WO1997039123A2 - Secreted proteins - Google Patents

Secreted proteins Download PDF

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Publication number
WO1997039123A2
WO1997039123A2 PCT/US1997/006139 US9706139W WO9739123A2 WO 1997039123 A2 WO1997039123 A2 WO 1997039123A2 US 9706139 W US9706139 W US 9706139W WO 9739123 A2 WO9739123 A2 WO 9739123A2
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WO
WIPO (PCT)
Prior art keywords
polynucleotide
seq
protein
amino acid
acid sequence
Prior art date
Application number
PCT/US1997/006139
Other languages
French (fr)
Other versions
WO1997039123A3 (en
Inventor
Kenneth Jacobs
John M. Mccoy
Lisa A. Racie
Edward R. Lavallie
David Merberg
Vikki Spaulding
Original Assignee
Genetics Institute, Inc.
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Application filed by Genetics Institute, Inc. filed Critical Genetics Institute, Inc.
Priority to EP97922304A priority Critical patent/EP0912731A2/en
Priority to AU28016/97A priority patent/AU2801697A/en
Priority to JP53727097A priority patent/JP2001509004A/en
Publication of WO1997039123A2 publication Critical patent/WO1997039123A2/en
Publication of WO1997039123A3 publication Critical patent/WO1997039123A3/en

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    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
    • C07K14/46—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from vertebrates
    • C07K14/47—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from vertebrates from mammals
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P17/00—Drugs for dermatological disorders
    • A61P17/06—Antipsoriatics
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P25/00—Drugs for disorders of the nervous system
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P29/00—Non-central analgesic, antipyretic or antiinflammatory agents, e.g. antirheumatic agents; Non-steroidal antiinflammatory drugs [NSAID]
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00—Antineoplastic agents
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P5/00—Drugs for disorders of the endocrine system

Definitions

  • the present invention provides novel proteins, along with therapeutic, diagnostic and research utilities for these proteins.
  • the present invention provides a composition comprising an isolated protein encoded by a polynucleotide selected from the group consisting of:
  • such polynucleotide comprises the nucleotide sequence of SEQ ID NO: l from nucleotide 70 to nucleotide 505; the nucleotide sequence of the full length protein coding sequence of clone AP162 deposited under accession number ATCC 98026 ; or the nucleotide sequence of the mature protein coding sequence of clone API 62 deposited under accession number ATCC 98026 .
  • the polynucleotide encodes the full length or mature protein encoded by the cDNA insert of clone AP162 deposited under accession number ATCC 98026 .
  • such polynucleotide encodes a protein comprising the amino acid sequence of SEQ ID NO:2 from amino acid 42 to amino acid 61.
  • the present invention provides a composition comprising a protein, wherein said protein comprises an amino acid sequence selected from the group consisting of:
  • the present invention provides a composition comprising an isolated protein encoded by a polynucleotide selected from the group consisting of:
  • such polynucleotide comprises the nucleotide sequence of SEQ ID NO:4 from nucleotide 230 to nucleotide 791; the nucleotide sequence of SEQ ID NO:4 from nucleotide 31 1 to nucleotide 791 ; the nucleotide sequence of the full length protein coding sequence of clone AM931 deposited under accession number ATCC 98026 ; or the nucleotide sequence of the mature protein coding sequence of clone AM931 deposited under accession number ATCC 98026 .
  • the polynucleotide encodes the full length or mature protein encoded by the cDNA insert of clone AM931 deposited under accession number ATCC 98026 . In yet other preferred embodiments, such polynucleotide encodes a protein comprising the amino acid sequence of SEQ ID NO:5 from amino acid 32 to amino acid 51.
  • the present invention provides a composition comprising a protein, wherein said protein comprises an amino acid sequence selected from the group consisting of: (a) the amino acid sequence of SEQ ID NO:5;
  • protein comprises the amino acid sequence of SEQ ID NO:5 or the amino acid sequence of SEQ ID NO:5 from amino acid 32 to amino acid 51.
  • the present invention provides a composition comprising an isolated protein encoded by a polynucleotide selected from the group consisting of:
  • such polynucleotide comprises the nucleotide sequence of SEQ ID NO 6 from nucleotide 14 to nucleotide 491 , the nucleotide sequence of SEQ ID NO 6 from nucleotide 83 to nucleotide 491 , the nucleotide sequence of the full length protein coding sequence of clone AM610 deposited under accession number ATCC 98026 , or the nucleotide sequence of the mature protein coding sequence of clone AM610 deposited under accession number ATCC 98026
  • the polynucleotide encodes the full length or mature protein encoded by the cDNA insert of clone AM610 deposited under accession number ATCC 98026
  • such polynucleotide encodes a protein comprising the amino acid sequence of SEQ ID NO 7 from ammo acid 31 to ammo acid 50
  • the present invention provides a composition comprising a protein, wherein said protein comprises an ammo acid sequence selected from the group consisting of
  • amino acid sequence encoded by the cDNA insert of clone AM610 deposited under accession number ATCC 98026 the protein being substantially free from other mammalian proteins
  • amino acid sequence encoded by the cDNA insert of clone AM610 deposited under accession number ATCC 98026 the protein being substantially free from other mammalian proteins
  • such protein comprises the ammo acid sequence of SEQ ID NO 7 or the ammo acid sequence of SEQ ID NO 7 from ammo acid 31 to ammo acid 50
  • the present invention provides a composition comprising an isolated protein encoded by a polynucleotide selected from the group consisting of (a) a polynucleotide comprising the nucleotide sequence of SEQ ID NO.9,
  • such polynucleotide comp ⁇ ses the nucleotide sequence of SEQ ID NO 9 from nucleotide 1 to nucleotide 483, the nucleotide sequence of the full length protein coding sequence of clone AM340 deposited under accession number ATCC 98026 , or the nucleotide sequence of the mature protein coding sequence of clone AM340 deposited under accession number ATCC 98026
  • the polynucleotide encodes the full length or mature protein encoded by the cDNA insert of clone AM340 deposited under accession number ATCC 98026
  • such polynucleotide encodes a protein comprising the ammo acid sequence of SEQ ID NO 9 from nucleotide 1 to nucleotide 483, the nucleotide sequence of the full length protein coding sequence of clone AM340 deposited under accession number ATCC 98026 , or the nucleotide sequence of the mature protein coding sequence of clone AM340 deposited
  • the present invention provides a composition comprising a protein, wherein said protein comp ⁇ ses an amino acid sequence selected from the group consisting of (a) the amino acid sequence of SEQ ID NO: 10,
  • the present invention provides a composition comprising an isolated protein encoded by a polynucleotide selected from the group consisting of
  • such polynucleotide comprises the nucleotide sequence of SEQ ID NO 1 1 from nucleotide 15 to nucleotide 462, the nucleotide sequence of SEQ ID NO.11 from nucleotide 87 to nucleotide 462, the nucleotide sequence of the full length protein coding sequence of clone AM282 deposited under accession number ATCC 98026 , or the nucleotide sequence of the mature protein coding sequence of clone AM282 deposited under accession number ATCC 98026
  • the polynucleotide encodes the full length or mature protein encoded by the cDNA insert of clone AM282 deposited under accession number ATCC 98026
  • such polynucleotide encodes a protein comprising the ammo acid sequence of SEQ ID NO 12 from ammo acid 28 to amino acid 47
  • the present invention provides a composition comprising a protein, wherem said protein comprises an amino acid sequence selected from the group consisting of
  • the present invention provides a composition comp ⁇ sing an isolated protein encoded by a polynucleotide selected from the group consisting of
  • such polynucleotide comprises the nucleotide sequence of SEQ ID NO 14 from nucleotide 185 to nucleotide 519, the nucleotide sequence of SEQ ID NO 14 from nucleotide 260 to nucleotide 519, the nucleotide sequence of the full length protein coding sequence of clone AK647 deposited under accession number ATCC 98026 , or the nucleotide sequence of the mature protein coding sequence of clone AK647 deposited under accession number ATCC 98026
  • the polynucleotide encodes the full length or mature protein encoded by the cDNA insert of clone AK647 deposited under accession number ATCC 98026
  • such polynucleotide encodes a protein comprising the ammo acid sequence of SEQ ID NO- 15 from amino acid 27 to ammo acid 46
  • the present invention provides a composition comprising a protein, wherein said protein comprises an ammo acid sequence selected from the group consisting of
  • the protein comprises the ammo acid sequence of SEQ ID NO 15 or the amino acid sequence of SEQ ID NO 15 from amino acid 27 to ammo acid 46
  • the present invention provides a composition comp ⁇ sing an isolated protem encoded by a polynucleotide selected from the group consisting of
  • such polynucleotide comprises the nucleotide sequence of SEQ ID NO 17 from nucleotide 257 to nucleotide 536, the nucleotide sequence of SEQ ID NO 17 from nucleotide 329 to nucleotide 536; the nucleotide sequence of the full length protein coding sequence of clone AK583 deposited under accession number ATCC 98026 ; or the nucleotide sequence of the mature protein coding sequence of clone AK583 deposited under accession number ATCC 98026
  • the polynucleotide encodes the full length or mature protein encoded by the cDNA insert of clone AK583 deposited under accession number ATCC 98026 In yet other preferred embodiments.
  • Such polynucleotide encodes a protein comp ⁇ sing the amino acid sequence of SEQ ID NO 18 from ammo acid 14 to amino acid 33
  • the present invention provides a composition comprising a protein, wherein said protein comprises an ammo acid sequence selected from the group consisting of
  • the present invention provides a composition comprising an isolated protein encoded by a polynucleotide selected from the group consisting of
  • such polynucleotide comprises the nucleotide sequence of SEQ ID NO 20 from nucleotide 179 to nucleotide 476, the nucleotide sequence of the full length protein coding sequence of clone AK533 deposited under accession number ATCC 98026 , or the nucleotide sequence of the mature protein coding sequence of clone AK533 deposited under accession number ATCC 98026
  • the polynucleotide encodes the full length or mature protein encoded by the cDNA insert of clone AK533 deposited under accession number ATCC 98026
  • such polynucleotide encodes a protein comp ⁇ sing the amino acid sequence of SEQ ID NO 21 from amino acid 35 to amino acid 57
  • the present invention provides a composition comprising a protein, wherein said protein comprises an ammo acid sequence selected from the group consisting of
  • amino acid sequence encoded by the cDNA insert of clone AK533 deposited under accession number ATCC 98026 the protein being substantially free from other mammalian proteins
  • the present invention provides a composition comprising an isolated protein encoded by a polynucleotide selected from the group consisting of (a) a polynucleotide comprising the nucleotide sequence of SEQ ID NO:23;
  • polynucleotide which encodes a species homologue of the protein of (h) or (i) above.
  • polynucleotide comprises the nucleotide sequence of SEQ ID NO: 1
  • the polynucleotide encodes the full length or mature protein encoded by the cDNA insert of clone AK296 deposited under accession number ATCC 98026 .
  • such polynucleotide encodes a protein comprising the amino acid sequence of SEQ ID NO:24 from amino acid 81 to amino acid 90.
  • the present invention provides a composition comprising a protein, wherein said protein comprises an amino acid sequence selected from the group consisting of:
  • protein comprises the amino acid sequence of SEQ ID NO:24 or the amino acid sequence of SEQ ID NO:24 from amino acid 81 to amino acid 90.
  • the present invention provides a composition comprising an isolated protein encoded by a polynucleotide selected from the group consisting of: (a) a polynucleotide comprising the nucleotide sequence of SEQ ID NO: (a) a polynucleotide comprising the nucleotide sequence of SEQ ID NO: (a) a polynucleotide comprising the nucleotide sequence of SEQ ID NO:
  • such polynucleotide comprises the nucleotide sequence of SEQ ID NO.26 from nucleotide 58 to nucleotide 655. the nucleotide sequence of the full length protein coding sequence of clone H617 deposited under accession number ATCC 98026 ; or the nucleotide sequence of the mature protein coding sequence of clone H617 deposited uVider accession number ATCC 98026
  • the polynucleotide encodes the full length or mature protein encoded by the cDNA insert of clone H617 deposited under accession number ATCC 98026
  • such polynucleotide encodes a protein comprising the amino acid sequence of SEQ ID NO 27 from amino acid 65 to amino acid 84
  • the present invention provides a composition comp ⁇ sing a protein, wherein said protein comp ⁇ ses an amino acid sequence selected from the group consisting of (a) the amino acid sequence of SEQ ID NO.27,
  • amino acid sequence encoded by the cDNA insert of clone H617 deposited under accession number ATCC 98026 the protein being substantially free from other mammalian proteins
  • the present invention provides a composition comprising an isolated protein encoded by a polynucleotide selected from the group consisting of
  • such polynucleotide comp ⁇ ses the nucleotide sequence of SEQ ID NO 29 from nucleotide 14 to nucleotide 391 , the nucleotide sequence of the full length protein coding sequence of clone BB9 deposited under accession number ATCC 98026 , or the nucleotide sequence of the mature protein coding sequence of clone BB9 deposited under accession number ATCC 98026
  • the polynucleotide encodes the full length or mature protein encoded by the cDNA insert of clone BB9 deposited under accession number ATCC 98026
  • such polynucleotide encodes a protein comprising the amino acid sequence of SEQ ID NO 30 from amino acid 75 to ammo acid 94
  • the present invention provides a composition comprising a protein, wherein said protein comprises an amino acid sequence selected from the group consisting of
  • the present invention provides a composition comprising an isolated protein encoded by a polynucleotide selected from the group consistmg of
  • such polynucleotide comprises the nucleotide sequence of SEQ ID NO 32 from nucleotide 61 to nucleotide 514, the nucleotide sequence of SEQ ID NO 32 from nucleotide 1 15 to nucleotide 514, the nucleotide sequence of the full length protein coding sequence of clone AW191 deposited under accession number ATCC 98026 , or the nucleotide sequence of the mature protein coding sequence of clone AW 191 deposited under accession number ATCC 98026
  • the polynucleotide encodes the full length or mature protein encoded by the cDNA insert of clone AW191 deposited under accession number ATCC 98026
  • such polynucleotide encodes a protein comprising the amino acid sequence of SEQ ID NO:33 from amino acid 24 to amino acid 43.
  • the present invention provides a composition comprising a protein, wherein said protein comprises an amino acid sequence selected from the group consisting of:
  • AW191 deposited under accession number ATCC 98026; the protein being substantially free from other mammalian proteins.
  • the protein comprises the amino acid sequence of SEQ ID NO:33 or the amino acid sequence of SEQ
  • the present invention provides a composition comprising an isolated protein encoded by a polynucleotide selected from the group consisting of:
  • such polynucleotide comp ⁇ ses the nucleotide sequence of SEQ ID NO 35 from nucleotide 180 to nucleotide 525, the nucleotide sequence of SEQ ID NO 35 from nucleotide 339 to nucleotide 525, the nucleotide sequence of the full length protein coding sequence of clone AT211 deposited under accession number ATCC 98026 , or the nucleotide sequence of the mature protein coding sequence of clone AT211 deposited under accession number ATCC 98026
  • the polynucleotide encodes the full length or mature protein encoded by the cDNA insert of clone AT211 deposited under accession number ATCC 98026
  • such polynucleotide encodes a protein comprising the amino acid sequence of SEQ ID NO 36 from amino acid 1 to amino acid 20
  • the present invention provides a composition comp ⁇ sing a protein, wherein said protein comprises an ammo acid sequence selected from the group consisting of (a) the amino acid sequence of SEQ ID NO 36,
  • the protein being substantially free from other mammalian proteins
  • such protein comprises the ammo acid sequence of SEQ ID NO 36 or the ammo acid sequence of SEQ ID NO 36 from amino acid 1 to amino acid 20
  • the present invention provides a composition comprising an isolated protein encoded by a polynucleotide selected from the group consisting of
  • such polynucleotide comprises the nucleotide sequence of SEQ ID NO:38 from nucleotide 225 to nucleotide 677; the nucleotide sequence of SEQ ID NO:38 from nucleotide 390 to nucleotide 677; the nucleotide sequence of the full length protein coding sequence of clone AT205 deposited under accession number ATCC 98026 ; or the nucleotide sequence of the mature protein coding sequence of clone AT205 deposited under accession number ATCC 98026 .
  • the polynucleotide encodes the full length or mature protein encoded by the cDNA insert of clone AT205 deposited under accession number ATCC 98026 . In yet other preferred embodiments, such polynucleotide encodes a protein comprising the amino acid sequence of SEQ ID NO:39 from amino acid 6 to amino acid 25.
  • the present invention provides a composition comprising a protein, wherein said protein comprises an amino acid sequence selected from the group consisting of:
  • the present invention provides a composition comprising an isolated protein encoded by a polynucleotide selected from the group consisting of
  • polynucleotide which encodes a species homologue of the protein of (h) or (l) above
  • such polynucleotide comprises the nucleotide sequence of SEQ ID NO:40 from nucleotide 128 to nucleotide 508; the nucleotide sequence of SEQ ID NO:40 from nucleotide 200 to nucleotide 508; the nucleotide sequence of the full length protein coding sequence of clone AS34 deposited under accession number ATCC 98026 ; or the nucleotide sequence of the mature protein coding sequence of clone AS34 deposited under accession number ATCC 98026 .
  • such polynucleotide encodes a protein comprising the amino acid sequence of SEQ ID NO:41 from amino acid 27 to amino acid 46.
  • the present invention provides a composition comprising a protein, wherein said protein comprises an amino acid sequence selected from the group consisting of:
  • protein comprises the amino acid sequence of SEQ ID NO:41 or the amino acid sequence of SEQ ID NO:41 from amino acid 27 to amino acid 46.
  • the present invention provides a composition comprising an isolated protein encoded by a polynucleotide selected from the group consisting of: (a) a polynucleotide comprising the nucleotide sequence of SEQ ID NO: (a) a polynucleotide comprising the nucleotide sequence of SEQ ID NO: (a) a polynucleotide comprising the nucleotide sequence of SEQ ID NO:
  • such polynucleotide comprises the nucleotide sequence of SEQ ID NO:43 from nucleotide 23 to nucleotide 676; the nucleotide sequence of the full length protein coding sequence of clone AS32 deposited under accession number ATCC 98026 ; or the nucleotide sequence of the mature protein coding sequence of clone AS32 deposited under accession number ATCC 98026 .
  • the polynucleotide encodes the full length or mature protein encoded by the cDNA insert of clone AS32 deposited under accession number ATCC 98026 .
  • such polynucleotide encodes a protein comprising the amino acid sequence of SEQ ID NO:44 from amino acid 78 to amino acid 97.
  • the present invention provides a composition comprising a protein, wherein said protein comprises an amino acid sequence selected from the group consisting of:
  • the present invention provides a composition comprising an isolated protein encoded by a polynucleotide selected from the group consisting of
  • such polynucleotide comprises the nucleotide sequence of SEQ ID NO 46 from nucleotide 132 to nucleotide 479, the nucleotide sequence of SEQ ID NO 46 from nucleotide 201 to nucleotide 479, the nucleotide sequence of the full length protein coding sequence of clone AR260 deposited under accession number ATCC 98026 , or the nucleotide sequence of the mature protein coding sequence of clone AR260 deposited under accession number ATCC 98026
  • the polynucleotide encodes the full length or mature protein encoded by the cDNA insert of clone AR260 deposited under accession number ATCC 98026
  • such polynucleotide encodes a protein comprising the amino acid sequence of SEQ ID NO 47 from ammo acid 40 to amino acid 59
  • the present invention provides a composition comp ⁇ sing a protein, wherem said protein comprises an amino acid sequence selected from the group consisting of
  • protem comprises the amino acid sequence of SEQ ID NO 47 or the amino acid sequence of SEQ
  • the present invention provides a composition comprising an isolated protein encoded by a polynucleotide selected from the group consisting of
  • polynucleotide which encodes a species homologue of the protem of (g) or (h) above
  • polynucleotide comprises the nucleotide sequence of SEQ ID NO: 1
  • polynucleotide encodes the full length or mature protein encoded by the cDNA insert of clone K640 deposited under accession number ATCC 98026
  • polynucleotide encodes a protein comprising the amino acid sequence of SEQ ID NO 51 from ammo acid 1 1 to amino acid 30
  • the present invention provides a composition comprising a protein, wherein said protein comprises an amino acid sequence selected from the group consisting of
  • protem (d) the ammo acid sequence encoded by the cDNA insert of clone K640 deposited under accession number ATCC 98026, the protem being substantially free from other mammalian proteins
  • protem comprises the amino acid sequence of SEQ ID NO 51 or the amino acid sequence of SEQ ID NO 51 from ammo acid 1 1 to ammo acid 30
  • the present invention provides a composition comprising an isolated protein encoded by a polynucleotide selected from the group consisting of
  • such polynucleotide comprises the nucleotide sequence of SEQ ID NO 54 from nucleotide 71 to nucleotide 377, the nucleotide sequence of the full length protein coding sequence of clone K39 deposited under accession number ATCC 98026 , or the nucleotide sequence of the mature protein coding sequence of clone K39 deposited under accession number ATCC 98026
  • the polynucleotide encodes the full length or mature protein encoded by the cDNA insert of clone K39 deposited under accession number ATCC 98026
  • such polynucleotide encodes a protein comprising the amino acid sequence of SEQ ID NO 55 from amino acid 62 to amino acid 81
  • the present invention provides a composition comprising a protein, wherein said protein comprises an amino acid sequence selected from the group consisting of
  • the protem comprises the ammo acid sequence of SEQ ID NO 55 or the amino acid sequence of SEQ ID NO 55 from ammo acid 62 to ammo acid 81
  • the present invention provides a composition comprising an isolated protein encoded by a polynucleotide selected from the group consisting of
  • such polynucleotide comp ⁇ ses the nucleotide sequence of SEQ ID NO 57 from nucleotide 194 to nucleotide 423, the nucleotide sequence of the full length protein coding sequence of clone AT319 deposit. ' under accession number ATCC 98026
  • polynucleotide encodes the full length or mature protein encoded by the cDNA insert of clone AT319 deposited under accession number ATCC 98026
  • polynucleotide encodes a protem comprising the amino acid sequence of SEQ ID NO 58 from amino acid 2 to ammo acid 21
  • the present mvention provides a composition comprising a protein, wherem said protein comprises an amino acid sequence selected from the group consisting of
  • protem comprises the ammo acid sequence of SEQ ID NO 58 or the ammo acid sequence of SEQ ID NO 58 from ammo acid 2 to ammo acid 21
  • Protein compositions of the present invention may further comprise a pharmaceutically acceptable carrier Compositions comprising an antibody which specifically reacts with such protein are also provided by the present invention
  • Methods are also provided for preventing, treating or ameliorating a medical condition which comprises administering to a mammalian subject a therapeutically effective amount of a composition comprising a protein of the present invention and a pharmaceutically acceptable carrier
  • Fig 1 is an autoradiograph evidencing the expression of clones AP162, AM931 , and AR260 in COS cells (expressed band(s) indicated by dot(s))
  • Fig 2 is an autoradiographevidencingthe expressionofclone AM610 in COS cells
  • Fig 3 is an autoradiograph evidencing the expression of clones AM340, AM282 and AK533 in COS cells (expressed band(s) indicated by dot(s))
  • Fig 4 is an autoradiograph evidencing the expression of clone AK647 in COS cells (expressed band(s) indicated by dot(s))
  • Fig 5 is an autoradiograph evidencing the expression of clones AH583, AK296, and AS32 in COS cells (expressed band(s) indicated by dot(s))
  • Fig 6 is an autoradiograph evidencing the expression of clones H617 and AT205 in COS cells (expressed band(s) indicated by dot(s))
  • Fig 7 is an autoradiograph evidencing the expression of clones BB9 and K39 in COS cells (expressed band(s) indicated by dot(s))
  • Fig 8 is an autoradiograph evidencing the expression of clones AW 191 and AS34 in COS cells (expressed band(s) indicated by dot(s))
  • Fig 9 is an autoradiograph evidencing the expression of clones AT21 1 and AT319 in COS cells (expressed band(s) indicated by dot(s))
  • Fig 10 is an autoradiograph evidencing the expression of clone K640 in COS cells (expressed band(s) indicated by dot(s))
  • nucleotide and ammo acid sequences are reported below for each clone and protem disclosed in the present application In some instances the sequences are preliminary and may include some incorrect or ambiguous bases or amino acids
  • the actual nucleotide sequence of each clone can readily be determined by sequencing of the deposited clone m accordance with known methods The predicted ammo acid sequence (both full length and mature) can then be determined from such nucleotide sequence
  • the amino acid sequence of the protein encoded by a particular clone can also be determined by expression of the clone in a suitable host cell, collecting the protein and determining its sequence
  • reported protem sequences include "Xaa” designators These "Xaa” designators indicate either (1) a residue which cannot be identified because of nucleotide sequence ambiguity or (2) a stop codon in the determined nucleotide sequence where applicants believe one should not exist (if the nucleotide sequence were determined definitively)
  • 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 which are transported across the membrane of the endoplpasmic reticulum
  • a partial cDNA clone encoding AP162 was first isolated from a human adult placenta cDNA library using methods which are selective for cDNAs encoding secreted proteins
  • the nucleotide sequence of such partial cDNA was determined and searched against the GenBank database using BLASTA/BLASTX and FASTA search protocols
  • the search revealed at least some identity with an EST reported by the I M A G E Consortium identified as "yu40d08 rl Homo sapiens cDNA clone 23671 5' " (GenBank accession number H62096)
  • the search also found a hit at GenBank accession number H98192
  • the human cDNA clone corresponding to the EST database entry was ordered from Genome Systems, Inc , St Louis, Mo, a distributor of the I M A G E Consonium library
  • the clone received from the distributor was examined and determined to be a full length clone, including a 5' end and 3'
  • Protein "AM931" One protein of the present invention has been identified as protein "AM931 "
  • a partial cDNA clone encoding AM931 was first isolated from a human fetal kidney cDNA library using methods which are selective for cDNAs encoding secreted proteins
  • the nucleotide sequence of such partial cDNA was determined and searched against the GenBank database using BLASTA/BLASTX and FASTA search protocols
  • the search revealed at least some identity with an EST reported by the I M A G E Consortium identified as "yh63e02 rl Homo sapeins cDNA clone 134426 5' " (GenBank accession number R32076)
  • the search also found a hit at GenBank accession number N30331
  • the human cDNA clone corresponding to the EST database entry was ordered from Genome Systems, Inc , St Louis, Mo, a distributor of the I M A G E Consonium library
  • the clone received from the distributor was examined and determined to be
  • AM610' A partial cDNA clone encoding AM610 was first isolated from a human fetal kidney cDNA library using methods which are selective for cDNAs encoding secreted proteins The nucleotide sequence of such partial cDNA was determined and searched against the GenBank database using BLASTA/BLASTX and FASTA search protocols The search revealed at least some identity with an EST reported by the I M A G E Consortium identified as "ymOlalO rl Human EST 46249 5" * (GenBank accession number H09925) The search also found hits at GenBank accession numbers H09926 and R 14298 The human cDNA clone corresponding to the EST database entry was ordered from Genome Systems, Inc., St Louis, Mo, a distributor of the I M A G E Consonium library The clone received from the distributor was examined and determined to be a full length clone, including a 5' end and 3' UTR (including a polyA
  • Ammo acids 1 to 23 are the predicted leader/signal sequence, with the predicted mature amino acid sequence beginning at ammo acid 24 Additional nucleotide sequence from the 3 ' portion of AM610, including the polyA tail, is reported in SEQ ID NO 8
  • Protein "AM340” One protein of the present invention has been identified as protein "AM340"
  • a partial cDNA clone encoding AM340 was first isolated from a human fetal kidney cDNA library using methods which are selective for cDNAs encoding secreted proteins
  • the nucleotide sequence of such partial cDNA was determined and searched against the GenBank database using BLASTA/BLASTX and FASTA search protocols
  • the search revealed at least some identity with an EST reported by the I M A G E Consortium identified as "yo68a05 rl Homo sapiens cDNA clone 183056 5' " (GenBank accession number H42936)
  • the search also found a hit at GenBank accession number H42872
  • the human cDNA clone corresponding to the EST database entry was ordered from Genome Systems, Inc , St Louis, Mo, a distributor of the I M A G E Consortium library
  • the clone received from the distributor was examined and determined to be a full length
  • AM282 A partial cDNA clone encoding AM282 was first isolated from a human fetal kidney cDNA library using methods which are selective for cDNAs encoding secreted proteins The nucleotide sequence of such partial cDNA was determined and searched against the GenBank database using BLASTA/BLASTX and FASTA search protocols The search revealed at least some identity with an EST reported by the I M A G E Consortium identified as "yf95bl0 rl Human EST 30142 5' " (GenBank accession number R18560) The search also found a thiat GenBank accession number T96696 The human cDNA clone corresponding to the EST database entry was ordered from Genome Systems, Inc , St Louis, Mo, a distributor ot the I M A G E Consortium library The clone received from the distributor was examined and determined to be a full length clone, including a 5' end and 3' UTR (including a polyA tail).
