleotide sequence corresponding to the cDNA sequence of SEQ ID
NO:5 from nucleotide 546 to nucleotide 569, and extending contiguously from a nucleotide sequence corresponding to the 5' end of said sequence of SEQ ID
NO:5 from nucleotide 546 to nucleotide 569, to a nucleotide sequence corresponding to the 3' end of said sequence of SEQ ID NO:5 from nucleotide 546 to nucleotide 569.

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 151 to nucleotide 891;
(c) a polynucleotide comprising the nucleotide sequence of SEQ ID
NO:7 from nucleotide 595 to nucleotide 891;
(d) a polynucleotide comprising the nucleotide sequence of the full-length protein coding sequence of clone nm134 4 deposited under accession number ATCC 98621;
(e) a polynucleotide encoding the full-length protein encoded by the cDNA insert of clone nm134_4 deposited under accession number ATCC 98621;
(f) a polynucleotide comprising the nucleotide sequence of the mature protein coding sequence of clone nm134_4 deposited under accession number ATCC 98621;
(g) a polynucleotide encoding the mature protein encoded by the cDNA insert of clone nm134_4 deposited under accession number ATCC 98621;
(h) a polynucleotide encoding a protein comprising the amino acid sequence of SEQ ID NO:8;

(i) 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;
(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).

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) the amino acid sequence of SEQ ID NO:8 from amino acid 104 to amino acid 163;
(c) fragments of the amino acid sequence of SEQ ID NO:8, each fragment comprising eight consecutive amino acids of SEQ ID NO:8; and (d) the amino acid sequence encoded by the cDNA insert of clone nm134_4 deposited under accession number ATCC 98621;
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 nm134_4 deposited under accession number ATCC 98621;
(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 nm134_4 deposited under accession number ATCC 98621;
(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;
(w) a nucleotide sequence corresponding to the cDNA sequence of SEQ ID
NO:7 from nucleotide 151 to nucleotide 891, and extending contiguously from a nucleotide sequence corresponding to the 5' end of said sequence of SEQ ID
NO:7 from nucleotide 151 to nucleotide 891, to a nucleotide sequence corresponding to the 3' end of said sequence of SEQ ID NO:7 from nucleotide 151 to nucleotide 891;
and (x) a nucleotide sequence corresponding to the cDNA sequence of SEQ ID
NO:7 from nucleotide 595 to nucleotide 891, and extending contiguously from a nucleotide sequence corresponding to the 5' end of said sequence of SEQ ID
NO:7 from nucleotide 595 to nucleotide 891, to a nucleotide sequence corresponding to the 3' end of said sequence of SEQ ID NO:7 from nucleotide 595 to nucleotide 891.

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 1909 to nucleotide 2127;

(c) a polynucleotide comprising the nucleotide sequence of SEQ ID
NO:9 from nucleotide 2074 to nucleotide 2127;
(d) a polynucleotide comprising the nucleotide sequence of the full-length protein coding sequence of clone yb11_1 deposited under accession number ATCC 98621;
(e) a polynucleotide encoding the full-length protein encoded by the cDNA insert of clone yb11_1 deposited under accession number ATCC 98621;
(f) a polynucleotide comprising the nucleotide sequence of the mature protein coding sequence of clone yb11_1 deposited under accession number ATCC
98621;
(g) a polynucleotide encoding the mature protein encoded by the cDNA insert of clone yb11_1 deposited under accession number ATCC 98621;
(h) a polynucleotide encoding a protein comprising the amino and sequence of SEQ ID NO:10;
(i) 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;
(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).

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 yb11_1 deposited under accession number ATCC 98621;
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 done yb11_1 deposited under accession number ATCC 98621;
(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 yb11_1 deposited under accession number ATCC 98621;
(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;
(w) a nucleotide sequence corresponding to the cDNA sequence of SEQ ID
NO:9 from nucleotide 1909 to nucleotide 2127, and extending contiguously from a nucleotide sequence corresponding to the 5' end of said sequence of SEQ ID
NO:9 from nucleotide 1909 to nucleotide 2127, to a nucleotide sequence corresponding to the 3' end of said sequence of SEQ ID NO:9 from nucleotide to nucleotide 2127; and (x) a nucleotide sequence corresponding to the cDNA sequence of SEQ ID
NO:9 from nucleotide 2074 to nucleotide 2127, and extending contiguously from a nucleotide sequence corresponding to the 5' end of said sequence of SEQ ID
NO:9 from nucleotide 2074 to nucleotide 2127, to a nucleotide sequence corresponding to the 3' end of said sequence of SEQ ID NO:9 from nucleotide to nucleotide 2127.

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 1077 to nucleotide 1733;
(c) a polynucleotide comprising the nucleotide sequence of SEQ ID
NO:11 from nucleotide 1158 to nucleotide 1733;
(d) a polynucleotide comprising the nucleotide sequence of the full-length protein coding sequence of clone yc2_1 deposited under accession number ATCC 98621;
(e) a polynucleotide encoding the full-length protein encoded by the cDNA insert of clone yc2_1 deposited under accession number ATCC 98621;
(f) a polynucleotide comprising the nucleotide sequence of the mature protein coding sequence of clone yc2_1 deposited under accession number ATCC
98621;
(g) a polynucleotide encoding the mature protein encoded by the cDNA insert of clone yc2_1 deposited under accession number ATCC 98621;
(h) a polynucleotide encoding a protein comprising the amino acid sequence of SEQ ID NO:12;
(i) 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;
(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).

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) fragments of the amino acid sequence of SEQ ID NO:12, each fragment comprising eight consecutive amino acids of SEQ ID NO:12; and (c) the amino acid sequence encoded by the cDNA insert of clone yc2_1 deposited under accession number ATCC 98621;
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 yc2_1 deposited under accession number ATCC 98621;
(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 yc2_1 deposited under accession number ATCC 98621;
(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;
(w) a nucleotide sequence corresponding to the cDNA sequence of SEQ ID
NO:11 from nucleotide 1077 to nucleotide 1733, and extending contiguously from a nucleotide sequence corresponding to the 5' end of said sequence of SEQ ID
NO:11 from nucleotide 1077 to nucleotide 1733, to a nucleotide sequence corresponding to the 3' end of said sequence of SEQ ID NO:11 from nucleotide 1077 to nucleotide 1733; and (x) a nucleotide sequence corresponding to the cDNA sequence of SEQ ID
NO:11 from nucleotide 1158 to nucleotide 1733, and extending contiguously from a nucleotide sequence corresponding to the 5' end of said sequence of SEQ ID
NO:11 from nucleotide 1158 to nucleotide 1733, to a nucleotide sequence corresponding to the 3' end of said sequence of SEQ ID NO:11 from nucleotide 1158 to nucleotide 1733.