  • AK647 A partial cDNA clone encoding AK647 was first isolated from a human fetal kidney cDNA library using methods which are selective for cDNAs encoding secreted proteins The nucleotide sequence of such partial cDNA was determined and searched against the GenBank database using BLASTA/BLASTX and FASTA search protocols The search revealed at least some identity with an EST reported by the I M A G E Consortium identified as "ym40a05 rl Human EST 50483 5' " (GenBank accession number H 17726) The human cDNA clone corresponding to the EST database entry was ordered from Genome Systems, Inc , St Louis, Mo, a distributor of the I M A G E Consortium library The clone received from the distributor was examined and determined to be a full length clone, including a 5' end and 3' UTR (including a polyA tail) This full-length clone is also referred to herein as "
  • Applicants' methods identified clone AK647 as encoding a secreted protein
  • the nucleotide sequence of the 5' portion of AK647 as presently determined is reported in SEQ ID NO 14 What applicants believe is the proper reading frame and the predicted N-terminal ammo acid sequence of the full length AK647 protein corresponding to the foregoing nucleotide sequence is reported in SEQ ID NO 15
  • Ammo acids 1 to 25 are the predicted leader/signal sequence, with the predicted mature ammo acid sequence beginning at amino acid 26
  • Additional nucleotide sequence from the 3 portion of AK647, including the polyA tail is reported in SEQ ID NO 16
  • AK583 A partial cDNA clone encoding AK583 was first isolated from a human fetal kidney cDNA library using methods which are selective for cDNAs encoding secreted proteins The nucleotide sequence of such partial cDNA was determined and searched against the GenBank database using BLASTA/BLASTX and FASTA search protocols The search revealed at least some identity with an EST reported by the I M A G E Consortium identified as ⁇ y ⁇ 90c06 rl Human EST 14656 5"' (GenBank accession number R77830) The search also found a hit at GenBank accession number H45398 The human cDN A clone corresponding to the EST database entry was ordered from Genome Systems, Inc , St Louis, Mo, a distributor of the I M A G E Consortium library The clone received from the distributor was examined and determined to be a full length clone, including a 5' end and 3' UTR (including a poly
  • Applicants' methods identified clone AK583 as encoding a secreted protein
  • the nucleotide sequence of the 5' portion of AK583 as presently determined is reported in SEQ ID NO 17 What applicants believe is the proper reading frame and the predicted N-terminal ammo acid sequence of the full length AK583 protein corresponding to the foregoing nucleotide sequence is reported in SEQ ID NO 18
  • Ammo acids 1 to 24 are the predicted leader/signal sequence, with the predicted mature ammo acid sequence beginning at ammo acid 25
  • Additional nucleotide sequence from the 3' portion of AK583, including the polyA tail is reported in SEQ ID NO 19
  • AK533 A partial cDNA clone encoding AK533 was first isolated from a human fetal kidney cDN A library using methods which are selective for cDNAs encoding secreted proteins The nucleotide sequence of such partial cDNA was determined and searched against the GenBank database using BLASTA/BLASTX and FASTA search protocols The search revealed at least some identity with an EST reported by the I M A G E Consortium identified as "yb82h07 rl Homo sapiens cDNA clone 77725 5' " (GenBank accession number T55939) The human cDNA clone corresponding to the EST database entry was ordered from Genome Systems, Inc , St Louis, Mo, a distributor of the I M A G E Consortium library The clone received from the distributor was examined and determined to be a full length clone, including a 5' end and 3' UTR (including a polyA tail) This full- length clone
  • Applicants' methods identified clone AK533 as encoding a secreted protein
  • the nucleotide sequence of the 5' portion of AK533 as presently determined is reported m SEQ ID NO 20
  • SEQ ID NO 21 Additional nucleotide sequence from the 3' portion of AK533, including the polyA tail, is reported in SEQ ID NO 22
  • AK296 A partial cDNA clone encoding AK296 was first isolated from a human fetal kidney cDNA library using methods which are selective for cDNAs encoding secreted proteins The nucleotide sequence of such partial cDNA was determined and searched against the GenBank database using BLASTA/BLASTX and FASTA search protocols The search revealed at least some identity with an EST reported by the I M A G E Consortium identified as "yc86gl2 rl Homo sapeins cDNA clone 22958 5' " (GenBank accession number T75226) The human cDNA clone corresponding to the EST database entry was ordered from Genome Systems, Inc , St Louis, Mo, a distributor of the I M A G E Consortium library The clone received from the distributor was examined and determined to be a full length clone, including a 5' end and 3' UTR (including a polyA tail) This full- length clon
  • nucleotide sequence of the 5 portion of AK296 as presently determined is reported in SEQ ID NO 23 What applicants believe is the proper reading frame and the predicted N-terminal amino acid sequence of the full length AK296 protein corresponding to the foregoing nucleotide sequence is reported in SEQ ID NO 24
  • Ammo acids 1 to 36 are the predicted leader/signal sequence, with the predicted mature amino acid sequence beginning at ammo acid 37
  • Additional nucleotide sequence from the 3 ' portion of AK296, including the polyA tail, is reported m SEQ ID NO 25
  • H617 A partial cDNA clone encoding H617 was first isolated from a human PBMC cDNA library using methods which are selective for cDNAs encoding secreted proteins
  • the nucleotide sequence of such partial cDN A was determined and searched against the GenBank database using BLASTA/BLASTX and FASTA search protocols
  • the search revealed at least some identity with an EST reported by the I M A G E Consortium identified as "ysl lcl2 rl Homo sapeins cDNA clone 214486 5' " (GenBank accession number H71514)
  • the search also found a hit at GenBank accession number R10010
  • the human cDNA clone corresponding to the EST database entry was ordered from Genome Systems, Inc , St
  • H617 Applicants' methods identified clone H617 as encoding a secreted protein
  • nucleotide sequence of the 5' portion of H617 as presently determined is reported in SEQ ID NO 26 What applicants believe is the proper reading frame and the predicted N-terminal amino acid sequence of the full length H617 protein corresponding to the foregoing nucleotide sequence is reported in SEQ ID NO 27 Additional nucleotide sequence from the 3' portion of H617, including the polyA tail, is reported in SEQ ID NO 26
  • BB9 protein "BB9"
  • BB9 A partial cDNA clone encoding BB9 was first isolated from a human PBMC (TH 1 or Th2) cDNA library using methods which are selective for cDNAs encoding secreted proteins
  • the nucleotide sequence of such partial cDNA was determined and searched against the GenBank database using BLASTA/BLASTX and FASTA search protocols
  • the search revealed at least some identity with an EST reported by the I M A G E Consortium identified as "yd68g04 rl Human cDNA clone 113430 5' " (GenBank accession number T78562)
  • the search also found a thi at GenBank accession number R54388
  • the human cDNA clone corresponding to the EST database entry was ordered from Genome Systems,
  • clone BB9 a full length clone, including a 5 end and 3' UTR (including a polyA tail)
  • This full-length clone is also referred to herein as "BB9" Applicants' methods identified clone BB9 as encoding a secreted protein
  • nucleotide sequence of the 5' portion of BB9 as presently determined is reported in SEQ ID NO 29 What applicants believe is the proper readmg frame and the predicted N-terminal amino acid sequence of the full length BB9 protein corresponding to the foregoing nucleotide sequence is reported in SEQ ID NO 30 Additional nucleotide sequence from the 3' portion of BB9, including the polyA tail, is reported in SEQ ID NO 29
  • a partial cDNA clone encoding AW 191 was first isolated from a human ovary (PA-1 teratocarcmoma) cDNA library using methods which are selective for cDNAs encoding secreted proteins
  • the nucleotide sequence of such partial cDNA was determined and searched against the GenBank database using BLASTA/BLASTX and FASTA search protocols
  • the search revealed at least some identity with an EST reported by the I M.A G E Consortium identified as "ym03dl0 rl Homo sapiens cDNA clone 46942 5' " (GenBank accession number H 10314
  • the search also found a hit at GenBank accession number H05460
  • the human cDN A clone corresponding to the EST database entry was ordered from Genome Systems, Inc , St Louis, Mo, a distributor of the I M A G E Consortium library
  • the clone received from the distributor was examined and determined to be a full length
  • nucleotide sequence of the 5' portion of AW 191 as presently determined is reported in SEQ ID NO 32 What applicants believe is the proper reading frame and the predicted N-terminal amino acid sequence of the full length AW 191 protein corresponding to the foregoing nucleotide sequence is reported m SEQ ID NO 33
  • Ammo acids 1 to 18 are the predicted leader/signal sequence, with the predicted mature amino acid sequence beginning at ammo acid 19
  • a partial cDNA clone encoding AT2I 1 was first isolated from a human lymphocyte and dendritic cell cDNA library using methods which are selective for cDNAs encoding secreted proteins
  • the nucleotide sequence of such partial cDNA was determined and searched against the GenBank database using BLASTA/BLASTX and FASTA search protocols
  • the search revealed at least some identity with an EST reported by the I M A G.E Consortium identified as "yq36f01 rl Homo sapiens cDNA clone 197881 5'" (GenBank accession number R96278)
  • the search also found a hit at GenBank accession number R56077
  • the human cDNA clone corresponding to the EST database entry was ordered from Genome Systems, Inc , St Louis, Mo, a distributor of the I M A G E Consortium library
  • the clone received from the distributor was examined and determined to be a full length clone, including a 5' end and
  • AT205 A partial cDNA clone encoding AT205 was first isolated from a human lymphocyte and dendritic cell cDNA library using methods which are selective for cDNAs encoding secreted proteins
  • the nucleotide sequence of such partial cDNA was determined and searched against the GenBank database using BLASTA/BLASTX and FASTA search protocols
  • the search revealed at least some identity with an EST reported by the I M A G E Consortium identified as 'yu83c l 1 rl Homo sapiens cDNA clone 240404 5" (GenBank accession number H78080)
  • the search also found a hit at GenBank accession number H78081
  • the human cDN A clone corresponding to the EST database entry was ordered from Genome Systems, Inc , St Louis, Mo, a distributor of the I M A G E Consortium library
  • the clone received from the distributor was examined and determined to be a full length clone, including a 5' end
  • AS34 A partial cDNA clone encoding AS34 was first isolated from a human fetal brain cDNA library using methods which are selective for cDNAs encoding secreted proteins
  • the nucleotide sequence of such partial cDNA was determined and searched against the GenBank database using BLASTA/BLASTX and FASTA search protocols
  • the search revealed at least some identity with an EST reported by the I M A G E Consortium identified as "yg71a01 rl Homo sapiens cDNA clone 38531 5' " (GenBank accession number R51 1 18)
  • the search also found a hit at GenBank accession number R 15801
  • the human cDNA clone corresponding to the EST database entry was ordered from Genome Systems, Inc , St Louis, Mo, a distributor of the I M A G E Consortium library
  • the clone received from the distributor was examined and determined to be a full length clone, including a 5' end and 3' UTR (
  • nucleotide sequence of the 5' portion of AS34 as presently determined is reported in SEQ ID NO 40 What applicants believe is the proper reading frame and the predicted N-terminal amino acid sequence of the full length AS34 protein corresponding to the foregoing nucleotide sequence is reported in SEQ ID NO 41
  • Ammo acids 1 to 24 are the predicted leader/signal sequence, with the predicted mature amino acid sequence beginning at ammo acid 25
  • AS32 A partial cDNA clone encoding AS32 was first isolated from a human fetal brain cDNA library using methods which are selective for cDNAs encoding secreted proteins
  • the nucleotide sequence of such partial cDNA was determined and searched against the GenBank database using BLASTA/BLASTX and FASTA search protocols
  • the search revealed at least some identity with an EST reported by the I M A G E Consortium identified as "yu75b08 rl Homo sapiens cDNA clone 239607 5' " (GenBank accession number H80466)
  • the search also found a hit at GenBank accession number H77627
  • the human cDNA clone corresponding to the EST database entry was ordered from Genome Systems, Inc , St Louis, Mo, a distributor of the I M A G E Consortium library
  • the clone received from the distributor was examined and determined to be a full length clone, including a 5' end and 3' UTR (including
  • One protein of the present invention has been identified as protein ⁇ R260"
  • a partial cDNA clone encoding AR260 was first isolated from a human adult retina cDNA library using methods which are selective for cDNAs encoding secreted proteins The nucleotide sequence of such partial cDNA was determined and searched against the
  • GenBank database using BLASTA/BLASTX and FASTA search protocols The search revealed at least some identity with an EST reported by the I M A G E Consortium identified as "yg99gl2 rl Homo sapiens cDNA clone 41757 5' " (GenBank accession number R52804)
  • the human cDNA clone corresponding to the EST database entry was ordered from Genome Systems, Inc , St Louis, Mo, a distributor of the I M A G E
  • Applicants' methods identified clone AR260 as encoding a secreted protein
  • the nucleotide sequence of the 5 portion of AR260 as presently determined is reported in SEQ ID NO 46 What applicants believe is the proper reading frame and the predicted N-terminal ammo acid sequence of the full length AR260 protein corresponding to the foregoing nucleotide sequence is reported in SEQ ID NO 47
  • Amino acids 1 to 23 are the predicted leader/signal sequence, with the predicted mature ammo acid sequence beginning at ammo acid 24
  • Additional nucleotide sequence from the 3 portion of AR260, mcluding the polyA tail is reported in SEQ ID NO 48
  • K640 A partial cDNA clone encoding K640 was first isolated from a murine bone marrow (stromal cell line FCM-4) cDNA library using methods which are selective for cDNAs encoding secreted proteins
  • the nucleotide sequence of such partial cDN A was determined and searched against the GenBank database using BLASTA/BLASTX and FASTA search protocols
  • the search revealed at least some identity with an EST reported by the I M A G E Consortium identified as 'yf47a09 rl Homo sapiens cDNA clone 129976 5' " (GenBank accession number Rl 1595)
  • the search also found a hit at GenBank accession number H09031
  • the human cDNA clone corresponding to the EST database entry was ordered from Genome Systems, Inc , St Louis, Mo, a distributor of the I M A G E Consortium library
  • the clone received from the distributor was examined and determined to be a full length clone
  • nucleotide sequence of the 5' portion of K640 as presently determined is reported in SEQ ID NO 49 What applicants believe is the proper reading frame and the predicted N-terminal amino acid sequence of the full length K640 protein corresponding to the foregoing nucleotide sequence is reported in SEQ ID NO 50 Additional nucleotide sequence from the 3' portion of K640, including the polyA tail, is reported in SEQ'ID
  • K39 A partial cDNA clone encoding K39 was first isolated from a murine bone marrow (stromal cell line FCM-4) cDNA library using methods which are selective for cDNAs encoding secreted proteins. The nucleotide sequence of such partial cDNA was determined and searched against the GenBank database using BLASTA/BLASTX and FASTA search protocols. The starch revealed at least some identity with an EST reported by the I.M.A.G.E. Consortium identified as "ym65b04.rl Homo sapiens cDNA clone 163759 5' " (GenBank accession number H 14129). The search also found a hit at GenBank accession number H68304.
  • the human cDN A clone corresponding to the EST database entry was ordered from Genome Systems, Inc. , St. Louis, Mo, a distributor of the I.M.A.G.E. Consortium library.
  • the clone received from the distributor was examined and determined to be a full length clone, including a 5' end and 3' UTR (including a polyA tail).
  • This full-length clone is also referred to herein as "K39" .
  • Applicants' methods identified clone K39 as encoding a secreted protein.
  • nucleotide sequence of the 5' portion of K39 as presently determined is reported in SEQ ID NO:52. What applicants believe is the proper reading frame and the predicted N-terminal amino acid sequence of the full length K39 protein corresponding to the foregoing nucleotide sequence is reported in SEQ ID NO:53. Additional nucleotide sequence from the 3' portion of K39, including the polyA tail, is reported in SEQ ID NO:54.
  • AT319 A partial cDNA clone encoding AT319 was first isolated from a human lymphocyte and dendritic cell cDNA library using methods which are selective for cDNAs encoding secreted proteins .
  • the nucleotide sequence of such partial cDN A was determined and searched against the GenBank database using BLASTA/BLASTX and FASTA search protocols. The search revealed at least some identity with an EST reported by the I.M.A.G.E. Consortium identified as "yr21bl l .rl Homo sapiens cDNA clone 205917 5'" (GenBank accession number H57730). The search also found a hit at GenBank accession number H57731.
  • the human cDN A clone corresponding to the EST database entry was ordered from Genome Systems, Inc , St Louis. Mo, a distributor of the I M A.G E Consortium library
  • the clone received from the distributor was examined and determined to be a full length clone, including a 5' end and 3' UTR (including a polyA tail)
  • This full- length clone is also referred to herein as "AT319" Applicants' methods identified clone AT319 as encoding a secreted protein
  • nucleotide sequence of the 5' portion of AT319 as presently determined is Reported in SEQ ID N0 55 What applicants believe is the proper reading frame and the predicted N-terminai amino acid sequence of the full length AT319 protein corresponding to the foregoing nucleotide sequence is reported in SEQ ID NO:56 Additional nucleotide sequence from the 3' portion of AT319, including the polyA tail, is reported in SEQ ID NO:57
  • Clones AP162, AM931 , AM610, AM340, AM282, AK647. AK583. AK533, AK296, H617, BB9, AW191 , AT211 , AT205, AS34, AS32, AR260, K640, K39 and AT319 were deposited on April 17, 1996 with the American Type Culture Collection under accession number ATCC 98026, from which each clone comprising a particular polynucleotide is obtainable Each clone has been transfected into separate bacterial cells (E colt) m this composite deposit Bacterial cells containing a particular clone can be obtained from the composite deposit as follows
  • 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 design of the oligonucleotide probe should preferably follow these parameters (a) It should be designed to an area of the sequence which has the fewest ambiguous bases ("N's"), if any, (b) It should be designed to have a T m of approx 80 ° C (assuming 2° for each
  • the oligonucleotide should preferably be labeled with g- 32 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 inco ⁇ orated into the probe should be quantitated by measurement in a scintillation counter Preferably, specific activity of the resulting probe should be approximately 4e + 6 dpm/pmole
  • the bacterial culture containing the pool of full-length clones should preferably be thawed and 100 ⁇ l of the stock used to inoculate a sterile culture flask containing 25 ml of sterile L-broth containing ampicillin at 100 ⁇ g/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 ⁇ g/ml and agar at 1 5% in a 150 mm petri dish when grown overnight at 37°C Other known methods of obtaining distinct, well-separated colonies can also be employed
  • Standard colony hybridization procedures should then be used to transfer the colonies to nitrocellulose filters and lyse, denature and bake them
  • the filter is then preferably incubated at 65 °C for 1 hour with gentle agitation in 6X SSC (20X stock is 175.3 g NaCI/liter, 88.2 g Na citrate/liter, adjusted to pH 7.0 with NaOH) containing 0.5% SDS, 100 ⁇ g/ml of yeast RNA, and 10 mM EDTA (approximately 10 mL per 150 mm filter).
  • 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 0.1X SSC/0.5 % SDS at 65 °C for 30 minutes to 1 hour is optional.
  • the filter is then preferably dried and subjected to autoradiography for sufficient time to visualize the positives on the X-ray film. Other known hybridization methods can also be employed.
  • the positive colonies are picked, grown in culture, and plasmid DNA isolated using standard procedures.
  • the clones can then be verified by restriction analysis, hybridization analysis, or DNA sequencing.
  • Fragments of the proteins of the present invention which are capable of exhibiting biological activity are also encompassed by the present invention.
  • Fragments of the protein may be in linear form or they may be cyclized using known methods, for example, as described in H.U. Saragovi, et al. , Bio/Technology 10, 773-778 (1992) and in R.S. McDowell, et al. , J. Amer. Chem. Soc. J_14, 9245-9253 ( 1992), both of which are inco ⁇ orated herein by reference.
  • Such fragments may be fused to carrier molecules such as immunoglobulins for many pu ⁇ oses, including increasing the valency of protein binding sites.
  • fragments of the protein may be fused through "linker" sequences to the Fc portion of an immunoglobulin.
  • 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.
  • 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 form 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 sequence of the mature form of the protein may also be determinable from the amino acid sequence of the full-length form.
  • the present invention also provides for soluble forms of such protein.
  • pan or ail of the intracellular and transmembrane domains of the protein are deleted such that the protein is fully secreted from the cell in which it is expressed.
  • the intracellular and fransmembrane domains of proteins of the invention can be identified in accordance with known techniques for determination of such domains from sequence information.
  • Species homologs of the disclosed proteins are also provided by the present invention. Species homologs 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.
  • the invention also encompasses allelic variants of the disclosed proteins; that is, naturally-occurring alternative forms of the isolated proteins which are identical, homologous or related to that encoded by the polynucleotides disclosed herein.
  • the isolated polynucleotide endcoing the protein of the invention may be operably linked to an expression control sequence such as the pMT2 or pED expression vectors disclosed in Kaufman et al.. Nucleic Acids Res. __, 4485-4490 (1991), in order to produce the protein recombinantly.
  • an expression control sequence such as the pMT2 or pED expression vectors disclosed in Kaufman et al.. Nucleic Acids Res. __, 4485-4490 (1991)
  • 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 185. 537-566 (1990).
  • operably 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.
  • Mammalian host cells include, for example, monkey COS cells, Chinese Hamster Ovary (CHO) cells, human kidney 293 cells, human epidermal A431 cells, human Colo205 cells, 3T3 cells, CV-1 cells, other transformed primate cell lines, normal diploid cells, cell strains derived from in vitro culture of primary tissue, primary explants, HeLa cells, mouse
  • L cells BHK, HL-60, U937, HaK or Jurkat cells.
  • yeast in lower eukaryotes such as yeast or in prokaryotes such as bacteria.
  • yeast strains include Saccharomyces cerevisiae, Schizosaccharomyc ⁇ pombe, Kluyveromyces strains, Candida, or any yeast strain capable of expressing heterologous proteins
  • Potentially suitable bacterial strains include Escherichia colt, Bacillus subtilis, Salmonella rvphtmurium, 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, m order to obtain the functional protein Such covalent attachments may be accomplished using known chemical or enzymatic methods
  • the protein may also be produced by operably linking the isolated polynucleotide of the invention to suitable control sequences in one or more insect expression vectors, and employing an insect expression system
  • suitable control sequences in one or more insect expression vectors
  • an insect expression system Materials and methods for baculovirus/insect cell expression systems are commercially available in kit form from, e g , Invitrogen, San Diego, California, U S A (the MaxBac ® kit), and such methods are well known in the art, as described in Summers and Smith, Texas Agricultural Experiment Station Bulletin No 1555 (1987). inco ⁇ orated herein by reference
  • 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 containing agents which will bind to the protein, one or more column steps over such affinity resins as concanavalin A-agarose, hepa ⁇ n-toyopearl ® or Cibacrom blue 3GA Sepharose ® , one or more steps involving hydrophobic interaction chromatography using such resins as phenyl ether, butyl ether, or propyl ether, or immunoaffinity chromatography
  • the protein of the invention may also be expressed in a form which will facilitate purification
  • it may be expressed as a fusion protein, such as those of maltose binding protein (MBP), glutathione-S-transferase (GST) or thioredoxin (TRX) Kits for expression and purification of such fusion proteins are commercially available from New England BioLab (Beverly, MA), Pharmacia (Piscataway, NJ) and
  • the protein can also be tagged with an epitope and subsequently purified by using a specific antibody directed to such epitope
  • a specific antibody directed to such epitope One such epitope (“Flag") is commercially available from Kodak (New Haven, CT)
  • RP- HPLC reverse-phase high performance liquid chromatography
  • hydrophobic RP-HPLC media e g , sihca gel having pendant methyl or other aliphatic groups
  • 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-constructedprotein sequences by virtue of sharing primary, secondary or tertiary structural and/or conformational characteristics with proteins may possess biological properties in common therewith, including protein activity Thus, they may be employed as biologically active or immunological substitutes for natural, purified proteins in screening of therapeutic compounds and in immunological processes for the development of antibodies
  • the proteins provided herein also include proteins characte ⁇ zed by amino acid sequences similar to those of purified proteins but into which modification are naturally provided or deliberately engineered
  • 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, insertion or deletion of a selected ammo acid residue in the coding sequence
  • 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 alter
  • 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)
  • 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-hgand 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 bmding 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
  • Proteins of the present invention can also be used as nutritional sources or supplements . Such uses include without limitation use as a protein or amino acid supplement, use as a carbon source, use as a nitrogen source and use as a source of carbohydrate.
  • the protein 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.
  • the protein of the invention can be added to the medium in or on which the microorganism is cultured.
  • 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.
  • cytokine cytokine
  • cell proliferation either inducing or inhibiting
  • cell differentiation either inducing or inhibiting
  • 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, B9/1 1 , BaF3, MC9/G, M + (preB M + ), 2E8, RB5, DAI , 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.
  • Assays for cytokine production and/or proliferation of spleen cells, lymph node 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.
  • Assays for prohferationand differentiationof hematopoieticand lymphopoietic cells include, without limitation, those described in Measurement of Human and Murine Interieukin 2 and Interieukin 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 , deV ⁇ es et al , J Exp Med 173 1205-1211, 1991, Moreau et al , Nature 336 690-692, 1988, Greenberger et al , Proc Natl Acad Sci U S A 80 2931-2938, 1983, Measurement of mouse and human interieukin 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
  • a protein of the present invention may also exhibit immune stimulating or immune suppressing activity, including without limitation the activities for which assays are described herein
  • a protein may be useful in the treatment of various immune deficiencies and disorders (including severe combined immunodeficiency (SCID)), e g , in regulating (up or down) growth and proliferation of T and/or B lymphocytes, as well as effecting the cytolytc activity of NK cells and other cell populations
  • SCID severe combined immunodeficiency
  • 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
  • infectious 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, he ⁇ esviruses, mycobacte ⁇ a, Leishmania spp , malaria spp and various fungal infections such as candidiasis
  • a protein of the present invention may also be useful where
  • Autoimmune disorders which may be treated using a protein of the present invention include, for example, connective tissue disease, multiple sclerosis, systemic lupus erythematosus, rheumatoid arthritis, autoimmune pulmonary inflammation, Guillain-Barre syndrome, autoimmune thyroiditis, insulin dependent diabetes melhtis, myasthenia gravis, graft-versus-hostdisease and autoimmune inflammatory eye disease
  • 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 may also be treatable using a protein of the present invention
  • 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 tole ⁇ zing 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 tole ⁇ zing agent Down regulating or preventing one or more antigen functions (including without limitation B lymphocyte antigen functions (such as , for example, B7)), e g , preventing high level lymphokme
  • the efficacy of particular blocking reagents in preventing organ transplant rejection or GVHD can be assessed using animal models that are predictive of efficacy in humans
  • animal models that are predictive of efficacy in humans
  • 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 257789-792 (1992) and Turka et al , Proc Natl.
  • 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 autoantibodes involved in the pathology of the diseases
  • Preventing the 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 m the disease process
  • blocking reagents may induce antigen-specific tolerance of autoreactive T cells which could lead to long-term relief from the disease.
  • the efficacy of blocking reagents in preventing or alleviating autoimmune disorders can be determined using a number of well-characterized animal models of human autoimmune diseases. Examples include murine experimental autoimmune encephalitis, systemic lupus erythmatosis in MRL/lpr/lpr mice or NZB hybrid mice, murine autoimmune collagen arthritis, diabetes mellitus in NOD mice and BB rats, and murine 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), a's a means of up regulating immune responses, may also be useful in therapy. Upregulation of immune responses may be in the form of enhancing an existing immune response or eliciting an initial immune response. For example, enhancing an immune response through stimulating B lymphocyte antigen function may be useful in cases of viral infection. In addition, systemic viral diseases such as influenza, the common cold, and encephalitis might be alleviated by the administration of stimulatory forms of B lymphocyte antigens systemically.
  • anti-viral immune responses may be enhanced in an infected patient by removing T cells from the patient, costimulating the T cells in vitro with viral antigen- pulsed APCs either expressing a peptide of the present invention or together with a stimulatory form of a soluble peptide of the present invention and reintroducing the in vitro activated T cells into the patient.
  • Another method of enhancing anti-viral immune responses would be to isolate infected cells from a patient, transfect them with a nucleic acid encoding a protein of the present invention as described herein such that the cells express all or a portion of the protein on their surface, and reintroduce the transfected cells into the patient.
  • the infected cells would now be capable of delivering a costimulatory signal to, and thereby activate, T cells in vivo.
  • up regulation or enhancement of antigen function may be usefiil in the induction of tumor immunity.
  • Tumor cells e.g., sarcoma, melanoma, lymphoma, leukemia, neuroblastoma, carcinoma
  • 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.
  • the tumor cell can be transfected to express a combination of peptides .
  • tumor cells obtained from a patient can be transfected ex vivo with an expression vector directing the expression of a peptide having B7-2-like activity alone, or in conjunction with a peptide having B7-l-like activity and/or B7-3-like activity.
  • the transfected tumor cells are returned to the patient to result in expression of the peptides on the surface of the transfected cell
  • gene therapy techniques can be used to target a tumor cell for transfection in vivo
  • tumor cells which lack MHC class I or MHC class II molecules, or which fail to reexpress sufficient amounts of MHC class I or MHC class II molecules, can be transfected with nucleic acid encoding all or a portion of (e g , a cytoplasmic-domain truncated portion) of an MHC class I ⁇ chain protein and ⁇ 2 microglobuhn protein or an MHC class II a chain protein and an MHC class II ⁇ 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-l, B7-2, B7-3 induces a T cell surface
  • T-cell-dependent immunoglobulin responses and isotype switching include, without limitation, those described in Mahszewski, 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
  • MLR Mixed lymphocyte reaction
  • 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 m Darzynkiewicz et al , Cytometry 13 795-S08, 1992, Gorczyca et al , Leukemia 7 659-670, 1993, Gorczyca et al , Cancer Research 53 1945-1951 , 1993, Itoh et al , Cell 66 233-243, 1991 , Zacharchuk, Journal of Immunology 145 4037-4045, 1990, Zamai et al , Cytometry 14 891-897, 1993, Gorczyca et al , International Journal of Oncology 1 639-648, 1992
  • a protein of the present invention may be useful in regulation of hematopoiesis and, consequently, in the treatment of myeloid or lymphoid cell deficiencies. Even marginal biological activity in support of colony forming cells or of factor-dependent cell lines indicates involvement in regulating hematopoiesis, e.g.
  • erythroid progenitor cells alone or in combination with other cytokines, (hereby indicating utility, for example, in treating various anemias or for use in conjunction with irradiation/chemotherapy to stimulate the production of erythroid precursors and/or erythroid cells; in supporting the growth and proliferation of myeloid cells such as granulocytes and monocytes/macrophages (i.e., traditional CSF activity) useful, for example, in conjunction with chemotherapy to prevent or treat consequent myelo ⁇ suppression; in supporting the growth and proliferation of megakaryocytesand 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
  • the activity of a protein of the invention may, among other means, be measured by the following methods: Suitable assays for proliferation and differentiation of various hematopoietic lines are cited above.
  • Assays for embryonic stem cell differentiation include, without limitation, those described in: Johansson et al. Cellular Biology 15: 141-151 , 1995; Keller et al., Molecular and Cellular Biology 13:473-486, 1993; McClanahan et al.. Blood 81 :2903-2915, 1993.
  • Assays for stem cell survival and differentiation 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 89:5907-591 1 , 1992; Primitive hematopoietc colony forming cells with high proliferative potential, McNiece, I K. and Briddell, R.A.
  • a protein of the present invention also may have utility in compositions used for bone, cartilage, tendon, ligament and/or nerve tissue growth or regeneration, as well as for wound healing and tissue repair and replacement, and in the treatment of burns, incisions and ulcers.
  • a protein of the present invention which induces cartilage and/or bone growth in circumstances where bone is not normally formed, has application in the healing of bone fractures and cartilage damage or defects in humans and other animals.
  • Such a preparation employing a protein of the invention may have prophylactic use in closed as well as open fracture reduction and also in the improved fixation of artificial joints. De novo bone formation induced by an osteogenic agent contributes to the repair of congenital, trauma induced, or oncologic resection induced craniofacial defects, and also is useful in cosmetic plastic surgery.
  • a protein of this invention may also be used in the treatment of periodontal 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 invention which induces tendon/ligament-like tissue or other tissue formation in circumstances where such tissue is not normally formed, has application in the healing of tendon or ligament tears, deformities and other tendon or ligament defects in humans and other animals.
  • Such a preparation employing a tendon/ligament-liketissue 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.
  • compositions of the present invention contributes to the repair of congenital, trauma induced, or other tendon or ligament defects of other origin, and is also useful in cosmetic plastic surgery for attachment or repair of tendons or ligaments.
  • the compositions of the present invention may provide an environment to attract tendon- or ligament-forming cells, stimulate growth of tendon- or ligament-forming cells, induce differential ion 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, ca ⁇ al tunnel syndrome and other tendon or ligament defects.
  • compositions may also include an appropriate matrix and/or sequestering agent as a carrier as is well known in the art.
  • the protein of the present invention may also be useful for proliferation of neural cells and for regeneration of nerve and brain tissue, i.e. for the treatment of central and peripheral nervous system diseases and neuropathies, as well as mechanical and traumatic disorders , which involve degeneration, death or trauma to neural cells or nerve tissue. More specifically, a protein may be used in the treatment of diseases of the peripheral nervous system, such as peripheral nerve injuries, peripheral neuropathy and localized neuropathies, and central nervous system diseases, such as Alzheimer's, 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 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
  • 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) ahd vascular (including vascular endothelium) tissue, or for promoting the growth of cells comprising such tissues Part of the desired effects may be by inhibition or modulation of fibrotic scarring to allow normal tissue to regenerate A protein of the invention may also exhibit angiogenic activity
  • a protein of the present invention may also be useful for gut protection or regeneration and treatment of lung or liver fibrosis, reperfusion injury in various tissues, and conditions resulting from systemic cytokine damage
  • a protein of the present invention may also be useful for promoting or inhibiting differentiation of tissues described above from precursor tissues or cells, or for inhibiting the growth of tissues described above
  • the activity of a protein of the invention may, among other means, be measured by the following methods
  • Assays for tissue generation activity include, without limitation, those described in International Patent Publication No W095/ 16035 (bone, cartilage, tendon), International Patent Publication No WO95/05846 (nerve, neuronal), International Patent Publication No W091/07491 (skin, endothelium )
  • Assays for wound healing activity include, without limitation, those described in Winter, Epidermal Wound Healing, pps 71-1 12 (Maibach, HI and Rovee, DT, eds ), Year Book Medical Publishers, Inc , Chicago, as modified by Eaglstein and Mertz, J Invest
  • 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 activms and are characterized by their ability to stimulate the release of follicle stimulating hormone (FSH)
  • FSH follicle stimulating hormone
  • a protein of the present mventioa alone or in heterodimers with a member of the inhibin ⁇ 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 .
  • the protein of the invention 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 dnset of fertility in sexually immature mammals, so as to increase the lifetime reproductive performance of domestic animals such as cows, sheep and pigs.
  • the activity of a protein of the invention may, among other means, be measured by the following methods:
  • Assays for activin/inhibin activity include, without limitation, those described in: Vale et al. , Endocrinology 91 :562-572, 1972; Ling et al.. Nature 321 :779-782, 1986; Vale et al., Nature 321 :776-779, 1986; Mason et al.. Nature 318:659-663, 1985; Forage et al., Proc. Natl. Acad. Sci. USA 83:3091-3095, 1986.
  • a protein of the present invention may have chemotactic or chemokinetic activity (e.g., act as a chemokine) for mammalian cells, including, for example, monocytes, fibroblasts, neutrophils, T-cells, mast cells, eosinophils, epithelial and/or endothelial cells.
  • Chemotactic and chemokinetic proteins can be used to mobilize or attract a desired cell population to a desired site of action.
  • Chemotactic or chemokinetic proteins provide particular advantages in treatment of wounds and other trauma to tissues, as well as in treatment of localized infections. For example, attraction of lymphocytes, monocytes or neutrophils to tumors or sites of infection may result in improved immune responses against the tumor or infecting agent.
  • a protein or peptide has chemotactic activity for a particular cell population if it can stimulate, directly or indirectly, the directed orientation or movement of such cell population.
  • 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 as ⁇ ay for cell chemotaxis.
  • the activity of a protein of the invention may, among other means, be measured 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 include, without limitation, those described in: Current Protocols in Immunology, Ed by J.E. Coligan, A.M. Kruisbeek, D.H. Marguiies, E.M. Shevach, W. Strober, Pub.
  • 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.
  • a protein of the present invention may also demonstrate activity as receptors, receptor ligands or inhibitors or agonists of receptor/ligand interactions.
  • receptors and ligands include, without limitation, cytokine receptors and their ligands, receptor kinases and their ligands, receptor phosphatases and their ligands, 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 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. Marguiies, 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: 1 145-1 156, 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.
  • 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 inflammatory response syndrome (SIRS)), ischemia-reperfusioninjury, 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.
  • infection such as septic shock, sepsis or systemic inflammatory response syndrome (SIRS)
  • ischemia-reperfusioninjury endotoxin lethality
  • arthritis complement-mediated hyperacute rejection
  • nephritis cytokine or chemokine- induced lung injury
  • inflammatory bowel disease Crohn's
  • 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 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 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.
  • 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 or body part size or shape (such as, for example, breast augmentation or diminution, change in bone form or shape); effecting biorhythms or caricadic cycles or rhythms; effecting the fertility of male or female subjects; effecting the metabolism, catabolism, anabolism, processing, utilization, storage or elimination of dietary fat, lipid, protein, carbohydrate, vitamins, minerals, cofactors or other nutritional factors or component(s); effecting behavioral characteristics, including, without limitation, appetite, libido, stress, cognition (including cognitive disorders), depression (including depressive disorders) and violent behaviors; providing analgesic effects or other pain reducing effects; promoting differentiation
  • a protein of the present invention may be used in a pharmaceutical composition when combined with a pharmaceutically acceptable carrier.
  • a pharmaceutically acceptable carrier may also contain (in addition to protein and a carrier) diluents, fillers, salts, buffers, stabilizers, solubilizers, and other materials well known in the art.
  • pharmaceutically acceptable means a non-toxic material that does not interfere with the effectiveness of the biological activity of the active ingredient(s). The characteristics of the carrier will depend on the route of administration.
  • the pharmaceutical composition of the invention may also contain cytokines, lymphokines, or other hematopoietic factors such as M-CSF, GM-CSF, TNF, IL-1 , IL-2, IL-3, IL-4, IL-5, IL-6, IL-7, IL-8, IL-9, IL-10, IL-11, IL-12, IL-13, IL-14, IL-15, IFN, TNFO, TNF1 , TNF2, G-CSF, Meg-CSF, thrombopoietin, stem cell factor, and erythropoietin.
  • the pharmaceutical composition may further contain other agents which either enhance the activity of the protein or compliment its activity or use in treatment.
  • protein of the present invention may be included in formulations of the particular cytokine, lymphokine, other hematopoietic factor, thrombolytic or anti-thrombotic factor, or anti-inflammaory agent to minimize side effects of the cytokine, lymphokine, other hematopoietic factor, thrombolytic or anti-thrombotic factor, or anti-inflammatory agent.
  • a protein of the present invention may be active in multimers (e.g., heterodimers or homodimers) or complexes with itself or other proteins.
  • 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 protein(s) 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.
  • TCR T cell receptor
  • 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 antigen(s) 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.
  • antibodies able to bind surface immunolgobulin and other molecules on B cells as well as antibodies able to bind the TCR and other molecules on T cells can be combined with the pharmaceutical composition of the invention
  • the pharmaceutical composition of the invention may be in the form of a liposome in which protein of the present invention is combined, in addition to other pharmaceutically acceptable carriers, with amphipathic agents such as lipids which exist in aggregated form as micelles, insoluble monolayers, liquid crystals, or lamellar layers m aqueous solution
  • amphipathic agents such as lipids which exist in aggregated form as micelles, insoluble monolayers, liquid crystals, or lamellar layers m aqueous solution
  • Suitable lipids for liposomal formulation include, without limitation, monoglycerides, diglyce ⁇ des, sulfatides, lysolecithin, phospholipids, sapomn, bile acids, and the like
  • Preparation of such liposomal formulations is within the level of skill in the art, as disclosed, for example, m 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
  • the term "therapeutically effective amount” means the total amount of each active component of the pharmaceutical composition or method that is sufficient to show a meaningful patient benefit, l 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
  • the term refers to that ingredient alone
  • the term refers to combined amounts of the active ingredients that result in the therapeutic effect, whether administered in combination, serially or simultaneously
  • a therapeutically effective amount of protein of the present invention is administered to a mammal having a condition to be treated
  • Protein of the present invention may be administered in accordance with the method of the invention either alone or in combination with other therapies such as treatments employing cytokines, lymphokines or other hematopoietic factors
  • protein of the present invention may be administered either simultaneously with the cytokme(s), lymphokme(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 m combination with cytokme(s), lymphok ⁇ ne(s), other hematopoietic factor(s), thrombolytic or anti-thrombotic factors
  • Administration of protein of the present invention used in the pharmaceutical composition or to practice the method of the present invention can be carried out in a variety of conventional ways, such as oral ingestion, inhalation, topical application or cutaneous, subcutaneous, intraperitoneal, parenteral or intravenous injection Intravenous administration to the patient is preferred
  • protein of the present invention When administered orally, protein of the present invention will be in the form of a tablet, capsule, powder, solution or elixir
  • the pharmaceutical cOmposition of the invention may additionally contain a solid carrier such as a gelatin or an adjuvant
  • the tablet, capsule, and powder contain from about 5 to 95% protein of the present invention, and preferably from about 25 to 90% protein of the present invention
  • 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 physiological saline solution, dextrose or other saccharide solution, or glycols such as ethylene glycol, propylene glycol or polyethylene glycol
  • 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
  • protein of the present invention When a therapeutically effective amount of protein of the present invention is administered by intravenous, cutaneous or subcutaneous injection, protein of the present invention will be in the form of a pyrogen-free, parenterally acceptable aqueous solution
  • 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 increased further It is contemplated that the various pharmaceutical compositions used to practice the method of the present invention should contain about 0 01 ⁇ g to about 100 mg (preferably about 0 l ⁇ g to about 10 mg, more preferably about 0 1 ⁇ g to about 1 mg) of protein of the present invention per kg body weight
  • the duration of intravenous therapy using the pharmaceutical composition of the present invention will vary, depending on the severity of the disease being treated and the condition and potential idiosyncratic response of each individual patient It is contemplated that the duration of each application of the protein of the present invention will be in the range of 12 to 24 hours of continuous intravenous administration Ultimately the attending physician will decide on the appropriate duration of intravenous therapy using the pharmaceutical composition of the present invention
  • Protein of the invention may also be used to immunize animals to obtain 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 £5_, 2149-2154 (1963), J L Krstenansky, et al , FEBS Lett 211, 10 (1987) Monoclonal antibodies binding to the protein of the invention may be useful diagnostic agents for the immunodetection of the protein
  • Neutralizing monoclonal antibodies binding to the protein may also be useful therapeutics for both conditions associated with the protein and also in the treatment of some forms of cancer where abnormal expression of the protein is involved In the case of cancerous cells or Ieuk
  • the therapeutic method includes administering 'the composition topically, systematically, or locally as an implant or device
  • the therapeutic composition for use in this invention is, of course, in a pyrogen-free, physiologically acceptable form
  • the composition may desirably be encapsulated or injected in a viscous form for delivery to the site of bone, cartilage or tissue damage
  • Topical administration may be suitable for wound healing and tissue repair
  • Therapeutically useful agents other than a protein of the invention which may also optionally be included m the composition as described above, may alternatively or additionally, be administered simultaneously or sequentially with the composition in the methods of the invention
  • the composition would include a matrix capable of delivering the protein-containing composition to the site df bone and/or cartilage damage, providing a structure for the developing bone and cartilage and optimally capable of being resorbed into the body
  • Such matrices may be formed of materials presently in use for other implant
  • biodegradable and biologically well-defined such as bone or dermal collagen
  • Further matrices are comprised of pure proteins or extracellular matrix components
  • Other potential mat ⁇ ces 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 t ⁇ calciumphosphate
  • 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
  • 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
  • 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(mcludinghydroxyalkylcelluloses), including methylcellulose, ethylcellulose, hydroxyethylcellulose, hydroxypropylcellulose, hydroxypropyl-methylcellulose, &nd carboxymethylcellulose, the most preferred being cationic salts of carboxymethylcellulose (CMC)
  • CMC carboxymethylcellulose
  • Other preferred sequestering agents include hyaluronic acid, sodium alginate, polyethylene glycol), polyoxyethylene oxide, carboxyvinyl polymer and poly(vmyl alcohol)
  • the amount of sequestering agent useful herein is 0 5-20 wt%, preferably 1-10 wt% based on total formulation weight, which represents the amount necessary to prevent desorbtion of the protein from the polymer matrix and to provide appropriate handling of the composition, yet not so much that the progenitor cells are prevented from infiltrating the matrix, thereby providing the protein the opportunity to assist the osteogenic activity of the
  • 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.
  • agents include various growth factors such as epidermal growth factor (EGF), platelet derived growth factor (PDGF), transforming growth factors (TGF- ⁇ and TGF- ⁇ ), and insulin-like growth factor (IGF).
  • EGF epidermal growth factor
  • PDGF platelet derived growth factor
  • TGF- ⁇ and TGF- ⁇ transforming growth factors
  • IGF insulin-like growth factor
  • the therapeutic compositions are also presently valuable for veterinary applications. Particularly domestic animals and thoroughbred horses, in addition to humans, are desired patients for such treatment with proteins of the present invention.
  • the dosage regimen of a protein-containing pharmaceutical composition to be used in tissue regeneration will be determined by the attending physician considering various factors which modify the action of the proteins, e.g., amount of tissue weight desired to be formed, the site of damage, the condition of the damaged tissue, the size of a wound, type of damaged tissue (e.g., bone), the patient 's age, sex, and diet, the severity of any infection, time of administration and other clinical factors.
  • the dosage may vary with the type of matrix used in the reconstitution and with inclusion of other proteins in the pharmaceutical composition. For example, the addition of other known growth factors, such as IGF I (insulin like growth factor I), to the final composition, may also effect the dosage.
  • IGF I insulin like growth factor I
  • 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 pu ⁇ oses. Patent and literature references cited herein are inco ⁇ orated by reference as if fully set forth.
  • Xaa Glu Met Lys Asp lie Ala He Asn He Ser Arg Asn Leu Lys Asp 65 70 75 80
  • AAATNTTAAA ATAATTCCAA GCTGAGTTTT CTAGATTGAG CAGAAATGGT GAAAGGAGTA 120
  • MOLECULE TYPE protein
  • GGACATGCCA GGAATAAAAA GGATACTCAC TGTTACCATT CTGGCTCTCT GTCTTCCAAG 240
  • CCCTGGGAAT GCACAGGCAC AGTGCACGAA TGGCTTTGAC CTGGATCGCC AGTCAGGACA 300 GTGTTTAGAT ATTGATGAAT GCCGAACCAT CCCCGAGGCC TGCCGAGGAG ACATGATGTG 360
  • NGACACTTAC TGGTTAAACT TACGTTGCTA AAGATTTCTC TATAATAAGC CACACATTAT 120
  • GAGTGTTCTT CAGAGGATCC TCGCTGCCCA GGTTCCCTGC CAGAAGGACA GAATAATCCT 240
  • GAGACTCCWC AATTATTGAT CCAGGAACTG AGCAAGATCT TCCTTCCCCT GAAAATAGTT 120
  • MOLECULE TYPE cDNA (xi) SEQUENCE DESCRIPTION: SEQ ID NO:28:
  • CCCCTCNTCC NTTTCCCCCC CAAGCACAGA GGGGAGAGGG GCCAGGGAAG TGGATGTTTC 60

Abstract

Novel proteins are disclosed.

Description

SECRETED PROTEINS
FIELD OF THE INVENTION
The present invention provides novel proteins, along with therapeutic, diagnostic and research utilities for these 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 cloning). More recent "indirect" cloning techniques such as signal sequence cloning, which isolates DNA sequences based on the presence of a now well-recognized secretory leader sequence motif, as well as various PCR-based or low stringency hybridization cloning techniques, have advanced the state of the art by making available large numbers of DNA/amino acid sequences for proteins that are known to have biological activity by virtue of their secreted nature in the case of leader sequence cloning, or by virtue of the cell or tissue source in the case of PCR-based techniques. It is to these proteins that the present invention is directed.
SUMMARY OF THE INVENTION In one embodiment, the present invention provides a composition comprising an isolated protein encoded by a polynucleotide selected from the group consisting of:
(a) a polynucleotide comprising the nucleotide sequence of SEQ ID NO: l;
(b) a polynucleotide comprising the nucleotide sequence of SEQ ID NO: 1 from nucleotide 70 to nucleotide 505;
(c) a polynucleotide comprising the nucleotide sequence of the full length protein coding sequence of clone API 62 deposited under accession number ATCC 98026 ;
(d) a polynucleotide encoding the full length protein encoded by the cDNA insert of clone AP162 deposited under accession number ATCC 98026 ; (e) a polynucleotide comprising the nucleotide sequence of the mature protein coding sequence of clone API 62 deposited under accession number ATCC 98026 ;
(0 a poiynucleotideencoding the mature protein encoded by the cDNA insert of clone AP162 deposited under accession number ATCC 98026 ;
(g) a polynucleotide encoding a protein comprising the amino acid sequence of SEQ ID NO:2;
(h) a polynucleotide encoding a protein comprising a fragment of the amino acid sequence of SEQ ID NO:2 having biological activity; (i) a polynucleotide which is an allelic variant of a polynucleotide of
(a)-(f) above; and
(j) a polynucleotide which encodes a species homologue of the protein of (g) or (h) above.
Preferably, such polynucleotide comprises the nucleotide sequence of SEQ ID NO: l from nucleotide 70 to nucleotide 505; the nucleotide sequence of the full length protein coding sequence of clone AP162 deposited under accession number ATCC 98026 ; or the nucleotide sequence of the mature protein coding sequence of clone API 62 deposited under accession number ATCC 98026 . In other preferred embodiments, the polynucleotide encodes the full length or mature protein encoded by the cDNA insert of clone AP162 deposited under accession number ATCC 98026 . In yet other preferred embodiments, such polynucleotide encodes a protein comprising the amino acid sequence of SEQ ID NO:2 from amino acid 42 to amino acid 61.
In other embodiments, the present invention provides a composition comprising a protein, wherein said protein comprises an amino acid sequence selected from the group consisting of:
(a) the amino acid sequence of SEQ ID NO:2;
(b) the amino acid sequence of SEQ ID NO:2 from amino acid 42 to amino acid 61 ;
(c) fragments of the amino a' 'd sequence of SEQ ID NO:2; and (d) the amino acid sequence encoded by the cDNA insert of clone
API 62 deposited under accession number ATCC 98026; the protein being substantially free from other mammalian proteins. Preferably such protein comprises the amino acid sequence of SEQ ID NO:2 or the amino acid sequence of SEQ ID NO:2 from amino acid 42 to amino acid 61. In one embodiment, the present invention provides a composition comprising an isolated protein encoded by a polynucleotide selected from the group consisting of:
(a) a polynucleotide comprising the nucleotide sequence of SEQ ID NO:4;
(b) a polynucleotide comprising the nucleotide sequence of SEQ ID NO:4 from nucleotide 230 to nucleotide 791 ;
(c) a polynucleotide comprising the nucleotide sequence of SEQ ID NO:4 from nucleotide 31 1 to nucleotide 791 ;
(d) a polynucleotide comprising the nucleotide sequence of the full length protein coding sequence of clone AM931 deposited under accession number ATCC 98026 ;
(e) a polynucleotide encoding the full length protein encoded by the cDNA insert of clone AM931 deposited under accession number ATCC 98026 ;
(f) a polynucleotide comprising the nucleotide sequence of the mature protein coding sequence of clone AM931 deposited under accession number ATCC 98026 ;
(g) a polynucleotideencoding the mature protein encoded by the cDNA insert of clone AM931 deposited under accession number ATCC 98026 ;
(h) a polynucleotide encoding a protein comprising the amino acid sequence of SEQ ID NO:5; (i) a polynucleotide encoding a protein comprising a fragment of the amino acid sequence of SEQ ID NO:5 having biological activity;
(j) a polynucleotide which is an allelic variant of a polynucleotide of (a)-(g) above; and
(k) a polynucleotide which encodes a species homologue of the protein of (h) or (i) above.
Preferably, such polynucleotide comprises the nucleotide sequence of SEQ ID NO:4 from nucleotide 230 to nucleotide 791; the nucleotide sequence of SEQ ID NO:4 from nucleotide 31 1 to nucleotide 791 ; the nucleotide sequence of the full length protein coding sequence of clone AM931 deposited under accession number ATCC 98026 ; or the nucleotide sequence of the mature protein coding sequence of clone AM931 deposited under accession number ATCC 98026 . In other preferred embodiments, the polynucleotide encodes the full length or mature protein encoded by the cDNA insert of clone AM931 deposited under accession number ATCC 98026 . In yet other preferred embodiments, such polynucleotide encodes a protein comprising the amino acid sequence of SEQ ID NO:5 from amino acid 32 to amino acid 51.
In other embodiments, the present invention provides a composition comprising a protein, wherein said protein comprises an amino acid sequence selected from the group consisting of: (a) the amino acid sequence of SEQ ID NO:5;
(b) the amino acid sequence of SEQ ID NO:5 from amino acid 32 to amino acid 51 ;
(c) fragments of the amino acid sequence of SEQ ID NO:5; and
(d) the amino acid sequence encoded by the cDNA insert of clone AM931 deposited under accession number ATCC 98026; the protein being substantially free from other mammalian proteins. Preferably such protein comprises the amino acid sequence of SEQ ID NO:5 or the amino acid sequence of SEQ ID NO:5 from amino acid 32 to amino acid 51.
In one embodiment, the present invention provides a composition comprising an isolated protein encoded by a polynucleotide selected from the group consisting of:
(a) a polynucleotide comprising the nucleotide sequence of SEQ ID NO:6;
(b) a polynucleotide comprising the nucleotide sequence of SEQ ID NO:6 from nucleotide 14 to nucleotide 491 ; (c) a polynucleotide comprising the nucleotide sequence of SEQ ID
NO:6 from nucleotide 83 to nucleotide 491 ;
(d) a polynucleotide comprising the nucleotide sequence of the full length protein coding sequence of clone AM610 deposited under accession number ATCC 98026 ; (e) a polynucleotide encoding the full length protein encoded by the cDNA insert of clone AM610 deposited under accession number ATCC 98026 ; (f) a polynucleotide comprising the nucleotide sequence of the mature protein coding sequence of clone AM610 deposited under accession number ATCC 98026 ; (g) a polynucleotideencoding the mature protein encoded by the cDNA insert of clone AM610 deposited under accession number ATCC 98026 ,
(h) a polynucleotide encoding a protein comprising the amino acid sequence of SEQ ID NO 7, (i) a polynucleotide encoding a protein compπsing a fragment of the ammo acid sequence of SEQ ID NO 7 having biological activity,
(j) a polynucleotide which is an allelic variant of a polynucleotide of
(a)-(g) above, and
(k) a polynucleotide which encodes a species homologue of the protein of (h) or (l) above
Preferably, such polynucleotide comprises the nucleotide sequence of SEQ ID NO 6 from nucleotide 14 to nucleotide 491 , the nucleotide sequence of SEQ ID NO 6 from nucleotide 83 to nucleotide 491 , the nucleotide sequence of the full length protein coding sequence of clone AM610 deposited under accession number ATCC 98026 , or the nucleotide sequence of the mature protein coding sequence of clone AM610 deposited under accession number ATCC 98026 In other preferred embodiments, the polynucleotide encodes the full length or mature protein encoded by the cDNA insert of clone AM610 deposited under accession number ATCC 98026 In yet other preferred embodiments, such polynucleotide encodes a protein comprising the amino acid sequence of SEQ ID NO 7 from ammo acid 31 to ammo acid 50
In other embodiments, the present invention provides a composition comprising a protein, wherein said protein comprises an ammo acid sequence selected from the group consisting of
(a) the amino acid sequence of SEQ ID NO 7, (b) the ammo acid sequence of SEQ ID NO 7 from ammo acid 31 to amino acid 50,
(c) fragments of the ammo acid sequence of SEQ ID NO 7, and
(d) the amino acid sequence encoded by the cDNA insert of clone AM610 deposited under accession number ATCC 98026, the protein being substantially free from other mammalian proteins Preferably such protein comprises the ammo acid sequence of SEQ ID NO 7 or the ammo acid sequence of SEQ ID NO 7 from ammo acid 31 to ammo acid 50
In one embodiment, the present invention provides a composition comprising an isolated protein encoded by a 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 1 to nucleotide 483, (c) a polynucleotide comprising the nucleotide sequence of the full length protein coding sequence of clone AM340 deposited under accession number ATCC 98026 ,
(d) a polynucleotide encoding the full length protein encoded by the cDNA insert of clone AM340 deposited under accession number ATCC 98026 , (e) a polynucleotide compπsing the nucleotide sequence of the mature protein coding sequence of clone AM340 deposited under accession number ATCC 98026 ,
(0 a polynucleotideencoding the mature protein encoded by the cDNA insert of clone AM340 deposited under accession number ATCC 98026 , (g) a polynucleotide encoding a protein comprising the ammo acid sequence of SEQ ID NO 10,
(h) a polynucleotide encoding a protein comprising a fragment of the ammo acid sequence of SEQ ID NO 10 having biological activity,
(0 a polynucleotide which is an allelic variant of a polynucleotide of (a)-(0 above, and
(j) a polynucleotide which encodes a species homologue of the protein of (g) or (h) above
Preferably, such polynucleotidecompπses the nucleotide sequence of SEQ ID NO 9 from nucleotide 1 to nucleotide 483, the nucleotide sequence of the full length protein coding sequence of clone AM340 deposited under accession number ATCC 98026 , or the nucleotide sequence of the mature protein coding sequence of clone AM340 deposited under accession number ATCC 98026 In other preferred embodiments, the polynucleotide encodes the full length or mature protein encoded by the cDNA insert of clone AM340 deposited under accession number ATCC 98026 In yet other preferred embodiments, such polynucleotide encodes a protein comprising the ammo acid sequence of SEQ ID
NO.10 from amino acid 124 to amino acid 143
In other embodiments, the present invention provides a composition comprising a protein, wherein said protein compπses an amino acid sequence selected from the group consisting of (a) the amino acid sequence of SEQ ID NO: 10,
(b) the ammo acid sequence of SEQ ID NO: 10 from ammo acid 124
Figure imgf000009_0001
(c) fragments of the amino acid sequence of SEQ ID NO: 10; and (d) the amino acid sequence encoded by the cDNA insert of clone
AM340 deposited under accession number ATCC 98026, (he protein being substantially free from other mammalian proteins. Preferably such protein comprises the amino acid sequence of SEQ ID NO- 10 or the amino acid sequence of SEQ ID NO.10 from ammo acid 124 to ammo acid 143 In one embodiment, the present invention provides a composition comprising an isolated protein encoded by a polynucleotide selected from the group consisting of
(a) a polynucleotide comprising the nucleotide sequence of SEQ ID NO. l l ,
(b) a polynucleotide comprising the nucleotide sequence of SEQ ID NO. l 1 from nucleotide 15 to nucleotide 462,
(c) a polynucleotide comprising the nucleotide sequence of SEQ ID NO: l 1 from nucleotide 87 to nucleotide 462,
(d) a polynucleotide comprising the nucleotide sequence of the full length protein coding sequence of clone AM282 deposited under accession number ATCC 98026 ,
(e) a polynucleotide encoding the full length protein encoded by the cDNA insert of clone AM282 deposited under accession number ATCC 98026 ,
(f) a polynucleotide comprising the nucleotide sequence of the mature protein coding sequence of clone AM282 deposited under accession number ATCC 98026 ,
(g) a polynucleotideencoding the mature protein encoded by the cDNA insert of clone AM282 deposited under accession number ATCC 98026 ,
(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;
(j) a polynucleotide which is an allelic vaπant of a polynucleotide of
(a)-(g) above; and (k) a polynucleotide which encodes a species homologue of the protein of (h) or (i) above
Preferably, such polynucleotide comprises the nucleotide sequence of SEQ ID NO 1 1 from nucleotide 15 to nucleotide 462, the nucleotide sequence of SEQ ID NO.11 from nucleotide 87 to nucleotide 462, the nucleotide sequence of the full length protein coding sequence of clone AM282 deposited under accession number ATCC 98026 , or the nucleotide sequence of the mature protein coding sequence of clone AM282 deposited under accession number ATCC 98026 In other preferred embodiments, the polynucleotide encodes the full length or mature protein encoded by the cDNA insert of clone AM282 deposited under accession number ATCC 98026 In yet other preferred embodiments, such polynucleotide encodes a protein comprising the ammo acid sequence of SEQ ID NO 12 from ammo acid 28 to amino acid 47
In other embodiments, the present invention provides a composition comprising a protein, wherem said protein comprises an amino acid sequence selected from the group consisting of
(a) the amino acid sequence of SEQ ID NO.12,
(b) the ammo acid sequence of SEQ ID NO 12 from amino acid 28 to
Figure imgf000010_0001
(c) fragments of the amino acid sequence of SEQ ID NO 12, and (d) the amino acid sequence encoded by the cDNA insert of clone
AM282 deposited under accession number ATCC 98026, the protein being substantially free from other mammalian proteins Preferably such protein comprises the ammo acid sequence of SEQ ID NO 12 or the ammo acid sequence of SEQ ID NO: 12 from ammo acid 28 to ammo acid 47 In one embodiment, the present invention provides a composition compπsing an isolated protein encoded by a polynucleotide selected from the group consisting of
(a) a polynucleotide compπsing the nucleotide sequence of SEQ ID NO: 14,
(b) a polynucleotide comprising the nucleotide sequence of SEQ ID NO 14 from nucleotide 185 to nucleotide 519,
(c) a polynucleotide comprising the nucleotide sequence of SEQ ID NO.14 from nucleotide 260 to nucleotide 519, (d) a polynucleotide comprising the nucleotide sequence of the full length protein coding sequence of clone AK647 deposited under accession number ATCC 98026 ,
(e) a polynucleotide encoding the full length protein encoded by the cDNA insert of clone AK647 deposited under accession number ATCC 98026 ,
(0 a polynucleotide comprising the nucleotide sequence of the mature protein coding sequence of clone AK647 deposited under accession number ATCC 98026 ,
(g) a polynucleotideencodmg the mature protein encoded by the cDNA insert of clone AK647 deposited under accession number ATCC 98026 ,
(h) a polynucleotide encoding a protein comprising the amino acid sequence of SEQ ID NO 15,
(0 a polynucleotide encoding a protem comprising a fragment of the amino acid sequence of SEQ ID NO 15 having biological activity, (j) a polynucleotide which is an allelic vaπant of a polynucleotide of
(a)-(g) above, and
(k) a polynucleotide which encodes a species homologue of the protem of (h) or (i) above
Preferably, such polynucleotide comprises the nucleotide sequence of SEQ ID NO 14 from nucleotide 185 to nucleotide 519, the nucleotide sequence of SEQ ID NO 14 from nucleotide 260 to nucleotide 519, the nucleotide sequence of the full length protein coding sequence of clone AK647 deposited under accession number ATCC 98026 , or the nucleotide sequence of the mature protein coding sequence of clone AK647 deposited under accession number ATCC 98026 In other preferred embodiments, the polynucleotide encodes the full length or mature protein encoded by the cDNA insert of clone AK647 deposited under accession number ATCC 98026 In yet other preferred embodiments, such polynucleotide encodes a protein comprising the ammo acid sequence of SEQ ID NO- 15 from amino acid 27 to ammo acid 46
In other embodiments, the present invention provides a composition comprising a protein, wherein said protein comprises an ammo acid sequence selected from the group consisting of
(a) the amino acid sequence of SEQ ID NO 15,
(b) the ammo acid sequence of SEQ ID NO 15 from ammo acid 27 to ammo acid 46, (c) fragments of the ammo acid sequence of SEQ ID NO 15, and
(d) the ammo acid sequence encoded by the cDNA insert of clone AK647 deposited under accession number ATCC 98026, the protein being substantially free from other mammalian proteins Preferably such protein comprises the ammo acid sequence of SEQ ID NO 15 or the amino acid sequence of SEQ ID NO 15 from amino acid 27 to ammo acid 46
In one embodiment, the present invention provides a composition compπsing an isolated protem encoded by a polynucleotide selected from the group consisting of
(a) a polynucleotide comprising the nucieotide sequence of SEQ ID NO 17,
(b) a polynucleotide comprising the nucleotide sequence of SEQ ID NO 17 from nucleotide 257 to nucleotide 536,
(c) a polynucleotide comprising the nucleotide sequence of SEQ ID NO 17 from nucleotide 329 to nucleotide 536, (d) d polynucleotide comprising the nucleotide sequence of the full length protein coding sequence of clone AK583 deposited under accession number ATCC 98026 ,
(e) a polynucleotide encoding the full length protein encoded by the cDNA insert of clone AK583 deposited under accession number ATCC 98026 , (f) a polynucleotide comprising the nucleotide sequence of the mature protein coding sequence of clone AK583 deposited under accession number ATCC 98026 ,
(g) a polynucleotideencoding the mature protem encoded by the cDNA insert of clone AK583 deposited under accession number ATCC 98026 , (h) a polynucleotide encoding a protein compπsing the ammo acid sequence of SEQ ID NO 18,
(l) a polynucleotide encoding a protem comprising a fragment of the amino acid sequence of SEQ ID NO 18 having biological activity,
(j) a polynucleotide which is an allelic variant of a polynucleotide of (a)-(g) above, and
(k) a polynucleotide which encodes a species homologue of the protein of (h) or (I) above
Preferably, such polynucleotide comprises the nucleotide sequence of SEQ ID NO 17 from nucleotide 257 to nucleotide 536, the nucleotide sequence of SEQ ID NO 17 from nucleotide 329 to nucleotide 536; the nucleotide sequence of the full length protein coding sequence of clone AK583 deposited under accession number ATCC 98026 ; or the nucleotide sequence of the mature protein coding sequence of clone AK583 deposited under accession number ATCC 98026 In other preferred embodiments, the polynucleotide encodes the full length or mature protein encoded by the cDNA insert of clone AK583 deposited under accession number ATCC 98026 In yet other preferred embodiments. Such polynucleotide encodes a protein compπsing the amino acid sequence of SEQ ID NO 18 from ammo acid 14 to amino acid 33
In other embodiments, the present invention provides a composition comprising a protein, wherein said protein comprises an ammo acid sequence selected from the group consisting of
(a) the amino acid sequence of SEQ ID NO.18,
(b) the ammo acid sequence of SEQ ID NO 18 from amino acid 14 to ammo acid 33, (c) fragments of the ammo acid sequence of SEQ ID NO 18; and
(d) the amino acid sequence encoded by the cDNA insert of clone AK583 deposited under accession number ATCC 98026; the protein being substantially free from other mammalian proteins Preferably such protein comprises the amino acid sequence of SEQ ID NO- 18 or the ammo acid sequence of SEQ ID NO 18 from amino acid 14 to amino acid 33
In one embodiment, the present invention provides a composition comprising an isolated protein encoded by a polynucleotide selected from the group consisting of
(a) a polynucleotide comprising the nucleotide sequence of SEQ ID NO-20, (b) a polynucleotide comprising the nucleotide sequence of SEQ ID
NO:20 from nucleotide 179 to nucleotide 476,
(c) a polynucleotide compπsing the nucleotide sequence of the full length protein coding sequence of clone AK533 deposited under accession number ATCC 98026 , (d) a polynucleotide encoding the full length protem encoded by the cDNA insert of clone AK533 deposited under accession number ATCC 98026 ;
(e) a polynucleotide comprising the nucleotide sequence of the mature protein coding sequence of clone AK533 deposited under accession number ATCC 98026 , (0 a polynucleotideencodingthe mature protein encoded by the cDNA insert of clone AK533 deposited under accession number ATCC 98026 ,
(g) a polynucleotide encoding a protein comprising the ammo acid sequence of SEQ ID NO 21 , (h) a polynucleotide encoding a protein comprising a fragment of the amino acid sequence of SEQ ID NO 21 having biological activity,
(0 a polynucleotide which is an allelic variant of a polynucleotide of
(a)-(f) above, and
(j) a polynucleotide which encodes a species homologue of the protein of (g) or (h) above
Preferably, such polynucleotide comprises the nucleotide sequence of SEQ ID NO 20 from nucleotide 179 to nucleotide 476, the nucleotide sequence of the full length protein coding sequence of clone AK533 deposited under accession number ATCC 98026 , or the nucleotide sequence of the mature protein coding sequence of clone AK533 deposited under accession number ATCC 98026 In other preferred embodiments, the polynucleotide encodes the full length or mature protein encoded by the cDNA insert of clone AK533 deposited under accession number ATCC 98026 In yet other preferred embodiments, such polynucleotide encodes a protein compπsing the amino acid sequence of SEQ ID NO 21 from amino acid 35 to amino acid 57 In other embodiments, the present invention provides a composition comprising a protein, wherein said protein comprises an ammo acid sequence selected from the group consisting of
(a) the amino acid sequence of SEQ ID NO 21 ,
(b) the ammo acid sequence of SEQ ID NO 21 from ammo acid 35 to
Figure imgf000014_0001
(c) fragments of the amino acid sequence of SEQ ID NO 21 , and
(d) the amino acid sequence encoded by the cDNA insert of clone AK533 deposited under accession number ATCC 98026, the protein being substantially free from other mammalian proteins Preferably such protein comprises the amino acid sequence of SEQ ID NO 21 or the ammo acid sequence of SEQ
ID NO 21 from amino acid 35 to ammo acid 57
In one embodiment, the present invention provides a composition comprising an isolated protein encoded by a polynucleotide selected from the group consisting of (a) a polynucleotide comprising the nucleotide sequence of SEQ ID NO:23;
(b) a polynucleotide comprising the nucleotide sequence of SEQ ID NO:23 from nucleotide 220 to nucleotide 612; (c) a polynucleotide comprising the nucleotide sequence of SEQ ID
NO:23 from nucleotide 328 to nucleotide 612;
(d) a polynucleotide comprising the nucleotide sequence of the full length protein coding sequence of clone AK296 deposited under accession number ATCC 98026 ; (e) a polynucleotide encoding the full length protein encoded by the cDNA insert of clone AK296 deposited under accession number ATCC 98026 ;
(0 a polynucleotide comprising the nucleotide sequence of the mature protein coding sequence of clone AK296 deposited under accession number ATCC 98026 ; (g) a polynucleotideencoding the mature protein encoded by the cDNA insert of clone AK296 deposited under accession number ATCC 98026 ;
(h) a polynucleotide encoding a protein comprising the amino acid sequence of SEQ ID NO:24;
(i) a polynucleotide encoding a protein comprising a fragment of the amino acid sequence of SEQ ID NO:24 having biological activity;
(j) a polynucleotide which is an allelic variant of a polynucleotide of
(a)-(g) above; and
(k) a polynucleotide which encodes a species homologue of the protein of (h) or (i) above. Preferably, such polynucleotide comprises the nucleotide sequence of SEQ ID
NO:23 from nucleotide 220 to nucleotide 612; the nucleotide sequence of SEQ ID NO:23 from nucleotide 328 to nucleotide 612; the nucleotide sequence of the full length protein coding sequence of clone AK296 deposited under accession number ATCC 98026 ; or the nucleotide sequence of the mature protein coding sequence of clone AK296 deposited under accession number ATCC 98026 . In other preferred embodiments, the polynucleotide encodes the full length or mature protein encoded by the cDNA insert of clone AK296 deposited under accession number ATCC 98026 . In yet other preferred embodiments, such polynucleotide encodes a protein comprising the amino acid sequence of SEQ ID NO:24 from amino acid 81 to amino acid 90. In other embodiments, the present invention provides a composition comprising a protein, wherein said protein comprises an amino acid sequence selected from the group consisting of:
(a) the amino acid sequence of SEQ ID NO:24; (b) the amino acid sequence of SEQ ID NO:24 from amino acid 81 to amino acid 90;
(c) fragments of the amino acid sequence of SEQ ID NO:24; and
(d) the amino acid sequence encoded by the cDNA insert of clone AK296 deposited under accession number ATCC 98026; the protein being substantially free from other mammalian proteins. Preferably such protein comprises the amino acid sequence of SEQ ID NO:24 or the amino acid sequence of SEQ ID NO:24 from amino acid 81 to amino acid 90.
In one embodiment, the present invention provides a composition comprising an isolated protein encoded by a polynucleotide selected from the group consisting of: (a) a polynucleotide comprising the nucleotide sequence of SEQ ID
NO:26;
(b) a polynucleotide comprising the nucleotide sequence of SEQ ID NO:26 from nucleotide 58 to nucleotide 655;
(c) a polynucleotide comprising the nucleotide sequence of the full length protein coding sequence of clone H617 deposited under accession number
ATCC 98026 ;
(d) a polynucleotide encoding the full length protein encoded by the cDNA insert of clone H617 deposited under accession number ATCC 98026 ;
(e) a polynucleotide comprising the nucleotide sequence of the mature protein coding sequence of clone H617 deposited under accession number ATCC
98026 ;
(f) a polynucleotideencoding the mature protein encoded by the cDNA insert of clone H617 deposited under accession number ATCC 98026 ;
(g) a polynucleotide encoding a protein comprising the amino acid sequence of SEQ ID NO:27;
(h) a polynucleotide encoding a protein comprising a fragment of the amino acid sequence of SEQ ID NO:27 having biological activity;
(i) a polynucleotide which is an allelic variant of a polynucleotide of
(a)-(f) above; and (j) a polynucleotide which encodes a species homologue of the protein of (g) or (h) above
Preferably, such polynucleotide comprises the nucleotide sequence of SEQ ID NO.26 from nucleotide 58 to nucleotide 655. the nucleotide sequence of the full length protein coding sequence of clone H617 deposited under accession number ATCC 98026 ; or the nucleotide sequence of the mature protein coding sequence of clone H617 deposited uVider accession number ATCC 98026 In other preferred embodiments, the polynucleotide encodes the full length or mature protein encoded by the cDNA insert of clone H617 deposited under accession number ATCC 98026 In yet other preferred embodiments, such polynucleotide encodes a protein comprising the amino acid sequence of SEQ ID NO 27 from amino acid 65 to amino acid 84
In other embodiments, the present invention provides a composition compπsing a protein, wherein said protein compπses an amino acid sequence selected from the group consisting of (a) the amino acid sequence of SEQ ID NO.27,
(b) the amino acid sequence of SEQ ID NO 27 from ammo acid 65 to
Figure imgf000017_0001
(c) fragments of the amino acid sequence of SEQ ID NO 27; and
(d) the amino acid sequence encoded by the cDNA insert of clone H617 deposited under accession number ATCC 98026, the protein being substantially free from other mammalian proteins Preferably such protein comprises the amino acid sequence of SEQ ID NO 27 or the amino acid sequence of SEQ ID NO 27 from amino acid 65 to amino acid 84
In one embodiment, the present invention provides a composition comprising an isolated protein encoded by a polynucleotide selected from the group consisting of
(a) a polynucleotide comprising the nucleotide sequence of SEQ ID NO:29,
(b) a polynucleotide comprising the nucleotide sequence of SEQ ID NO 29 from nucleotide 14 to nucleotide 391 , (c) a polynucleotide compπsing the nucleotide sequence of the full length protein coding sequence of clone BB9 deposited under accession number ATCC 98026 ,
(d) a polynucleotide encoding the full length protein encoded by the cDNA insert of clone BB9 deposited under accession number ATCC 98026 , (e) a polynucleotide comprising the nucleotide sequence of the mature protein coding sequence of clone BB9 deposited under accession number ATCC 98026 ,
(f) a polynucleotideencoding the mature protein encoded by the cDNA insert of clone BB9 deposited under accession number ATCC 98026 ,
(g) a polynucleotide encoding a protein comprising the ammo acid sequence of SEQ ID NO 30,
(h) a polynucleotide encoding a protein comprising a fragment of the amino acid sequence of SEQ ID NO 30 having biological activity, (i) a polynucleotide which is an allelic variant of a polynucleotide of
(a)-(0 above, and
(j) a polynucleotide which encodes a species homologue of the protein of (g) or (h) above
Preferably, such polynucleotide compπses the nucleotide sequence of SEQ ID NO 29 from nucleotide 14 to nucleotide 391 , the nucleotide sequence of the full length protein coding sequence of clone BB9 deposited under accession number ATCC 98026 , or the nucleotide sequence of the mature protein coding sequence of clone BB9 deposited under accession number ATCC 98026 In other preferred embodiments, the polynucleotide encodes the full length or mature protein encoded by the cDNA insert of clone BB9 deposited under accession number ATCC 98026 In yet other preferred embodiments, such polynucleotide encodes a protein comprising the amino acid sequence of SEQ ID NO 30 from amino acid 75 to ammo acid 94
In other embodiments, the present invention provides a composition comprising a protein, wherein said protein comprises an amino acid sequence selected from the group consisting of
(a) the amino acid sequence of SEQ ID NO.30,
(b) the amino acid sequence of SEQ ID NO 30 from amino acid 75 to amino acid 94;
(c) fragments of the amino acid sequence of SEQ ID NO 30, and (d) the amino acid sequence encoded by the cDN A insert of clone BB9 deposited under accession number ATCC 98026, the protein being substantially free from other mammalian proteins. Preferably such protein comprises the amino acid sequence of SEQ ID NO 30 or the amino acid sequence of SEQ ID NO.30 from amino acid 75 to ammo acid 94 In one embodiment, the present invention provides a composition comprising an isolated protein encoded by a polynucleotide selected from the group consistmg of
(a) a polynucleotide comprising the nucleotide sequence of SEQ ID NO 32, (b) a polynucleotide comprising the nucleotide sequence of SEQ ID
NO 32 from nucleotide 61 to nucleotide 514,
(c) a polynucleotide comprising the nucleotide sequence of SEQ ID NO 32 from nucleotide 115 to nucleotide 514,
(d) a polynucleotide comprising the nucleotide sequence of the full length protein coding sequence of clone AW 191 deposited under accession number
ATCC 98026 ,
(e) a polynucleotide encoding the full length protein encoded by the cDNA insert of clone AW191 deposited under accession number ATCC 98026 ,
(f) a polynucleotide comprising the nucleotide sequence of the mature protein coding sequence of clone AW 191 deposited under accession number ATCC
98026 ,
(g) a polynucleotideencoding the mature protein encoded by the cDNA insert of clone AW191 deposited under accession number ATCC 98026 ,
(h) a polynucleotide encoding a protein comprising the ammo acid sequence of SEQ ID NO 33,
(0 a polynucleotide encoding a protein compπsing a fragment of the ammo acid sequence of SEQ ID NO 33 having biological activity,
0) a polynucleotide which is an allelic variant of a polynucleotide of
(a)-(g) above, and (k) a polynucleotide which encodes a species homologue of the protein of (h) or (0 above
Preferably, such polynucleotide comprises the nucleotide sequence of SEQ ID NO 32 from nucleotide 61 to nucleotide 514, the nucleotide sequence of SEQ ID NO 32 from nucleotide 1 15 to nucleotide 514, the nucleotide sequence of the full length protein coding sequence of clone AW191 deposited under accession number ATCC 98026 , or the nucleotide sequence of the mature protein coding sequence of clone AW 191 deposited under accession number ATCC 98026 In other preferred embodiments, the polynucleotide encodes the full length or mature protein encoded by the cDNA insert of clone AW191 deposited under accession number ATCC 98026 In yet other preferred embodiments, such polynucleotide encodes a protein comprising the amino acid sequence of SEQ ID NO:33 from amino acid 24 to amino acid 43.
In other embodiments, the present invention provides a composition comprising a protein, wherein said protein comprises an amino acid sequence selected from the group consisting of:
(a) the amino acid sequence of SEQ ID NO:33;
(b) the amino acid sequence of SEQ ID NO:33 from amino acid 24 to amino acid 43;
(c) fragments of the amino acid sequence of SEQ ID NO:33; and (d) the amino acid sequence encoded by the cDNA insert of clone
AW191 deposited under accession number ATCC 98026; the protein being substantially free from other mammalian proteins. Preferably such protein comprises the amino acid sequence of SEQ ID NO:33 or the amino acid sequence of SEQ
ID NO:33 from amino acid 24 to amino acid 43. In one embodiment, the present invention provides a composition comprising an isolated protein encoded by a polynucleotide selected from the group consisting of:
(a) a polynucleotide comprising the nucleotide sequence of SEQ ID NO:35;
(b) a polynucleotide comprising the nucleotide sequence of SEQ ID NO:35 from nucleotide 180 to nucleotide 525;
(c) a polynucleotide comprising the nucleotide sequence of SEQ ID NO:35 from nucleotide 339 to nucleotide 525;
(d) a polynucleotide comprising the nucleotide sequence of the full length protein coding sequence of clone ΛT21 1 deposited under accession number ATCC 98026 ;
(e) a polynucleotide encoding the full length protein encoded by the cDNA insert of clone AT211 deposited under accession number ATCC 98026 ;
(f) a polynucleotide comprising the nucleotide sequence of the mature protein coding sequence of clone AT211 deposited under accession number ATCC 98026 ;
(g) a polynucleotideencoding the mature protein encoded by the cDNA insert of clone AT21 1 deposited under accession number ATCC 98026 ;
(h) a polynucleotide encoding a protein comprising the amino acid sequence of SEQ ID NO:36; (1) a polynucleotide encoding a protein compπsing a fragment of the amino acid sequence of SEQ ID NO 36 having biological activity,
(j) a polynucleotide which is an allelic variant of a polynucleotide of
(a)-(g) above, and (k) a polynucleotide which encodes a species homologue of the protein of (h) or (l) above
Preferably, such polynucleotide compπses the nucleotide sequence of SEQ ID NO 35 from nucleotide 180 to nucleotide 525, the nucleotide sequence of SEQ ID NO 35 from nucleotide 339 to nucleotide 525, the nucleotide sequence of the full length protein coding sequence of clone AT211 deposited under accession number ATCC 98026 , or the nucleotide sequence of the mature protein coding sequence of clone AT211 deposited under accession number ATCC 98026 In other preferred embodiments, the polynucleotide encodes the full length or mature protein encoded by the cDNA insert of clone AT211 deposited under accession number ATCC 98026 In yet other preferred embodiments, such polynucleotide encodes a protein comprising the amino acid sequence of SEQ ID NO 36 from amino acid 1 to amino acid 20
In other embodiments, the present invention provides a composition compπsing a protein, wherein said protein comprises an ammo acid sequence selected from the group consisting of (a) the amino acid sequence of SEQ ID NO 36,
(b) the amino acid sequence of SEQ ID NO 36 from amino acid 1 to amino acid 20,
(c) fragments of the amino acid sequence of SEQ ID NO 36, and
(d) the ammo acid sequence encoded by the cDNA insert of clone AT211 deposited under accession number ATCC 98026. the protein being substantially free from other mammalian proteins Preferably such protein comprises the ammo acid sequence of SEQ ID NO 36 or the ammo acid sequence of SEQ ID NO 36 from amino acid 1 to amino acid 20
In one embodiment, the present invention provides a composition comprising an isolated protein encoded by a polynucleotide selected from the group consisting of
(a) a polynucleotide comprising the nucleotide sequence of SEQ ID NO 38,
(b) a polynucleotide comprising the nucleotide sequence of SEQ ID NO 38 from nucleotide 225 to nucleotide 677, (c) a polynucleotide comprising the nucleotide sequence of SEQ ID NO:38 from nucleotide 390 to nucleotide 677;
(d) a polynucleotide comprising the nucleotide sequence of the full length protein coding sequence of clone AT205 deposited under accession number ATCC 98026 ;
(e) a polynucleotide encoding the full length protein encoded by the cDNA insert of clone AT205 deposited under accession number ATCC 98026 ;
(0 a polynucleotide comprising the nucleotide sequence of the mature protein coding sequence of clone AT205 deposited under accession number ATCC 98026 ;
(g) a polynucleotideencoding the mature protein encoded by the cDNA insert of clone AT205 deposited under accession number ATCC 98026 ;
(h) a polynucleotide encoding a protein comprising the ammo acid sequence of SEQ ID NO: 39; (i) a polynucleotide encoding a protein comprising a fragment of the amino acid sequence of SEQ ID NO:39 having biological activity;
(j) a polynucleotide which is an allelic variant of a polynucleotide of
(a)-(g) above; and
(k) a polynucleotide which encodes a species homologue of the protein of (h) or (i) above.
Preferably, such polynucleotide comprises the nucleotide sequence of SEQ ID NO:38 from nucleotide 225 to nucleotide 677; the nucleotide sequence of SEQ ID NO:38 from nucleotide 390 to nucleotide 677; the nucleotide sequence of the full length protein coding sequence of clone AT205 deposited under accession number ATCC 98026 ; or the nucleotide sequence of the mature protein coding sequence of clone AT205 deposited under accession number ATCC 98026 . In other preferred embodiments, the polynucleotide encodes the full length or mature protein encoded by the cDNA insert of clone AT205 deposited under accession number ATCC 98026 . In yet other preferred embodiments, such polynucleotide encodes a protein comprising the amino acid sequence of SEQ ID NO:39 from amino acid 6 to amino acid 25.
In other embodiments, the present invention provides a composition comprising a protein, wherein said protein comprises an amino acid sequence selected from the group consisting of:
(a) the amino acid sequence of SEQ ID NO:39; (b) the ammo acid sequence of SEQ ID NO 39 from ammo acid 6 to ammo acid 25,
(c) fragments of the ammo acid sequence of SEQ ID NO 39, and
(d) the amino acid sequence encoded by the cDNA insert of clone AT205 deposited under accession number ATCC 98026, the protem being substantially free from other mammalian proteins Preferably such protein όompπses the ammo acid sequence of SEQ ID NO 39 or the amino acid sequence of SEQ ID NO 39 from ammo acid 6 to amino acid 25
In one embodiment, the present invention provides a composition comprising an isolated protein encoded by a polynucleotide selected from the group consisting of
(a) a polynucleotide comprising the nucleotide sequence of SEQ ID NO 40,
(b) a polynucleotide comprising the nucleotide sequence of SEQ ID NO 40 from nucleotide 128 to nucleotide 508, (c) a polynucleotide comprising the nucleotide sequence of SEQ ID
NO 40 from nucleotide 200 to nucleotide 508,
(d) a polynucleotide comprising the nucleotide sequence of the full length protein coding sequence of clone AS34 deposited under accession number ATCC 98026 , (e) a polynucleotide encoding the full length protein encoded by the cDNA insert of clone AS34 deposited under accession number ATCC 98026 ,
(0 a polynucleotide comprising the nucleotide sequence of the mature protein coding sequence of clone AS34 deposited under accession number ATCC 98026 , (g) a polynucleotideencoding the mature protein encoded by the cDNA insert of clone AS34 deposited under accession number ATCC 98026 ,
(h) a polynucleotide encoding a protein comprising the amino acid sequence of SEQ ID NO 41 ,
(0 a polynucleotide encoding a protein comprising a fragment of the ammo acid sequence of SEQ ID NO 41 having biological activity,
(j) a polynucleotide which is an allelic variant of a polynucleotide of
(a)-(g) above, and
(k) a polynucleotide which encodes a species homologue of the protein of (h) or (l) above Preferably, such polynucleotide comprises the nucleotide sequence of SEQ ID NO:40 from nucleotide 128 to nucleotide 508; the nucleotide sequence of SEQ ID NO:40 from nucleotide 200 to nucleotide 508; the nucleotide sequence of the full length protein coding sequence of clone AS34 deposited under accession number ATCC 98026 ; or the nucleotide sequence of the mature protein coding sequence of clone AS34 deposited under accession number ATCC 98026 . In other preferred embodiments, the polynucleotide eVicodes the full length or mature protein encoded by the cDNA insert of clone AS34 deposited under accession number ATCC 98026 . In yet other preferred embodiments, such polynucleotide encodes a protein comprising the amino acid sequence of SEQ ID NO:41 from amino acid 27 to amino acid 46.
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 ammo acid sequence of SEQ ID NO:41 ; (b) the amino acid sequence of SEQ ID NO:41 from amino acid 27 to amino acid 46;
(c) fragments of the amino acid sequence of SEQ ID NO:41 ; and
(d) the amino acid sequence encoded by the cDNA insert of clone AS34 deposited under accession number ATCC 98026; the protein being substantially free from other mammalian proteins. Preferably such protein comprises the amino acid sequence of SEQ ID NO:41 or the amino acid sequence of SEQ ID NO:41 from amino acid 27 to amino acid 46.
In one embodiment, the present invention provides a composition comprising an isolated protein encoded by a polynucleotide selected from the group consisting of: (a) a polynucleotide comprising the nucleotide sequence of SEQ ID
NO:43;
(b) a polynucleotide comprising the nucleotide sequence of SEQ ID NO:43 from nucleotide 23 to nucleotide 676;
(c) a polynucleotide comprising the nucleotide sequence of the full length p ;in coding sequence of clone AS32 deposited under accession number
ATCC 98026 ;
(d) a polynucleotide encoding the full length protein encoded by the cDNA insert of clone AS32 deposited under accession number ATCC 98026 ; (e) a polynucleotide comprising the nucleotide sequence of the mature protein coding sequence of clone AS32 deposited under accession number ATCC 98026 ;
(0 a polynucleotideencoding the mature protein encoded by the cDNA insert of clone AS32 deposited under accession number ATCC 98026 ;
(g) a polynucleotide encoding a protein comprising the amino acid sequence of SEQ ID NO:44;
(h) a polynucleotide encoding a protein comprising a fragment of the amino acid sequence of SEQ ID NO:44 having biological activity; (i) a polynucleotide which is an allelic variant of a polynucleotide of
(a)-(f) above; and
(j) a polynucleotide which encodes a species homologue of the protein of (g) or (h) above.
Preferably, such polynucleotide comprises the nucleotide sequence of SEQ ID NO:43 from nucleotide 23 to nucleotide 676; the nucleotide sequence of the full length protein coding sequence of clone AS32 deposited under accession number ATCC 98026 ; or the nucleotide sequence of the mature protein coding sequence of clone AS32 deposited under accession number ATCC 98026 . In other preferred embodiments, the polynucleotide encodes the full length or mature protein encoded by the cDNA insert of clone AS32 deposited under accession number ATCC 98026 . In yet other preferred embodiments, such polynucleotide encodes a protein comprising the amino acid sequence of SEQ ID NO:44 from amino acid 78 to amino acid 97.
In other embodiments, the present invention provides a composition comprising a protein, wherein said protein comprises an amino acid sequence selected from the group consisting of:
(a) the amino acid sequence of SEQ ID NO:44;
(b) the amino acid sequence of SEQ ID NO:44 from amino acid 78 to amino acid 97;
(c) fragments of the amino acid sequence of SEQ ID NO:44; and (d) the amino acid sequence encoded by the cDNA insert of clone
AS32 deposited under accession number ATCC 98026; the protein being substantially free from other mammalian proteins. Preferably such protein comprises the amino acid sequence of SEQ ID NO:44 or the amino acid sequence of SEQ ID NO:44 from amino acid 78 to amino acid 97. In one embodiment, the present invention provides a composition comprising an isolated protein encoded by a polynucleotide selected from the group consisting of
(a) a polynucleotide comprising the nucleotide sequence of SEQ ID NO 46, (b) a polynucleotide comprising the nucleotide sequence of SEQ ID
NO 46 from nucleotide 132 to nucleotide 479,
(c) a polynucleotide comprising the nucleotide sequence of SEQ ID NO 46 from nucleotide 201 to nucleotide 479,
(d) a polynucleotide comprising the nucleotide sequence of the full length protein coding sequence of clone AR260 deposited under accession number
ATCC 98026 ,
(e) a polynucleotide encoding the full length protein encoded by the cDNA insert of clone AR260 deposited under accession number ATCC 98026 ,
(0 a polynucleotide comprising the nucieotide sequence of the mature protein coding sequence of clone AR260 deposited under accession number ATCC
98026 ,
(g) a polynucleotide encoding the mature protein encoded by the cDNA insert of clone AR260 deposited under accession number ATCC 98026 ,
(h) a polynucleotide encoding a protein comprising the amino acid sequence of SEQ ID NO 47,
(i) a polynucleotide encoding a protein comprising a fragment of the ammo acid sequence of SEQ ID NO 47 having biological activity,
(j) a polynucleotide which is an allelic variant of a polynucleotide of
(a)-(g) above, and (k) a polynucleotide which encodes a species homologue of the protein of (h) or (I) above
Preferably, such polynucleotide comprises the nucleotide sequence of SEQ ID NO 46 from nucleotide 132 to nucleotide 479, the nucleotide sequence of SEQ ID NO 46 from nucleotide 201 to nucleotide 479, the nucleotide sequence of the full length protein coding sequence of clone AR260 deposited under accession number ATCC 98026 , or the nucleotide sequence of the mature protein coding sequence of clone AR260 deposited under accession number ATCC 98026 In other preferred embodiments, the polynucleotide encodes the full length or mature protein encoded by the cDNA insert of clone AR260 deposited under accession number ATCC 98026 In yet other preferred embodiments, such polynucleotide encodes a protein comprising the amino acid sequence of SEQ ID NO 47 from ammo acid 40 to amino acid 59
In other embodiments, the present invention provides a composition compπsing a protein, wherem said protein comprises an amino acid sequence selected from the group consisting of
(a) the ammo acid sequence of SEQ ID NO 47,
(b) the amino acid sequence of SEQ ID NO 47 from ammo acid 40 to amino acid 59,
(c) fragments of the amino acid sequence of SEQ ID NO 47, and (d) the ammo acid sequence encoded by the cDNA insert of clone
AR260 deposited under accession number ATCC 98026, the protein being substantially free from other mammalian proteins Preferably such protem comprises the amino acid sequence of SEQ ID NO 47 or the amino acid sequence of SEQ
ID NO 47 from amino acid 40 to amino acid 59 In one embodiment, the present invention provides a composition comprising an isolated protein encoded by a polynucleotide selected from the group consisting of
(a) a polynucleotide comprising the nucleotide sequence of SEQ ID NO 50,
(b) a polynucleotide comprising the nucleotide sequence of SEQ ID NO.50 from nucleotide 1 to nucleotide 332,
(c) a polynucleotide comprising the nucleotide sequence of the full length protein coding sequence of clone K640 deposited under accession number ATCC 98026 ,
(d) a polynucleotide encoding the full length protein encoded by the cDNA insert of clone K640 deposited under accession number ATCC 98026 ,
(e) a polynucleotide comprising the nucleotide sequence of the mature protein coding sequence of clone K640 deposited under accession number ATCC 98026 ,
(0 a polynucleotideencoding the mature protein encoded by the cDNA insert of clone K640 deposited under accession number ATCC 98026 ,
(g) a polynucleotide encoding a protein comprising the amino acid sequence of SEQ ID NO Sl ;
(h) a polynucleotide encoding a protein comprising a fragment of the ammo acid sequence of SEQ ID NO 51 having biological activity, (i) a polynucleotide which is an allelic variant of a polynucleotide of
(a)-(f) above, and
(j) a polynucleotide which encodes a species homologue of the protem of (g) or (h) above Preferably, such polynucleotide comprises the nucleotide sequence of SEQ ID
NO 50 from nucleotide 1 to nucleotide 332, the nucleotide sequence of the full length protein coding sequence of clone K640 deposited under accession number ATCC 98026 , or the nucleotide sequence of the mature protein coding sequence of clone K640 deposited under accession number ATCC 98026 In other preferred embodiments, the polynucleotide encodes the full length or mature protein encoded by the cDNA insert of clone K640 deposited under accession number ATCC 98026 In yet other preferred embodiments, such polynucleotide encodes a protein comprising the amino acid sequence of SEQ ID NO 51 from ammo acid 1 1 to amino acid 30
In other embodiments, the present invention provides a composition comprising a protein, wherein said protein comprises an amino acid sequence selected from the group consisting of
(a) the amino acid sequence of SEQ ID NO'51 ,
(b) the ammo acid sequence of SEQ ID NO 51 from ammo acid 1 1 to amino acid 30, (c) fragments of the amino acid sequence of SEQ ID NO 51 , and
(d) the ammo acid sequence encoded by the cDNA insert of clone K640 deposited under accession number ATCC 98026, the protem being substantially free from other mammalian proteins Preferably such protem comprises the amino acid sequence of SEQ ID NO 51 or the amino acid sequence of SEQ ID NO 51 from ammo acid 1 1 to ammo acid 30
In one embodiment, the present invention provides a composition comprising an isolated protein encoded by a polynucleotide selected from the group consisting of
(a) a polynucleotide comprising the nucleotide sequence of SEQ ID NO: 54, (b) a polynucleotide comprising the nucleotide sequence of SEQ' ID
NO- 54 from nucleotide 71 to nucleotide 377;
(c) a polynucleotide comprising the nucleotide sequence of the full length protein coding sequence of clone K39 deposited under accession number ATCC 98026 , (d) a polynucleotide encoding the full length protein encoded by the cDNA insert of clone K39 deposited under accession number ATCC 98026 ,
(e) a polynucleotide comprising the nucleotide sequence of the mature protein coding sequence of clone K39 deposited under accession number ATCC 98026 .
(0 a polynucleotideencoding the mature protein encoded by the cDNA insert of clone K39 deposited under accession number ATCC 98026 ,
(g) a polynucleotide encoding a protein comprising the ammo acid sequence of SEQ ID NO 55, (h) a polynucleotide encoding a protein comprising a fragment of the ammo acid sequence of SEQ ID NO 55 having biological activity,
(0 a polynucleotide which is an allelic variant of a polynucleotide of
(a)-(f) above, and
0) a polynucleotide which encodes a species homologue of the protem of (g) or (h) above
Preferably, such polynucleotide comprises the nucleotide sequence of SEQ ID NO 54 from nucleotide 71 to nucleotide 377, the nucleotide sequence of the full length protein coding sequence of clone K39 deposited under accession number ATCC 98026 , or the nucleotide sequence of the mature protein coding sequence of clone K39 deposited under accession number ATCC 98026 In other preferred embodiments, the polynucleotide encodes the full length or mature protein encoded by the cDNA insert of clone K39 deposited under accession number ATCC 98026 In yet other preferred embodiments, such polynucleotide encodes a protein comprising the amino acid sequence of SEQ ID NO 55 from amino acid 62 to amino acid 81 In other embodiments, the present invention provides a composition comprising a protein, wherein said protein comprises an amino acid sequence selected from the group consisting of
(a) the amino acid sequence of SEQ ID NO 55,
(b) the amino acid sequence of SEQ ID NO 55 from amino acid 62 to
Figure imgf000029_0001
,
(c) fragments of the ammo acid sequence of SEQ ID NO 55, and
(d) the ammo acid sequence encoded by the cDNA insert of clone K39 deposited under accession number ATCC 98026, the protein being substantially free from other mammalian proteins Preferably such protem comprises the ammo acid sequence of SEQ ID NO 55 or the amino acid sequence of SEQ ID NO 55 from ammo acid 62 to ammo acid 81
In one embodiment, the present invention provides a composition comprising an isolated protein encoded by a polynucleotide selected from the group consisting of
(a) a polynucleotide comprising the nucleotide sequence of SEQ ID NO 57,
(b) a polynucleotide comprising the nucleotide sequence of SEQ ID NO 57 from nucleotide 194 to nucleotide 423, (c) a polynucleotide comprising the nucleotide sequence of the full length protein coding sequence of clone AT319 deposited under accession number ATCC 98026 ,
(d) a polynucleotide encoding the full length protein encoded by the cDNA insert of clone AT319 deposited under accession number ATCC 98026 , (e) a polynucleotide compπsing the nucleotide sequence of the mature protein coding sequence of clone AT319 deposited under accession number ATCC 98026 ,
(f) a polynucleotideencoding the mature protein encoded by the cDNA insert of clone AT319 deposited under accession number ATCC 98026 , (g) a polynucleotide encoding a protem comprising the amino acid sequence of SEQ ID NO 58,
(h) a polynucleotide encoding a protem comprising a fragment of the ammo acid sequence of SEQ ID NO 58 having biological activity,
(0 a polynucleotide which is an allelic variant of a polynucleotide of (a)-(f) above, and
(j) a polynucleotide which encodes a species homologue of the protem of (g) or (h) above
Preferably, such polynucleotide compπses the nucleotide sequence of SEQ ID NO 57 from nucleotide 194 to nucleotide 423, the nucleotide sequence of the full length protein coding sequence of clone AT319 deposit. ' under accession number ATCC 98026
, or the nucleotide sequence of the mature protein coding sequence of clone AT319 deposited under accession number ATCC 98026 In other preferred embodiments, the polynucleotide encodes the full length or mature protein encoded by the cDNA insert of clone AT319 deposited under accession number ATCC 98026 In yet other preferred embodiments, such polynucleotide encodes a protem comprising the amino acid sequence of SEQ ID NO 58 from amino acid 2 to ammo acid 21
In other embodiments, the present mvention provides a composition comprising a protein, wherem said protein comprises an amino acid sequence selected from the group consisting of
(a) the ammo acid sequence of SEQ ID NO 58,
(b) the amino acid sequence of SEQ ID NO 58 from amino acid 2 to amino acid 21,
(c) fragments of the ammo acid sequence of SEQ ID NO 58, and (d) the ammo acid sequence encoded by the cDNA insert of clone
AT319 deposited under accession number ATCC 98026, the protein being substantially free from other mammalian proteins Preferably such protem comprises the ammo acid sequence of SEQ ID NO 58 or the ammo acid sequence of SEQ ID NO 58 from ammo acid 2 to ammo acid 21
Protein compositions of the present invention may further comprise a pharmaceutically acceptable carrier Compositions comprising an antibody which specifically reacts with such protein are also provided by the present invention
Methods are also provided for preventing, treating or ameliorating a medical condition which comprises administering to a mammalian subject a therapeutically effective amount of a composition comprising a protein of the present invention and a pharmaceutically acceptable carrier
BRIEF DESCRIPTION OF THE FIGURES
Fig 1 is an autoradiograph evidencing the expression of clones AP162, AM931 , and AR260 in COS cells (expressed band(s) indicated by dot(s)) Fig 2 is an autoradiographevidencingthe expressionofclone AM610 in COS cells
(expressed band(s) indicated by dot(s))
Fig 3 is an autoradiograph evidencing the expression of clones AM340, AM282 and AK533 in COS cells (expressed band(s) indicated by dot(s)) Fig 4 is an autoradiograph evidencing the expression of clone AK647 in COS cells (expressed band(s) indicated by dot(s))
Fig 5 is an autoradiograph evidencing the expression of clones AH583, AK296, and AS32 in COS cells (expressed band(s) indicated by dot(s)) Fig 6 is an autoradiograph evidencing the expression of clones H617 and AT205 in COS cells (expressed band(s) indicated by dot(s))
Fig 7 is an autoradiograph evidencing the expression of clones BB9 and K39 in COS cells (expressed band(s) indicated by dot(s))
Fig 8 is an autoradiograph evidencing the expression of clones AW 191 and AS34 in COS cells (expressed band(s) indicated by dot(s))
Fig 9 is an autoradiograph evidencing the expression of clones AT21 1 and AT319 in COS cells (expressed band(s) indicated by dot(s))
Fig 10 is an autoradiograph evidencing the expression of clone K640 in COS cells (expressed band(s) indicated by dot(s))
DETAILED DESCRIPTION ISOLATED PROTEINS
Nucleotide and ammo acid sequences are reported below for each clone and protem disclosed in the present application In some instances the sequences are preliminary and may include some incorrect or ambiguous bases or amino acids The actual nucleotide sequence of each clone can readily be determined by sequencing of the deposited clone m accordance with known methods The predicted ammo acid sequence (both full length and mature) can then be determined from such nucleotide sequence The amino acid sequence of the protein encoded by a particular clone can also be determined by expression of the clone in a suitable host cell, collecting the protein and determining its sequence
For each disclosed protein applicants have identified what they have determined to be the reading frame best identifiable with sequence information available at the time of filing Because of the partial ambiguity in reported sequence information, reported protem sequences include "Xaa" designators These "Xaa" designators indicate either (1) a residue which cannot be identified because of nucleotide sequence ambiguity or (2) a stop codon in the determined nucleotide sequence where applicants believe one should not exist (if the nucleotide sequence were determined definitively) 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 which are transported across the membrane of the endoplpasmic reticulum
Protein "AP162"
One protein of the present invention has been identified as protein "AP162" A partial cDNA clone encoding AP162 was first isolated from a human adult placenta cDNA library using methods which are selective for cDNAs encoding secreted proteins The nucleotide sequence of such partial cDNA was determined and searched against the GenBank database using BLASTA/BLASTX and FASTA search protocols The search revealed at least some identity with an EST reported by the I M A G E Consortium identified as "yu40d08 rl Homo sapiens cDNA clone 23671 5' " (GenBank accession number H62096) The search also found a hit at GenBank accession number H98192 The human cDNA clone corresponding to the EST database entry was ordered from Genome Systems, Inc , St Louis, Mo, a distributor of the I M A G E Consonium library The clone received from the distributor was examined and determined to be a full length clone, including a 5' end and 3' UTR (including a polyA tail) This full-length clone is also referred to herein as ΑP162"
Applicants methods identified clone API 62 as encoding a secreted protein The nucleotide sequence of the 5' portion of API 62 as presently determined is reported m SEQ ID NO 1 What applicants believe is the proper reading frame and the predicted N-terminal ammo acid sequence of the full length API 62 protem corresponding to the foregoing nucleotide sequence is reported in SEQ ID NO 2 Additional nucleotide sequence from the 3' portion of AP162, including the polyA tail, is reported in SEQ ID NO 3
Protein "AM931 " One protein of the present invention has been identified as protein "AM931 " A partial cDNA clone encoding AM931 was first isolated from a human fetal kidney cDNA library using methods which are selective for cDNAs encoding secreted proteins The nucleotide sequence of such partial cDNA was determined and searched against the GenBank database using BLASTA/BLASTX and FASTA search protocols The search revealed at least some identity with an EST reported by the I M A G E Consortium identified as "yh63e02 rl Homo sapeins cDNA clone 134426 5' " (GenBank accession number R32076) The search also found a hit at GenBank accession number N30331 The human cDNA clone corresponding to the EST database entry was ordered from Genome Systems, Inc , St Louis, Mo, a distributor of the I M A G E Consonium library The clone received from the distributor was examined and determined to be a full length clone, including a 5' end and 3' UTR (including a polyA tail) This full-length clone is also referred to herein as "AM931 "
Applicants methods identified clone AM931 as encoding a secreted protein The nucleotide sequence of AM931 as presently determined is reported in SEQ ID
NO 4 What applicants believe is the proper reading frame and the predicted ammo acid sequence of the full length AM931 protein corresponding to the foregoing nucleotide sequence is reported in SEQ ID NO 5 Amino acids 1 to 27 are the predicted leader/signal sequence, with the predicted mature amino acid sequence beginning at amino acid 28
Protein "AM610"
One protein of the present invention has been identified as protein "AM610' A partial cDNA clone encoding AM610 was first isolated from a human fetal kidney cDNA library using methods which are selective for cDNAs encoding secreted proteins The nucleotide sequence of such partial cDNA was determined and searched against the GenBank database using BLASTA/BLASTX and FASTA search protocols The search revealed at least some identity with an EST reported by the I M A G E Consortium identified as "ymOlalO rl Human EST 46249 5"* (GenBank accession number H09925) The search also found hits at GenBank accession numbers H09926 and R 14298 The human cDNA clone corresponding to the EST database entry was ordered from Genome Systems, Inc., St Louis, Mo, a distributor of the I M A G E Consonium library The clone received from the distributor was examined and determined to be a full length clone, including a 5' end and 3' UTR (including a polyA tail) This full-length clone is also referred to herein as "AM610"
Applicants' methods identified clone AM610 as encoding a secreted protein
The nucleotide sequence of the 5' portion of AM610 as presently determined is reported in SEQ ID NO 6 What applicants believe is the proper reading frame and the predicted N-terminal amino acid sequence of the full length AM610 protein corresponding
(b the foregoing nucleotide sequence is reported in SEQ ID NO 7 Ammo acids 1 to 23 are the predicted leader/signal sequence, with the predicted mature amino acid sequence beginning at ammo acid 24 Additional nucleotide sequence from the 3 ' portion of AM610, including the polyA tail, is reported in SEQ ID NO 8
Protein "AM340" One protein of the present invention has been identified as protein "AM340" A partial cDNA clone encoding AM340 was first isolated from a human fetal kidney cDNA library using methods which are selective for cDNAs encoding secreted proteins The nucleotide sequence of such partial cDNA was determined and searched against the GenBank database using BLASTA/BLASTX and FASTA search protocols The search revealed at least some identity with an EST reported by the I M A G E Consortium identified as "yo68a05 rl Homo sapiens cDNA clone 183056 5' " (GenBank accession number H42936) The search also found a hit at GenBank accession number H42872 The human cDNA clone corresponding to the EST database entry was ordered from Genome Systems, Inc , St Louis, Mo, a distributor of the I M A G E Consortium library The clone received from the distributor was examined and determined to be a full length clone, including a 5' end and 3' UTR (including a polyA tail) This full-length clone is also referred to herein as "AM340"
Applicants' methods identified clone AM340 as encoding a secreted protein The nucleotide sequence of AM340 as presently determined is reported in SEQ ID NO 9 What applicants believe is the proper reading frame and the predicted ammo acid sequence of the full length AM340 protein corresponding to the foregoing nucleotide sequence is reported in SEQ ID NO 10 Protein "AM282"
One protein of the present invention has been identified as protein "AM282" A partial cDNA clone encoding AM282 was first isolated from a human fetal kidney cDNA library using methods which are selective for cDNAs encoding secreted proteins The nucleotide sequence of such partial cDNA was determined and searched against the GenBank database using BLASTA/BLASTX and FASTA search protocols The search revealed at least some identity with an EST reported by the I M A G E Consortium identified as "yf95bl0 rl Human EST 30142 5' " (GenBank accession number R18560) The search also found a thiat GenBank accession number T96696 The human cDNA clone corresponding to the EST database entry was ordered from Genome Systems, Inc , St Louis, Mo, a distributor ot the I M A G E Consortium library The clone received from the distributor was examined and determined to be a full length clone, including a 5' end and 3' UTR (including a polyA tail) This full-length clone is also referred to herein as "AM282"
Applicants' methods identified clone AM282 as encoding a secreted protein The nucleotide sequence of the 5' poπion of AM282 as presently determined is reported in SEQ ID NO 1 1 What applicants believe is the proper reading frame and the predicted N-terminal ammo acid sequence of the full length AM282 protein corresponding to the foregoing nucleotide sequence is reported in SEQ ID NO 12 Amino acids 1 to 24 are the predicted leader/signal sequence, with the predicted mature amino acid sequence beginning at ammo acid 25 Additional nucleotide sequence from the 3' portion of AM282, including the polyA tail, is reported in SEQ ID NO 13
Protem "AK647"
One protein of the present invention has been identified as protein "AK647" A partial cDNA clone encoding AK647 was first isolated from a human fetal kidney cDNA library using methods which are selective for cDNAs encoding secreted proteins The nucleotide sequence of such partial cDNA was determined and searched against the GenBank database using BLASTA/BLASTX and FASTA search protocols The search revealed at least some identity with an EST reported by the I M A G E Consortium identified as "ym40a05 rl Human EST 50483 5' " (GenBank accession number H 17726) The human cDNA clone corresponding to the EST database entry was ordered from Genome Systems, Inc , St Louis, Mo, a distributor of the I M A G E Consortium library The clone received from the distributor was examined and determined to be a full length clone, including a 5' end and 3' UTR (including a polyA tail) This full-length clone is also referred to herein as "AK647"
Applicants' methods identified clone AK647 as encoding a secreted protein The nucleotide sequence of the 5' portion of AK647 as presently determined is reported in SEQ ID NO 14 What applicants believe is the proper reading frame and the predicted N-terminal ammo acid sequence of the full length AK647 protein corresponding to the foregoing nucleotide sequence is reported in SEQ ID NO 15 Ammo acids 1 to 25 are the predicted leader/signal sequence, with the predicted mature ammo acid sequence beginning at amino acid 26 Additional nucleotide sequence from the 3 portion of AK647, including the polyA tail, is reported in SEQ ID NO 16
Protein ΑK583"
One protein of the present invention has been identified as protein "AK583" A partial cDNA clone encoding AK583 was first isolated from a human fetal kidney cDNA library using methods which are selective for cDNAs encoding secreted proteins The nucleotide sequence of such partial cDNA was determined and searched against the GenBank database using BLASTA/BLASTX and FASTA search protocols The search revealed at least some identity with an EST reported by the I M A G E Consortium identified as τ yι90c06 rl Human EST 14656 5"' (GenBank accession number R77830) The search also found a hit at GenBank accession number H45398 The human cDN A clone corresponding to the EST database entry was ordered from Genome Systems, Inc , St Louis, Mo, a distributor of the I M A G E Consortium library The clone received from the distributor was examined and determined to be a full length clone, including a 5' end and 3' UTR (including a polyA tail) This full-length clone is also referred to herein as "AK583"
Applicants' methods identified clone AK583 as encoding a secreted protein The nucleotide sequence of the 5' portion of AK583 as presently determined is reported in SEQ ID NO 17 What applicants believe is the proper reading frame and the predicted N-terminal ammo acid sequence of the full length AK583 protein corresponding to the foregoing nucleotide sequence is reported in SEQ ID NO 18 Ammo acids 1 to 24 are the predicted leader/signal sequence, with the predicted mature ammo acid sequence beginning at ammo acid 25 Additional nucleotide sequence from the 3' portion of AK583, including the polyA tail, is reported in SEQ ID NO 19
Protem "AK533"
One protein of the present invention has been identified as protein "AK533" A partial cDNA clone encoding AK533 was first isolated from a human fetal kidney cDN A library using methods which are selective for cDNAs encoding secreted proteins The nucleotide sequence of such partial cDNA was determined and searched against the GenBank database using BLASTA/BLASTX and FASTA search protocols The search revealed at least some identity with an EST reported by the I M A G E Consortium identified as "yb82h07 rl Homo sapiens cDNA clone 77725 5' " (GenBank accession number T55939) The human cDNA clone corresponding to the EST database entry was ordered from Genome Systems, Inc , St Louis, Mo, a distributor of the I M A G E Consortium library The clone received from the distributor was examined and determined to be a full length clone, including a 5' end and 3' UTR (including a polyA tail) This full- length clone is also referred to herein as "AK533"
Applicants' methods identified clone AK533 as encoding a secreted protein The nucleotide sequence of the 5' portion of AK533 as presently determined is reported m SEQ ID NO 20 What applicants believe is the proper reading frame and the predicted N-terminal amino acid sequence of the full length AK533 protein corresponding to the foregoing nucleotide sequence is reported in SEQ ID NO 21 Additional nucleotide sequence from the 3' portion of AK533, including the polyA tail, is reported in SEQ ID NO 22
Protein "AK296"
One protein of the present invention has been identified as protein "AK296" A partial cDNA clone encoding AK296 was first isolated from a human fetal kidney cDNA library using methods which are selective for cDNAs encoding secreted proteins The nucleotide sequence of such partial cDNA was determined and searched against the GenBank database using BLASTA/BLASTX and FASTA search protocols The search revealed at least some identity with an EST reported by the I M A G E Consortium identified as "yc86gl2 rl Homo sapeins cDNA clone 22958 5' " (GenBank accession number T75226) The human cDNA clone corresponding to the EST database entry was ordered from Genome Systems, Inc , St Louis, Mo, a distributor of the I M A G E Consortium library The clone received from the distributor was examined and determined to be a full length clone, including a 5' end and 3' UTR (including a polyA tail) This full- length clone is also referred to herein as ' AK296" Applicants' methods identified clone AK296 as encoding a secreted protein
The nucleotide sequence of the 5 portion of AK296 as presently determined is reported in SEQ ID NO 23 What applicants believe is the proper reading frame and the predicted N-terminal amino acid sequence of the full length AK296 protein corresponding to the foregoing nucleotide sequence is reported in SEQ ID NO 24 Ammo acids 1 to 36 are the predicted leader/signal sequence, with the predicted mature amino acid sequence beginning at ammo acid 37 Additional nucleotide sequence from the 3 ' portion of AK296, including the polyA tail, is reported m SEQ ID NO 25
Protein "H617"
One protein of the present invention has been identified as protein "H617" A partial cDNA clone encoding H617 was first isolated from a human PBMC cDNA library using methods which are selective for cDNAs encoding secreted proteins The nucleotide sequence of such partial cDN A was determined and searched against the GenBank database using BLASTA/BLASTX and FASTA search protocols The search revealed at least some identity with an EST reported by the I M A G E Consortium identified as "ysl lcl2 rl Homo sapeins cDNA clone 214486 5' " (GenBank accession number H71514) The search also found a hit at GenBank accession number R10010 The human cDNA clone corresponding to the EST database entry was ordered from Genome Systems, Inc , St
Louis, Mo, a distributor of the I M A G E Consortium library The clone received from the distributor was examined and determined to be a full length clone, including a 5' end and 3' UTR (including a polyA tail) This full-length clone is also referred to herein as "H617" Applicants' methods identified clone H617 as encoding a secreted protein
The nucleotide sequence of the 5' portion of H617 as presently determined is reported in SEQ ID NO 26 What applicants believe is the proper reading frame and the predicted N-terminal amino acid sequence of the full length H617 protein corresponding to the foregoing nucleotide sequence is reported in SEQ ID NO 27 Additional nucleotide sequence from the 3' portion of H617, including the polyA tail, is reported in SEQ ID
KI0 28
Protem "BB9"
One protein of the present invention has been identified as protein "BB9" A partial cDNA clone encoding BB9 was first isolated from a human PBMC (TH 1 or Th2) cDNA library using methods which are selective for cDNAs encoding secreted proteins The nucleotide sequence of such partial cDNA was determined and searched against the GenBank database using BLASTA/BLASTX and FASTA search protocols The search revealed at least some identity with an EST reported by the I M A G E Consortium identified as "yd68g04 rl Human cDNA clone 113430 5' " (GenBank accession number T78562) The search also found a thi at GenBank accession number R54388 The human cDNA clone corresponding to the EST database entry was ordered from Genome Systems,
Inc , St Louis, Mo, a distributor of the I M A G E Consortium library The clone received from the distributor was examined and determined to be a full length clone, including a 5 end and 3' UTR (including a polyA tail) This full-length clone is also referred to herein as "BB9" Applicants' methods identified clone BB9 as encoding a secreted protein
The nucleotide sequence of the 5' portion of BB9 as presently determined is reported in SEQ ID NO 29 What applicants believe is the proper readmg frame and the predicted N-terminal amino acid sequence of the full length BB9 protein corresponding to the foregoing nucleotide sequence is reported in SEQ ID NO 30 Additional nucleotide sequence from the 3' portion of BB9, including the polyA tail, is reported in SEQ ID
NO.31. Protein ΑW191 "
One protein of the present invention has been identified as protein "AW191 " A partial cDNA clone encoding AW 191 was first isolated from a human ovary (PA-1 teratocarcmoma) cDNA library using methods which are selective for cDNAs encoding secreted proteins The nucleotide sequence of such partial cDNA was determined and searched against the GenBank database using BLASTA/BLASTX and FASTA search protocols The search revealed at least some identity with an EST reported by the I M.A G E Consortium identified as "ym03dl0 rl Homo sapiens cDNA clone 46942 5' " (GenBank accession number H 10314 The search also found a hit at GenBank accession number H05460 The human cDN A clone corresponding to the EST database entry was ordered from Genome Systems, Inc , St Louis, Mo, a distributor of the I M A G E Consortium library The clone received from the distributor was examined and determined to be a full length clone, including a 5' end and 3' UTR (including a polyA tail) This full- length clone is also referred to herein as 'AW191 " Applicants' methods identified clone AW191 as encoding a secreted protein
The nucleotide sequence of the 5' portion of AW 191 as presently determined is reported in SEQ ID NO 32 What applicants believe is the proper reading frame and the predicted N-terminal amino acid sequence of the full length AW 191 protein corresponding to the foregoing nucleotide sequence is reported m SEQ ID NO 33 Ammo acids 1 to 18 are the predicted leader/signal sequence, with the predicted mature amino acid sequence beginning at ammo acid 19 Additional nucleotide sequence from the 3 ' portion of AW 191, including the polyA tail, is reported in SEQ ID NO 34
Protem "AT2U "
One protein of the present invention has been identified as protein "AT21 1 " A partial cDNA clone encoding AT2I 1 was first isolated from a human lymphocyte and dendritic cell cDNA library using methods which are selective for cDNAs encoding secreted proteins The nucleotide sequence of such partial cDNA was determined and searched against the GenBank database using BLASTA/BLASTX and FASTA search protocols The search revealed at least some identity with an EST reported by the I M A G.E Consortium identified as "yq36f01 rl Homo sapiens cDNA clone 197881 5'" (GenBank accession number R96278) The search also found a hit at GenBank accession number R56077 The human cDNA clone corresponding to the EST database entry was ordered from Genome Systems, Inc , St Louis, Mo, a distributor of the I M A G E Consortium library The clone received from the distributor was examined and determined to be a full length clone, including a 5' end and 3' UTR (including a polyA tail) This full- length clone is also referred to herein as "AT211 "
Applicants' methods identified clone AT211 as encoding a secreted protein The nucleotide sequence of the 5' portion of AT21 1 as presently determined is reported in SEQ ID NO 35 What applicants believe is the proper reading frame and the predicted N-termmal amino acid sequence of the full length AT211 protein corresponding to the foregoing nucleotide sequence is reported in SEQ ID NO 36 Ammo acids 1 to 53 are the predicted leader/signal sequence, with the predicted mature ammo acid sequence beginning at ammo acid 54 Additional nucleotide sequence from the 3 ' portion of AT211 , including the polyA tail, is reported in SEQ ID NO 37
Protem "AT205"
One protein of the present invention has been identified as protein "AT205" A partial cDNA clone encoding AT205 was first isolated from a human lymphocyte and dendritic cell cDNA library using methods which are selective for cDNAs encoding secreted proteins The nucleotide sequence of such partial cDNA was determined and searched against the GenBank database using BLASTA/BLASTX and FASTA search protocols The search revealed at least some identity with an EST reported by the I M A G E Consortium identified as 'yu83c l 1 rl Homo sapiens cDNA clone 240404 5" (GenBank accession number H78080) The search also found a hit at GenBank accession number H78081 The human cDN A clone corresponding to the EST database entry was ordered from Genome Systems, Inc , St Louis, Mo, a distributor of the I M A G E Consortium library The clone received from the distributor was examined and determined to be a full length clone, including a 5' end and 3' UTR (including a polyA tail) This full- length clone is also referred to herein as "AT205"
Applicants' methods identified clone AT205 as encoding a secreted protein The nucleotide sequence of AT205 as presently determined is reported in SEQ ID NO 38 What applicants believe is the proper reading frame and the predicted amino acid sequence of the full length AT205 protein corresponding to the foregoing nucleotide sequence is reported in SEQ ID NO 39 Ammo acids 1 to 55 are the predicted leader/signal sequence, with the predicted mature amino acid sequence beginning at amino acid 56
Protein "AS34"
One protem of the present invention has been identified as protein "AS34" A partial cDNA clone encoding AS34 was first isolated from a human fetal brain cDNA library using methods which are selective for cDNAs encoding secreted proteins The nucleotide sequence of such partial cDNA was determined and searched against the GenBank database using BLASTA/BLASTX and FASTA search protocols The search revealed at least some identity with an EST reported by the I M A G E Consortium identified as "yg71a01 rl Homo sapiens cDNA clone 38531 5' " (GenBank accession number R51 1 18) The search also found a hit at GenBank accession number R 15801 The human cDNA clone corresponding to the EST database entry was ordered from Genome Systems, Inc , St Louis, Mo, a distributor of the I M A G E Consortium library The clone received from the distributor was examined and determined to be a full length clone, including a 5' end and 3' UTR (including a polyA tail) This full-length clone ts also referred to herein as "AS34" Applicants' methods identified clone AS34 as encoding a secreted protein
The nucleotide sequence of the 5' portion of AS34 as presently determined is reported in SEQ ID NO 40 What applicants believe is the proper reading frame and the predicted N-terminal amino acid sequence of the full length AS34 protein corresponding to the foregoing nucleotide sequence is reported in SEQ ID NO 41 Ammo acids 1 to 24 are the predicted leader/signal sequence, with the predicted mature amino acid sequence beginning at ammo acid 25 Addinonal nucleotide sequence from the 3 portion of AS34, including the polyA tail, is reported in SEQ ID NO 42
Protem "AS32"
One protein of the present invention has been identified as protein "AS32" A partial cDNA clone encoding AS32 was first isolated from a human fetal brain cDNA library using methods which are selective for cDNAs encoding secreted proteins The nucleotide sequence of such partial cDNA was determined and searched against the GenBank database using BLASTA/BLASTX and FASTA search protocols The search revealed at least some identity with an EST reported by the I M A G E Consortium identified as "yu75b08 rl Homo sapiens cDNA clone 239607 5' " (GenBank accession number H80466) The search also found a hit at GenBank accession number H77627 The human cDNA clone corresponding to the EST database entry was ordered from Genome Systems, Inc , St Louis, Mo, a distributor of the I M A G E Consortium library The clone received from the distributor was examined and determined to be a full length clone, including a 5' end and 3' UTR (including a polyA tail) This full-length clone is also referred to herein as "AS32"
Applicants' methods identified clone AS32 as encoding a secreted protein The nucleotide sequence of the 5 portion of AS32 as presently determined is reported in SEQ ID NO 43 What applicants believe is the proper reading frame and the predicted N-terminal amino acid sequence of the full length AS32 protein corresponding to the foregoing nucleotide sequence is reported in SEQ ID NO 44 Additional nucleotide sequence from the 3' portion of AS32, including the polyA tail, is reported in SEQ ID NO 45
Protein "AR260"
One protein of the present invention has been identified as protein ΑR260" A partial cDNA clone encoding AR260 was first isolated from a human adult retina cDNA library using methods which are selective for cDNAs encoding secreted proteins The nucleotide sequence of such partial cDNA was determined and searched against the
GenBank database using BLASTA/BLASTX and FASTA search protocols The search revealed at least some identity with an EST reported by the I M A G E Consortium identified as "yg99gl2 rl Homo sapiens cDNA clone 41757 5' " (GenBank accession number R52804) The human cDNA clone corresponding to the EST database entry was ordered from Genome Systems, Inc , St Louis, Mo, a distributor of the I M A G E
Consortium library The clone received from the distributor was examined and determined to be a full length clone, including a 5' end and 3' UTR (including a polyA tail) This full- length clone is also referred to herein as "AR260"
Applicants' methods identified clone AR260 as encoding a secreted protein The nucleotide sequence of the 5 portion of AR260 as presently determined is reported in SEQ ID NO 46 What applicants believe is the proper reading frame and the predicted N-terminal ammo acid sequence of the full length AR260 protein corresponding to the foregoing nucleotide sequence is reported in SEQ ID NO 47 Amino acids 1 to 23 are the predicted leader/signal sequence, with the predicted mature ammo acid sequence beginning at ammo acid 24 Additional nucleotide sequence from the 3 portion of AR260, mcluding the polyA tail, is reported in SEQ ID NO 48
Protein ' K640"
One protein of the present invention has been identified as protein "K640 ' A partial cDNA clone encoding K640 was first isolated from a murine bone marrow (stromal cell line FCM-4) cDNA library using methods which are selective for cDNAs encoding secreted proteins The nucleotide sequence of such partial cDN A was determined and searched against the GenBank database using BLASTA/BLASTX and FASTA search protocols The search revealed at least some identity with an EST reported by the I M A G E Consortium identified as 'yf47a09 rl Homo sapiens cDNA clone 129976 5' " (GenBank accession number Rl 1595) The search also found a hit at GenBank accession number H09031 The human cDNA clone corresponding to the EST database entry was ordered from Genome Systems, Inc , St Louis, Mo, a distributor of the I M A G E Consortium library The clone received from the distributor was examined and determined to be a full length clone, including a 5 end and 3 ' UTR (including a polyA tail) This full- length clone is also referred to herein as 'K640" Applicants' methods identified clone K640 as encoding a secreted protem
The nucleotide sequence of the 5' portion of K640 as presently determined is reported in SEQ ID NO 49 What applicants believe is the proper reading frame and the predicted N-terminal amino acid sequence of the full length K640 protein corresponding to the foregoing nucleotide sequence is reported in SEQ ID NO 50 Additional nucleotide sequence from the 3' portion of K640, including the polyA tail, is reported in SEQ'ID
NO 51 Protein "K39"
One protein of the present invention has been identified as protein "K39" . A partial cDNA clone encoding K39 was first isolated from a murine bone marrow (stromal cell line FCM-4) cDNA library using methods which are selective for cDNAs encoding secreted proteins. The nucleotide sequence of such partial cDNA was determined and searched against the GenBank database using BLASTA/BLASTX and FASTA search protocols. The starch revealed at least some identity with an EST reported by the I.M.A.G.E. Consortium identified as "ym65b04.rl Homo sapiens cDNA clone 163759 5' " (GenBank accession number H 14129). The search also found a hit at GenBank accession number H68304. The human cDN A clone corresponding to the EST database entry was ordered from Genome Systems, Inc. , St. Louis, Mo, a distributor of the I.M.A.G.E. Consortium library. The clone received from the distributor was examined and determined to be a full length clone, including a 5' end and 3' UTR (including a polyA tail). This full-length clone is also referred to herein as "K39" . Applicants' methods identified clone K39 as encoding a secreted protein.
The nucleotide sequence of the 5' portion of K39 as presently determined is reported in SEQ ID NO:52. What applicants believe is the proper reading frame and the predicted N-terminal amino acid sequence of the full length K39 protein corresponding to the foregoing nucleotide sequence is reported in SEQ ID NO:53. Additional nucleotide sequence from the 3' portion of K39, including the polyA tail, is reported in SEQ ID NO:54.
Protein "AT319"
One protein of the present invention has been identified as protein "AT319". A partial cDNA clone encoding AT319 was first isolated from a human lymphocyte and dendritic cell cDNA library using methods which are selective for cDNAs encoding secreted proteins . The nucleotide sequence of such partial cDN A was determined and searched against the GenBank database using BLASTA/BLASTX and FASTA search protocols. The search revealed at least some identity with an EST reported by the I.M.A.G.E. Consortium identified as "yr21bl l .rl Homo sapiens cDNA clone 205917 5'" (GenBank accession number H57730). The search also found a hit at GenBank accession number H57731. The human cDN A clone corresponding to the EST database entry was ordered from Genome Systems, Inc , St Louis. Mo, a distributor of the I M A.G E Consortium library The clone received from the distributor was examined and determined to be a full length clone, including a 5' end and 3' UTR (including a polyA tail) This full- length clone is also referred to herein as "AT319" Applicants' methods identified clone AT319 as encoding a secreted protein
The nucleotide sequence of the 5' portion of AT319 as presently determined is Reported in SEQ ID N0 55 What applicants believe is the proper reading frame and the predicted N-terminai amino acid sequence of the full length AT319 protein corresponding to the foregoing nucleotide sequence is reported in SEQ ID NO:56 Additional nucleotide sequence from the 3' portion of AT319, including the polyA tail, is reported in SEQ ID NO:57
Deposit of Clones
Clones AP162, AM931 , AM610, AM340, AM282, AK647. AK583. AK533, AK296, H617, BB9, AW191 , AT211 , AT205, AS34, AS32, AR260, K640, K39 and AT319 were deposited on April 17, 1996 with the American Type Culture Collection under accession number ATCC 98026, from which each clone comprising a particular polynucleotide is obtainable Each clone has been transfected into separate bacterial cells (E colt) m this composite deposit 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 design of the oligonucleotide probe should preferably follow these parameters (a) It should be designed to an area of the sequence which has the fewest ambiguous bases ("N's"), if any, (b) It should be designed to have a Tm of approx 80 ° C (assuming 2° for each
A or T and 4 degrees for each G or C) The oligonucleotide should preferably be labeled with g-32P ATP (specific activity 6000 Ci/mmole) and T4 polynucleotide kinase using commonly employed techniques for labeling oligonucleotides Other labeling techniques can also be used Unincorporated label should preferably be removed by gel filtration chromatography or other established methods The amount of radioactivity incoφorated into the probe should be quantitated by measurement in a scintillation counter Preferably, specific activity of the resulting probe should be approximately 4e + 6 dpm/pmole
The bacterial culture containing the pool of full-length clones should preferably be thawed and 100 μl of the stock used to inoculate a sterile culture flask containing 25 ml of sterile L-broth containing ampicillin at 100 μg/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 μg/ml and agar at 1 5% in a 150 mm petri dish when grown overnight at 37°C Other known methods of obtaining distinct, well-separated colonies can also be employed
Standard colony hybridization procedures should then be used to transfer the colonies to nitrocellulose filters and lyse, denature and bake them The filter is then preferably incubated at 65 °C for 1 hour with gentle agitation in 6X SSC (20X stock is 175.3 g NaCI/liter, 88.2 g Na citrate/liter, adjusted to pH 7.0 with NaOH) containing 0.5% SDS, 100 μg/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 0.1X SSC/0.5 % SDS at 65 °C for 30 minutes to 1 hour is optional. The filter is then preferably dried and subjected to autoradiography for sufficient time to visualize the positives on the X-ray film. Other known hybridization methods can also be employed.
The positive colonies are picked, grown in culture, and plasmid DNA isolated using standard procedures. The clones can then be verified by restriction analysis, hybridization analysis, or DNA sequencing.
Fragments of the proteins of the present invention which are capable of exhibiting biological activity are also encompassed by the present invention. Fragments of the protein may be in linear form or they may be cyclized using known methods, for example, as described in H.U. Saragovi, et al. , Bio/Technology 10, 773-778 (1992) and in R.S. McDowell, et al. , J. Amer. Chem. Soc. J_14, 9245-9253 ( 1992), both of which are incoφorated herein by reference. Such fragments may be fused to carrier molecules such as immunoglobulins for many puφoses, 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 form 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 sequence of the mature form of the protein may also be determinable from the amino acid sequence of the full-length form.
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 pan or ail of the intracellular and transmembrane domains of the protein are deleted such that the protein is fully secreted from the cell in which it is expressed. The intracellular and fransmembrane domains of proteins of the invention can be identified in accordance with known techniques for determination of such domains from sequence information.
Species homologs of the disclosed proteins are also provided by the present invention. Species homologs 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.
The invention also encompasses allelic variants of the disclosed proteins; that is, naturally-occurring alternative forms of the isolated proteins which are identical, homologous or related to that encoded by the polynucleotides disclosed herein.
The isolated polynucleotide endcoing the protein of the invention may be operably linked to an expression control sequence such as the pMT2 or pED expression vectors disclosed in Kaufman et al.. Nucleic Acids Res. __, 4485-4490 (1991), in order to produce the protein recombinantly. Many suitable expression control sequences are known in the art. General methods of expressing recombinant proteins are also known and are exemplified in R. Kaufman, Methods in Enzymology 185. 537-566 (1990). As defined herein "operably 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 protein. Mammalian host cells include, for example, monkey COS cells, Chinese Hamster Ovary (CHO) cells, human kidney 293 cells, human epidermal A431 cells, human Colo205 cells, 3T3 cells, CV-1 cells, other transformed primate cell lines, normal diploid cells, cell strains derived from in vitro culture of primary tissue, primary explants, HeLa cells, mouse
L cells, BHK, HL-60, U937, HaK or Jurkat cells.
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, Schizosaccharomycβ pombe, Kluyveromyces strains, Candida, or any yeast strain capable of expressing heterologous proteins Potentially suitable bacterial strains include Escherichia colt, Bacillus subtilis, Salmonella rvphtmurium, 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, m order to obtain the functional protein Such covalent attachments may be accomplished using known chemical or enzymatic methods
The protein may also be produced by operably linking the isolated polynucleotide of the invention to suitable control sequences in one or more insect expression vectors, and employing an insect expression system Materials and methods for baculovirus/insect cell expression systems are commercially available in kit form from, e g , Invitrogen, San Diego, California, U S A (the MaxBac® kit), and such methods are well known in the art, as described in Summers and Smith, Texas Agricultural Experiment Station Bulletin No 1555 (1987). incoφorated 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 containing agents which will bind to the protein, one or more column steps over such affinity resins as concanavalin A-agarose, hepaπn-toyopearl® or Cibacrom blue 3GA Sepharose®, one or more steps involving hydrophobic interaction chromatography using such resins as phenyl ether, butyl ether, or propyl ether, or immunoaffinity chromatography
Alternatively, the protein of the invention may also be expressed in a form which will facilitate purification For example, it may be expressed as a fusion protein, such as those of maltose binding protein (MBP), glutathione-S-transferase (GST) or thioredoxin (TRX) Kits for expression and purification of such fusion proteins are commercially available from New England BioLab (Beverly, MA), Pharmacia (Piscataway, NJ) and
InVitrogen, 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 Kodak (New Haven, CT) Finally, one or more reverse-phase high performance liquid chromatography (RP- HPLC) steps employing hydrophobic RP-HPLC media, e g , sihca 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 tne 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-constructedprotein sequences, by virtue of sharing primary, secondary or tertiary structural and/or conformational characteristics with proteins may possess biological properties in common therewith, including protein activity Thus, they may be employed as biologically active or immunological substitutes for natural, purified proteins in screening of therapeutic compounds and in immunological processes for the development of antibodies The proteins provided herein also include proteins characteπzed 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, insertion or deletion of a selected ammo 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, 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 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 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-hgand 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 bmding 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 Molecular
Cloning Techniques", Academic Press, Berger, S L and A R Kimmel eds , 1987
Nmπnonal Uses Proteins of the present invention can also be used as nutritional sources or supplements . Such uses include without limitation use as a protein 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 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 of the invention can be added to the medium in or on which the microorganism is cultured.
Cytokine and Cell Proliferation/Differentiation Activity
A protein of the present invention may exhibit cytokine, cell proliferation (either inducing or inhibiting) or cell differentiation (either inducing or inhibiting) activity or may 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, B9/1 1 , BaF3, MC9/G, M + (preB M + ), 2E8, RB5, DAI , 123, T1165, HT2. CTLL2, TF-1 , Mo7e and CMK. The activity of a protein of the invention may, among other means, be measured by the following methods:
Assays for T-cell or thymocyte proliferation include without limitation those described in: Current Protocols in Immunology, Ed by J. E. Coligan, A.M. Kruisbeek, D.H. Margulies, E.M. Shevach, W Strober, Pub. Greene Publishing Associates and Wiley- Interscience (Chapter 3, In Vitro assays for Mouse Lymphocyte Function 3.1-3.19; Chapter 7, Immunologic studies in Humans); Takai et al., J. Immunol. 137:3494-3500, 1986; Bertagnolli et al., J. Immunol. 145: 1706-1712, 1990; Bertagnolli et al., Cellular Immunology 133:327-341, 1991 ; Bertagnolli, et al., J. Immunol. 149:3778-3783, 1992; Bowman et al., J. Immunol. 152: 1756-1761 , 1994. Assays for cytokine production and/or proliferation of spleen cells, lymph node 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 γ, Schreiber, R.D. In Current Protocols in Immunology J E e a Coligan eds Vol 1 pp 6 8 1-6 8 8, John Wiley and Sons, Toronto 1994
Assays for prohferationand differentiationof hematopoieticand lymphopoietic cells include, without limitation, those described in Measurement of Human and Murine Interieukin 2 and Interieukin 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 , deVπes et al , J Exp Med 173 1205-1211, 1991, Moreau et al , Nature 336 690-692, 1988, Greenberger et al , Proc Natl Acad Sci U S A 80 2931-2938, 1983, Measurement of mouse and human interieukin 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 Interieukin 1 1 - Bennett, F , Giannotti, J , Clark, S C and Turner, K J In Current Protocols in Immunology J E e a Coligan eds Vol 1 pp 6 15 1 John Wiley and Sons, Toronto 1991 , Measurement of mouse and human Interieukin 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 Marguiies, E M Shevach, W Strober, Pub Greene Publishing Associates and Wiley-Interscience (Chapter 3, In Vitro assays for Mouse Lymphocyte Function, Chapter 6, Cytokines and their cellular receptors. Chapter 7, Immunologic studies in Humans), Weinberger et al , Proc Natl Acad Sci USA 77 6091-6095, 1980, Weinberger et al , Eur J Immun 11 405-411 , 1981 , Takai et al , J Immunol 137 3494-3500, 1986, Takai et al , J
Immunol 140 508-512, 1988
Immune Stimulating or Suppressing Activity
A protein of the present invention may also exhibit immune stimulating or immune suppressing activity, including without limitation the activities for which assays are described herein A protein may be useful in the treatment of various immune deficiencies and disorders (including severe combined immunodeficiency (SCID)), e g , in regulating (up or down) growth and proliferation of T and/or B lymphocytes, as well as effecting the cytolytc activity of NK cells and other cell populations These immune deficiencies may be genetic or be caused by viral (e.g , HIV) as well as bacterial or fungal infections, or may result from autoimmune disorders More specifically, infectious diseases causes by viral, bacterial , fungal or other infection may be treatable using a protein of the present invention, including infections by HIV, hepatitis viruses, heφesviruses, mycobacteπa, 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, / e , in the treatment of cancer
Autoimmune disorders which may be treated using a protein of the present invention include, for example, connective tissue disease, multiple sclerosis, systemic lupus erythematosus, rheumatoid arthritis, autoimmune pulmonary inflammation, Guillain-Barre syndrome, autoimmune thyroiditis, insulin dependent diabetes melhtis, myasthenia gravis, graft-versus-hostdisease 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 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 toleπzing 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 toleπzing agent Down regulating or preventing one or more antigen functions (including without limitation B lymphocyte antigen functions (such as , for example, B7)), e g , preventing high level lymphokme synthesis by activated T cells, will be useful in situations of tissue, skm and organ transplantation and in graft-versus-host disease (GVHD) For example, blockage of T cell function should result in reduced tissue destruction in tissue transplantation Typically, in tissue transplants, rejection of the transplant is initiated through its recognition as foreign by T cells, followed by an immune reaction that destroys the transplant The administration of a molecule which inhibits or blocks interaction of a B7 lymphocyte antigen with its natural hgand(s) on immune cells (such as a soluble, monomeric form of a peptide having B7-2 activity alone or in conjunction with a monomeric form of a peptide having an activity of another B lymphocyte antigen (e g , B7- 1 , B7-3) or blocking antibody), prior to transplantation can lead to the binding of the molecule to the natural lιgand(s) on the immune cells without transmitting the corresponding dostimulatory 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 costimulationmay also be sufficient to anergize the T cells, thereby inducing tolerance in a subject Induction of long-term tolerance by B lymphocyte antigen- blocking reagents may avoid the necessity of repeated administration of these blocking reagents To achieve sufficient immunosuppression or tolerance in a subject, it may also be necessary to block the function of a combination of B lymphocyte antigens
The efficacy of particular blocking reagents in preventing organ transplant rejection or GVHD can be assessed using animal models that are predictive of efficacy in humans Examples of appropriate systems which can be used include allogeneic cardiac grafts in rats and xenogeneic pancreatic islet cell grafts in mice, both of which have been used to examine the immunosuppressive effects of CTLA4Ig fusion proteins in vivo as described in Lenschow et al , Science 257789-792 (1992) and Turka et al , Proc Natl. Acad Sci USA, 89 1 1102-11105 (1992) In addition, murine models of GVHD (see Paul ed , Fundamental Immunology, Raven Press, New York, 1989, pp 846-847) can be used to determine the effect of blocking B lymphocyte antigen function in vivo on the development of that disease
Blocking antigen function may also be therapeutically useful for treating autoimmune diseases Many autoimmune disorders are the result of inappropriate activation of T cells that are reactive against self tissue and which promote the production of cytokines and autoantibodes involved in the pathology of the diseases Preventing the 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 m the disease process Additionally, blocking reagents may induce antigen-specific tolerance of autoreactive T cells which could lead to long-term relief from the disease. The efficacy of blocking reagents in preventing or alleviating autoimmune disorders can be determined using a number of well-characterized animal models of human autoimmune diseases. Examples include murine experimental autoimmune encephalitis, systemic lupus erythmatosis in MRL/lpr/lpr mice or NZB hybrid mice, murine autoimmune collagen arthritis, diabetes mellitus in NOD mice and BB rats, and murine 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), a's a means of up regulating immune responses, may also be useful in therapy. Upregulation of immune responses may be in the form of enhancing an existing immune response or eliciting an initial immune response. For example, enhancing an immune response through stimulating B lymphocyte antigen function may be useful in cases of viral infection. In addition, systemic viral diseases such as influenza, the common cold, and encephalitis might be alleviated by the administration of stimulatory forms of B lymphocyte antigens systemically.
Alternatively, anti-viral immune responses may be enhanced in an infected patient by removing T cells from the patient, costimulating the T cells in vitro with viral antigen- pulsed APCs either expressing a peptide of the present invention or together with a stimulatory form of a soluble peptide of the present invention and reintroducing the in vitro activated T cells into the patient. Another method of enhancing anti-viral immune responses would be to isolate infected cells from a patient, transfect them with a nucleic acid encoding a protein of the present invention as described herein such that the cells express all or a portion of the protein on their surface, and reintroduce the transfected cells into the patient. The infected cells would now be capable of delivering a costimulatory signal to, and thereby activate, T cells in vivo.
In another application, up regulation or enhancement of antigen function (preferably B lymphocyte antigen function) may be usefiil in the induction of tumor immunity. Tumor cells (e.g., sarcoma, melanoma, lymphoma, leukemia, neuroblastoma, carcinoma) transfected with a nucleic acid encoding at least one peptide of the present invention can be administered to a subject to overcome tumor-specific tolerance in the subject. If desired, the tumor cell can be transfected to express a combination of peptides . For example, tumor cells obtained from a patient can be transfected ex vivo with an expression vector directing the expression of a peptide having B7-2-like activity alone, or in conjunction with a peptide having B7-l-like activity and/or B7-3-like activity. The transfected tumor cells are returned to the patient to result in expression of the peptides on the surface of the transfected cell Alternatively, gene therapy techniques can be used to target a tumor cell for transfection in vivo
The presence of the peptide of the present invention having the activity of a B lymphocyte antιgen(s) on the surface of the tumor cell provides the necessary costimulation signal to T cells to induce a T cell mediated immune response against the transfected tumor cells In addition, tumor cells which lack MHC class I or MHC class II molecules, or which fail to reexpress sufficient amounts of MHC class I or MHC class II molecules, can be transfected with nucleic acid encoding all or a portion of (e g , a cytoplasmic-domain truncated portion) of an MHC class I α chain protein and β2 microglobuhn protein or an MHC class II a chain protein and an MHC class II β 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-l, 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-spec if ic 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 limitatioα those described in Current Protocols in Immunology, Ed by J E Coligan, A M Kruisbeek, D H. Marguiies, E M Shevach, W Strober, Pub Greene Publishing Associates and Wiley-Interscience (Chapter 3, In Vitro assays for Mouse Lymphocyte Function 3 1- 3.19; Chapter 7, Immunologic studies in Humans); Herrmann et al., Proc Natl Acad. Sci USA 78.2488-2492 , 1981 ; Herrmann et al . , J . Immunol 128.1968- 1974 , 1982 , Handa et al , 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. Immunol 137 3494-3500, 1986, Bowmanet al., J 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 in Mahszewski, 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, those described in Current Protocols in Immunology, Ed by J E Coligan, A M Kruisbeek, D H Marguiies, 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 m Darzynkiewicz et al , Cytometry 13 795-S08, 1992, Gorczyca et al , Leukemia 7 659-670, 1993, Gorczyca et al , Cancer Research 53 1945-1951 , 1993, Itoh et al , Cell 66 233-243, 1991 , Zacharchuk, Journal of Immunology 145 4037-4045, 1990, Zamai et al , Cytometry 14 891-897, 1993, 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 11 1-117, 1994, Fine et al , Cellular Immunology 155 1 11-122, 1994, Galy et al , Blood
85 2770-2778, 1995, Toki et al , Proc Nat Acad Sci USA 88 7548-7551 , 1991 Hematopoiesis Regulating Activity
A protein of the present invention may be useful in regulation of hematopoiesis and, consequently, in the treatment of myeloid or lymphoid cell deficiencies. Even marginal biological activity in support of colony forming cells or of factor-dependent cell 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, (hereby indicating utility, for example, in treating various anemias or for use in conjunction with irradiation/chemotherapy to stimulate the production of erythroid precursors and/or erythroid cells; in supporting the growth and proliferation of myeloid cells such as granulocytes and monocytes/macrophages (i.e., traditional CSF activity) useful, for example, in conjunction with chemotherapy to prevent or treat consequent myelo¬ suppression; in supporting the growth and proliferation of megakaryocytesand consequently of platelets thereby allowing prevention or treatment of various platelet disorders such as thrombocytopenia, and generally for use in place of or complimentary to platelet transfusions; and/or in supporting the growth and proliferation of hematopoietic stem cells which are capable of maturing to any and all of the above-mentioned hematopoietic cells and therefore find therapeutic utility in various stem cell disorders (such as those usually treated with transplantation, including, without limitation, aplastic anemia and paroxysmal nocturnal hemoglobinuria), as well as in repopulating the stem cell compartment post irradiation/chemotherapy, either in-vivo or ex-vivo (i.e., in conjunction with bone marrow transplantation or with peripheral progenitor cell transplantation (homologous or heterologous)) as normal cells or genetically manipulated for gene therapy.
The activity of a protein of the invention may, among other means, be measured by the following methods: Suitable assays for proliferation and differentiation of various hematopoietic lines are cited above.
Assays for embryonic stem cell differentiation (which will identify, among others, proteins that influence embryonic differentiation hematopoiesis) include, without limitation, those described in: Johansson et al. Cellular Biology 15: 141-151 , 1995; Keller et al., Molecular and Cellular Biology 13:473-486, 1993; McClanahan et al.. Blood 81 :2903-2915, 1993.
Assays for stem cell survival and differentiation (which will identify, among 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 89:5907-591 1 , 1992; Primitive hematopoietc colony forming cells with high proliferative potential, McNiece, I K. and Briddell, R.A. In Culture of Hematopoietic Cells. R.I. Freshney, et al. eds. Vol pp. 23-39, Wiley-Liss, Inc. , New York, NY. 1994; Neben et al., Experimental Hematology 22:353-359, 1994; Cobblestone area forming cell assay, Ploemacher, R.E. In Culture of Hematopoietic Cells. R.I. Freshney, et al. eds. Vol pp. 1-21 , Wiley-Liss, Inc . , New York, NY. 1994; Long term bone marrow cultures in the presence of stromal cells, Spooncer, E. , Dexter, M. and Allen, T. In Culture of Hematopoietic Cells. R.I. Freshney, et al. eds. Vol pp. 163-179, Wiley-Liss, Inc., New York, NY. 1994; Long term culture initiating cell assay, Sutherland, H.J. In Culture of Hematopoietic Cells. R.I. Freshney, et al. eds. Vol pp. 139-162, Wiley-Liss, Inc. , New York, NY. 1994.
Tissue Growth Activity A protein of the present invention also may have utility in compositions used for bone, cartilage, tendon, ligament and/or nerve tissue growth or regeneration, as well as for wound healing and tissue repair and replacement, and in the treatment of burns, incisions and ulcers.
A protein of the present invention, which induces cartilage and/or bone growth in circumstances where bone is not normally formed, has application in the healing of bone fractures and cartilage damage or defects in humans and other animals. Such a preparation employing a protein of the invention may have prophylactic use in closed as well as open fracture reduction and also in the improved fixation of artificial joints. De novo bone formation induced by an osteogenic agent contributes to the repair of congenital, trauma induced, or oncologic resection induced craniofacial defects, and also is useful in cosmetic plastic surgery.
A protein of this invention may also be used in the treatment of periodontal 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 invention, which induces tendon/ligament-like tissue or other tissue formation in circumstances where such tissue is not normally formed, has application in the healing of tendon or ligament tears, deformities and other tendon or ligament defects in humans and other animals. Such a preparation employing a tendon/ligament-liketissue 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-liketissue formation induced by a composition of the present invention contributes to the repair of congenital, trauma induced, or other tendon or ligament defects of other origin, and is also useful in cosmetic plastic surgery for attachment or repair of tendons or ligaments. The compositions of the present invention may provide an environment to attract tendon- or ligament-forming cells, stimulate growth of tendon- or ligament-forming cells, induce differential ion 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, caφal tunnel syndrome and other tendon or ligament defects. The compositions may also include an appropriate matrix and/or sequestering agent as a carrier as is well known in the art. The protein of the present invention may also be useful for proliferation of neural cells and for regeneration of nerve and brain tissue, i.e. for the treatment of central and peripheral nervous system diseases and neuropathies, as well as mechanical and traumatic disorders , which involve degeneration, death or trauma to neural cells or nerve tissue. More specifically, a protein may be used in the treatment of diseases of the peripheral nervous system, such as peripheral nerve injuries, peripheral neuropathy and localized neuropathies, and central nervous system diseases, such as Alzheimer's, 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 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
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) ahd vascular (including vascular endothelium) tissue, or for promoting the growth of cells comprising such tissues Part of the desired effects may be by inhibition or modulation of fibrotic scarring to allow normal tissue to regenerate A protein of the invention may also exhibit angiogenic activity
A protein of the present invention may also be useful for gut protection or regeneration and treatment of lung or liver fibrosis, reperfusion injury in various tissues, and conditions resulting from systemic cytokine damage
A protein of the present invention may also be useful for promoting or inhibiting differentiation of tissues described above from precursor tissues or cells, or for inhibiting the growth of tissues described above
The activity of a protein of the invention may, among other means, be measured by the following methods
Assays for tissue generation activity include, without limitation, those described in International Patent Publication No W095/ 16035 (bone, cartilage, tendon), International Patent Publication No WO95/05846 (nerve, neuronal), International Patent Publication No W091/07491 (skin, endothelium )
Assays for wound healing activity include, without limitation, those described in Winter, Epidermal Wound Healing, pps 71-1 12 (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)
Activin/Inhibm 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 activms and are characterized by their ability to stimulate the release of follicle stimulating hormone (FSH) Thus, a protein of the present mventioa alone or in heterodimers with a member of the inhibin α 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 homodimeror as a heterodimer with other protein subunits of the inhibin-β 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 dnset of fertility in sexually immature mammals, so as to increase the lifetime reproductive performance of domestic animals such as cows, sheep and pigs.
The activity of a protein of the invention may, among other means, be measured by the following methods:
Assays for activin/inhibin activity include, without limitation, those described in: Vale et al. , Endocrinology 91 :562-572, 1972; Ling et al.. Nature 321 :779-782, 1986; Vale et al., Nature 321 :776-779, 1986; Mason et al.. Nature 318:659-663, 1985; Forage et al., Proc. Natl. Acad. Sci. USA 83:3091-3095, 1986.
Chemotactic/Chemokinetic Activity
A protein of the present invention may have chemotactic or chemokinetic activity (e.g., act as a chemokine) for mammalian cells, including, for example, monocytes, fibroblasts, neutrophils, T-cells, mast cells, eosinophils, epithelial and/or endothelial cells. Chemotactic and chemokinetic proteins can be used to mobilize or attract a desired cell population to a desired site of action. Chemotactic or chemokinetic proteins provide particular advantages in treatment of wounds and other trauma to tissues, as well as in treatment of localized infections. For example, attraction of lymphocytes, monocytes or neutrophils to tumors or sites of infection may result in improved immune responses against the tumor or infecting agent.
A protein or peptide has chemotactic activity for a particular cell population if it can stimulate, directly or indirectly, the directed orientation or movement of such cell 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 asέay for cell chemotaxis.
The activity of a protein of the invention may, among other means, be measured 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 include, without limitation, those described in: Current Protocols in Immunology, Ed by J.E. Coligan, A.M. Kruisbeek, D.H. Marguiies, E.M. Shevach, W. Strober, Pub. Greene Publishing Associates and Wiley-Interscience (Chapter 6.12, Measurement of alpha and beta Chemokines 6.12.1-6.12.28; Taub et al. J. Clin. Invest. 95: 1370-1376, 1995; Lind et al. APMIS 103: 140-146, 1995; Muller et al Eur. J. Immunol. 25: 1744-1748; Gruber et al. J. of Immunol. 152:5860-5867, 1994; Johnston et al. J. of Immunol. 153: 1762-1768, 1994.
Hemostatic and Thrombolytic Activity
A protein of the invention may also exhibit hemostatic or thrombolytic activity. As a result, such a protein is expected to be useful in treatment of various coagulation disorders (including hereditary disorders, such as 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.
Receptor/Ligand Activity
A protein of the present invention may also demonstrate activity as receptors, receptor ligands or inhibitors or agonists of receptor/ligand interactions. Examples of such receptors and ligands include, without limitation, cytokine receptors and their ligands, receptor kinases and their ligands, receptor phosphatases and their ligands, 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 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. Marguiies, 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: 1 145-1 156, 1988; Rosenstein et al. , J. Exp. Med. 169: 149-160 1989; Stoltenborg et al., J. Immunol. Methods 175:59-68. 1994; Stitt et al.. Cell 80:661-670, 1995.
Anti-Inflammatory Activity
Proteins of the present invention may also exhibit anti-inflammatory activity. The anti-inflammatory activity may be achieved by providing a stimulus to cells involved in the inflammatory response, by inhibiting or promoting cell-cell interactions (such as, for example, cell adhesion), by inhibiting or promoting chemotaxis of cells involved in the inflammatory process, inhibiting or promoting cell extravasation, or by stimulating or suppressing production of other factors which more directly inhibit or promote an inflammatory response. Proteins exhibiting such activities can be used to treat inflammatory conditions including chronic or acute conditions), including without limitation inflammation associated with infection (such as septic shock, sepsis or systemic inflammatory response syndrome (SIRS)), ischemia-reperfusioninjury, 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.
Tumor Inhibition Activity
In addition to the activities described above for immunological treatment or prevention of tumors, a protein of the invention may exhibit other anti-tumor activities. A protein may inhibit tumor growth directly or indirectly (such as, for example, via 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 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 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 or body part size or shape (such as, for example, breast augmentation or diminution, change in bone form or shape); effecting biorhythms or caricadic cycles or rhythms; effecting the fertility of male or female subjects; effecting the metabolism, catabolism, anabolism, processing, utilization, storage or elimination of dietary fat, lipid, protein, carbohydrate, vitamins, minerals, cofactors or other nutritional factors or component(s); effecting behavioral characteristics, including, without limitation, appetite, libido, stress, cognition (including cognitive disorders), depression (including depressive disorders) and violent behaviors; providing analgesic effects or other pain reducing effects; promoting differentiation and growth of embryonic stem cells in lineages other than hematopoietic lineages; hormonal or endocrine activity; in the case of enzymes, correcting deficiencies of the enzyme and treating deficiency-related diseases; treatment of hypeφroliferative disorders (such as, for example, psoriasis); immunoglobulin-like activity (such as, for example, the ability to bind antigens or complement); and the ability to act as an antigen in a vaccine composition to raise an immune response against such protein or another material or entity which is cross-reactive with such protein.
ADMINISTRATION AND DOSING
A protein of the present invention (from whatever source derived, including without limitation from recombinant and non-recombinantsources) may be used in a pharmaceutical composition when combined with a pharmaceutically acceptable carrier. Such a composition may also contain (in addition to protein and a carrier) diluents, fillers, salts, buffers, stabilizers, solubilizers, and other materials well known in the art. The term "pharmaceutically acceptable" means a non-toxic material that does not interfere with the effectiveness of the biological activity of the active ingredient(s). The characteristics of the carrier will depend on the route of administration. The pharmaceutical composition of the invention may also contain cytokines, lymphokines, or other hematopoietic factors such as M-CSF, GM-CSF, TNF, IL-1 , IL-2, IL-3, IL-4, IL-5, IL-6, IL-7, IL-8, IL-9, IL-10, IL-11, IL-12, IL-13, IL-14, IL-15, IFN, TNFO, TNF1 , TNF2, G-CSF, Meg-CSF, thrombopoietin, stem cell factor, and erythropoietin. The pharmaceutical composition may further contain other agents which either enhance the activity of the protein or compliment its activity or use in treatment. Such additional factors and/or agents may be included in the pharmaceutical composition to produce a synergistic effect with protein of the invention, or to minimize side effects. Conversely, protein of the present invention may be included in formulations of the particular cytokine, lymphokine, other hematopoietic factor, thrombolytic or anti-thrombotic factor, or anti-inflammaory agent to minimize side effects of the cytokine, lymphokine, other hematopoietic factor, thrombolytic or anti-thrombotic factor, or anti-inflammatory agent.
A protein of the present invention may be active in multimers (e.g., heterodimers or homodimers) or complexes with itself or other proteins. As a result, pharmaceutical compositions of the invention may comprise a protein of the invention in such multimeric or complexed form.
The pharmaceutical composition of the invention may be in the form of a complex of the protein(s) 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 antigen(s) to T lymphocytes. The antigen components could also* be supplied as purified MHC-peptide complexes alone or with co-stimulatory molecules that can directly signal T cells. Alternatively antibodies able to bind surface immunolgobulin and other molecules on B cells as well as antibodies able to bind the TCR and other molecules on T cells can be combined with the pharmaceutical composition of the invention
The pharmaceutical composition of the invention may be in the form of a liposome in which protein of the present invention is combined, in addition to other pharmaceutically acceptable carriers, with amphipathic agents such as lipids which exist in aggregated form as micelles, insoluble monolayers, liquid crystals, or lamellar layers m aqueous solution Suitable lipids for liposomal formulation include, without limitation, monoglycerides, diglyceπdes, sulfatides, lysolecithin, phospholipids, sapomn, bile acids, and the like Preparation of such liposomal formulations is within the level of skill in the art, as disclosed, for example, m 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 incoφorated 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 sufficient to show a meaningful patient benefit, l e , treatment, healing, prevention or amelioration of the relevant medical condition, or an increase in rate of treatment, healing, prevention or amelioration of such conditions When applied to an individual active ingredient, administered alone, the term refers to that ingredient alone When applied to a combination, the term refers to combined amounts of the active ingredients that result in the therapeutic effect, whether administered in combination, serially or simultaneously
In practicing the method of treatment or use of the present invention, a therapeutically effective amount of protein of the present invention is administered to a mammal having a condition to be treated Protein of the present invention may be administered in accordance with the method of the invention either alone or in combination with other therapies such as treatments employing cytokines, lymphokines or other hematopoietic factors When co-administered with one or more cytokines, lymphokines or other hematopoietic factors, protein of the present invention may be administered either simultaneously with the cytokme(s), lymphokme(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 m combination with cytokme(s), lymphokιne(s), other hematopoietic factor(s), thrombolytic or anti-thrombotic factors
Administration of protein of the present invention used in the pharmaceutical composition or to practice the method of the present invention can be carried out in a variety of conventional ways, such as oral ingestion, inhalation, topical application or cutaneous, subcutaneous, intraperitoneal, parenteral or intravenous injection Intravenous administration to the patient is preferred
When a therapeutically effective amount of protein of the present invention is administered orally, protein of the present invention will be in the form of a tablet, capsule, powder, solution or elixir When administered in tablet form, the pharmaceutical cOmposition of the invention may additionally contain a solid carrier such as a gelatin or an adjuvant The tablet, capsule, and powder contain from about 5 to 95% protein of the present invention, and preferably from about 25 to 90% protein of the present invention 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 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 effective amount of protein of the present invention is administered by intravenous, cutaneous or subcutaneous injection, protein of the present invention will be in the form of a pyrogen-free, parenterally acceptable aqueous solution The preparation of such parenterally acceptable protein solutions, having due regard to pH, isotonicity, stability, and the like, is within the skill in the art A preferred pharmaceutical composition for intravenous, cutaneous, or subcutaneous injection should contain, in addition to protein of the present invention, an isotonic vehicle such as Sodium Chloride Injection, Ringer's Injection, Dextrose Injection, Dextrose and Sodium Chloride Injection,
Lactated Ringer's Injection, or other vehicle as known in the art The pharmaceutical composition of the present invention may also contain stabilizers, preservatives, buffers, antioxidants, or other additives known to those of skill in the art
The amount of protein of the present invention in the pharmaceutical composition of the present invention will depend upon the nature and severity of the condition being treated, and on the nature of prior treatments which the patient has undergone Ultimately, the attending physician will decide the amount of protein of the present invention with which to treat each individual patient Initially, the attending physician will administer low doses of protein of the present invention and observe the patient's response Larger doses of protein of the present invention may be administered until the optimal therapeutic effect is obtained for the patient, and at that point the dosage is not increased further It is contemplated that the various pharmaceutical compositions used to practice the method of the present invention should contain about 0 01 μg to about 100 mg (preferably about 0 lμg to about 10 mg, more preferably about 0 1 μg to about 1 mg) of protein of the present invention per kg body weight
The duration of intravenous therapy using the pharmaceutical composition of the present invention will vary, depending on the severity of the disease being treated and the condition and potential idiosyncratic response of each individual patient It is contemplated that the duration of each application of the protein of the present invention will be in the range of 12 to 24 hours of continuous intravenous administration Ultimately the attending physician will decide on the appropriate duration of intravenous therapy using the pharmaceutical composition of the present invention
Protein of the invention may also be used to immunize animals to obtain 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 £5_, 2149-2154 (1963), J L Krstenansky, et al , FEBS Lett 211, 10 (1987) Monoclonal antibodies binding to the protein of the invention may be useful diagnostic agents for the immunodetection of the protein Neutralizing monoclonal antibodies binding to the protein may also be useful therapeutics for both conditions associated with the protein and also in the treatment of some forms of cancer where abnormal expression of the protein is involved In the case of cancerous cells or Ieukemic cells, neutralizing monoclonal antibodies against the protein may be useful in detecting and preventing the metastatic spread of the cancerous cells, which may be mediated by the protein
For compositions of the present invention which are useful for bone, cartilage, tendon or ligament regeneration, the therapeutic method includes administering 'the composition topically, systematically, or locally as an implant or device When administered, the therapeutic composition for use in this invention is, of course, in a pyrogen-free, physiologically acceptable form Further, the composition may desirably be encapsulated or injected in a viscous form for delivery to the site of bone, cartilage or tissue damage Topical administration may be suitable for wound healing and tissue repair Therapeutically useful agents other than a protein of the invention which may also optionally be included m 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 df bone and/or cartilage damage, providing a structure for the developing bone and cartilage and optimally capable of being resorbed into the body Such matrices may be formed of materials presently in use for other implanted medical applications The choice of matrix material is based on biocompatibility, biodegradability, mechanical properties, cosmetic appearance and interface properties The particular application of the compositions will define the appropriate formulation Potential matrices for the compositions may be biodegradable and chemically defined calcium sulfate, tπcalciumphosphate, hydroxyapatite, polylactic acid, polygiycohcacid and polyanhydπdes. Other potential materials are biodegradable and biologically well-defined, such as bone or dermal collagen Further matrices are comprised of pure proteins or extracellular matrix components Other potential matπces 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 tπcalciumphosphate The bioceramics may be altered in composition, such as in calcium-aluminate-phosphate and processing to alter pore size, particle size, particle shape, and biodegradability
Presently preferred is a 50 50 (mole weight) copolymer of lactic acid and glycolic acid in the form of porous particles having diameters ranging from 150 to 800 microns In some applications, it will be useful to utilize a sequestering agent, such as carboxymethyl cellulose or autologous blood clot, to prevent the protein compositions from disassociating from the matrix
A preferred family of sequestering agents is cellulosic materials such as alkylcelluloses(mcludinghydroxyalkylcelluloses), including methylcellulose, ethylcellulose, hydroxyethylcellulose, hydroxypropylcellulose, hydroxypropyl-methylcellulose, &nd 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 poly(vmyl alcohol) The amount of sequestering agent useful herein is 0 5-20 wt%, preferably 1-10 wt% based on total formulation weight, which represents the amount necessary to prevent desorbtion of the protein from the polymer matrix and to provide appropriate handling of the composition, yet not so much that the progenitor cells are prevented from infiltrating the matrix, thereby providing the protein the opportunity to assist the osteogenic activity of the progenitor cells.
In further compositions, proteins of the invention may be combined with other agents beneficial to the treatment of the bone and/or cartilage defect, wound, or tissue in question. These agents include various growth factors such as epidermal growth factor (EGF), platelet derived growth factor (PDGF), transforming growth factors (TGF-α and TGF-β), and insulin-like growth factor (IGF).
The therapeutic compositions are also presently valuable for veterinary applications. Particularly domestic animals and thoroughbred horses, in addition to humans, are desired patients for such treatment with proteins of the present invention.
The dosage regimen of a protein-containing pharmaceutical composition to be used in tissue regeneration will be determined by the attending physician considering various factors which modify the action of the proteins, e.g., amount of tissue weight desired to be formed, the site of damage, the condition of the damaged tissue, the size of a wound, type of damaged tissue (e.g., bone), the patient 's age, sex, and diet, the severity of any infection, time of administration and other clinical factors. The dosage may vary with the type of matrix used in the reconstitution and with inclusion of other proteins in the pharmaceutical composition. For example, the addition of other known growth factors, such as IGF I (insulin like growth factor I), to the final composition, may also effect the dosage. Progress can be monitored by periodic assessment of tissue/bone growth and/or repair, for example, X-rays, histomoφhometric 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 puφoses. Patent and literature references cited herein are incoφorated by reference as if fully set forth.
SEQUENCE LISTING
(1) GENERAL INFORMATION
(1) APPLICANT Jacobs, Kenneth McCoy, John LaVallie, Edward Racie, Lisa
Merberg, David Treacy, Maurice Evans, Cheryl
(11) TITLE OF INVENTION SECRETED PROTEINS
(ill) NUMBER OF SEQUENCES 59
(iv) CORRESPONDENCE ADDRESS
(A) ADDRESSEE Genetics Institute, Inc
(B) STREET 87 CambridgePark Drive
(C) CITY Cambridge
(D) STATE MA
(E) COUNTRY USA
(F) ZIP 02140
(v) COMPUTER READABLE FORM
(A) MEDIUM TYPE Floppy disk
(B) COMPUTER IBM PC compatible
(C) OPERATING SYSTEM PC DOS/MS DOS
(D) SOFTWARE Patentln Release #1 0 Version #1 30
(vi) CURRENT APPLICATION DATA
(A) APPLICATION NUMBER
(B) FILING DATE
(C) CLASSIFICATION
(vm) ATTORNEY/AGENT INFORMATION
(A) NAME Brown, Scott A
(B) REGISTRATION NUMBER 32,724
(C) REFERENCE/DOCKET NUMBER GI6001
(ix) TELECOMMUNICATION INFORMATION
(A) TELEPHONE (617) 498 8224
(B) TELEFAX (617) 876-5851
(2) INFORMATION FOR SEQ ID NO 1
(l) SEQUENCE CHARACTERISTICS
(A) LENGTH 505 base pairs
(B) TYPE nucleic acid
(C) STRANDEDNESS double
(D) TOPOLOGY linear
In) MOLECULE TYPE cDNA
(xi) SEQUENCE DESCRIPTION SEQ ID NO 1 GAATTCGGCA CGAGGGCGCG GGGTCCGYWA TGGCGSCGGC AGCCGAAGGC GTACTGGCGA 60
CCCGGAGTGA TGAGCCCGCC CGAGACGATG CCSCCGTGGA GACAGCTGAG GAARCAAAGG 120
AGCCTGCTGA AAGCTGACAT CACTGAGCTC TGCCGGGACA TGTTCTCCAA AATGGCCACT 180
TACCTGACTG GGGAACTGAC GGCCACCAGT GAAGACTATA AGCTCCTGGA AAATATGAAT 240
AAACTCACCA GCTTGAAGTA TYTTGAAATG AAAGATATTG CTATAAACAT TAGTAGGAAC 300
TTAAAGGACT TAAACCAGAA ATATGCTGGA CTGCAGCCTT ATYTGGATTC AGATTCAATG 360
TTCATTGGAA GAGCAGGTAG CAGCTTTTTG AGCAGGCAGC TTACAAGTTG GRTGCMTWTT 420
TCAAAAAAAN TGGAANCCCA AGTACAAGAA GNTGGAGAAG CGATGAGAAA ATTATTTTTA 480
TGGGACAGAG ττττττττττ TTAAT 505 (2) INFORMATION FOR SEQ ID NO 2
(l) SEQUENCE CHARACTERISTICS
(A) LENGTH 145 ammo acids
(B) TYPE ammo acid
(C) STRANDEDNESS
(D) TOPOLOGY linear
111) MOLECULE TYPE protein
(xi) SEQUENCE DESCRIPTION SEQ ID NO: 2
Met Ser Pro Pro Glu Thr Met Pro Pro Trp Arg Gin Leu Arg Lys Gin 1 5 10 15
Arg Ser Leu Leu Lys Ala Asp lie Thr Glu Leu Cys Arg Asp Met Phe 20 25 30
Ser Lys Met Ala Thr Tyr Leu Thr Gly Glu Leu Thr Ala Thr Ser Glu 35 40 45
Asp Tyr Lys Leu Leu Glu Asn Met Asn Lys Leu Thr Ser Leu Lys Tyr 50 55 60
Xaa Glu Met Lys Asp lie Ala He Asn He Ser Arg Asn Leu Lys Asp 65 70 75 80
Leu Asn Gin Lys Tyr Ala Gly Leu Gin Pro Tyr Leu Asp Ser Asp Ser 85 90 95
Met Phe He Gly Arg Ala Gly Ser Ser Phe Leu Ser Arg Gin Leu Thr 100 105 110
Ser Trp Xaa Xaa Xaa Ser Lys Lys Xaa Glu Xaa Gin Val Gin Glu Xaa
115 120 125
Gly Glu Ala Met Arg Lys Leu Phe Leu Trp Asp Arg Val Phe Phe Phe 130 135 140
Xaa
145 (2) INFORMATION FOR SEQ ID NO 3
(l) SEQUENCE CHARACTERISTICS
(A) LENGTH 315 base pairs
(B) TYPE nucleic acid
(C) STRANDEDNESS double
(D) TOPOLOGY linear
In) MOLECULE TYPE cDNA
(xi) SEQUENCE DESCRIPTION SEQ ID NO 3
GCAGCTTATC ACCTTGTGAA TGTCGGTAAC TTACTTTTCC ATAATATTGC AAATAACATA 60
AAATNTTAAA ATAATTCCAA GCTGAGTTTT CTAGATTGAG CAGAAATGGT GAAAGGAGTA 120
TTGATAACTT GGCGTATGTG ATGGGCCCCT CTTGTTTATT TTNTATGTGA GTCACATTGA 180
CATGCGATCA GTTTGGGGAA ATGTGATGAA AACAAAGACT AGATGGGTAT GTGTGTTTAT 240
GTGTTGGGTA GGGAGGTGAC GATTGCCANT CATANAATAA AGGATTTTAT AAAATACCAA 300
AAAAAAAAAA AAAAA 315 (2) INFORMATION FOR SEQ ID NO 4
(l) SEQUENCE CHARACTERISTICS
(A) LENGTH 867 base pairs
(B) TYPE nucleic acid
(C) STRANDEDNESS double
(D) TOPOLOGY linear
111) MOLECULE TYPE cDNA
(XI ) SEQUENCE DESCRIPTION SEQ ID NO 4
GTCAGTNTGA GTNAGAAAAT NGAATCCATC ATCTAACAGA GANTTTCATC CAGAAACAGR 60
CCATGYTGGA GAGTCTCAGC ACAGAAAAGA ACTNCCTGGT CTTTCAACTG GAGCGCCTCG 120
AACAGCAGAT GAACTCCGCC TCTGGAAGTA GTAGTAATGG GTCTTCGATT AATATGTCTG 180
GAATTGACAA TGGTGAAGGC ACTCGTCTGC GAATGTTCCT GTTCTTTTTA ATGACACAGA 240
AACTΛATCTG GCAGGAATGT ACGGAAAAGT TCGCAAAGCT GCTAGTTCAA TTGATCAGTT 300
TAGTATTCGC CTGGGNAATT TTTCTCCGAA GATACCCCAT AGCGCGAGTT TTTGTAATTA 360
TATATATGGC TTTGCTTCAC CTCTGGGTNA TGATTGTTCT GTTGACTTAC ACACCAGAAA 420
TGCACCACGA CCAACCATAT GGCAAATGAA CCAΛGCCCAG TTGTTGCAGT GATTGGTTGT 480
CTTTTTYTAG ACTTGGGATY TGCAAGAAGG CCAATTGCCT AAAATTTTTG AGAACAGTGC 540
ACAAGATTAT TTTATCANTA CAAGNTTTTA AANTTTTTAA GTTATTGNAN AAGTATTTTA 600 CCTAAATTTT CCAATTTCCT TTAAATGGTA AGAGTTTTTA AAACAGACAA TAATTTAACA 660
AGNTCAGNTT TGCTTTATTT GAGTTTAGTG GTCCTAATAT ATATGTAGAG AAAGATGGTG 720
GGGTTGTTCA CCTCTGTACA GGACCTTTTG TATGTTAGGN GACATTGATT ATGGGTTATA 780
ATCAGGGAAA CTAATTGTAT TTAGTGACAA AAATAAAAAG NTTTTTTTTT TATNAAAAAA 840
AAAAAAAAAA AAAAAAAAAA AATTATT 867 (2) INFORMATION FOR SEQ ID NO: 5 :
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 212 ammo acids
(B) TYPE: amino acid
(C) STRANDEDNESS:
(D) TOPOLOGY: linear
(ii) MOLECULE TYPE: protein
(Xl) SEQUENCE DESCRIPTION: SEQ ID NO: 5 :
Met Thr Gin Lys Leu He Trp Gin Glu Cys Thr Glu Lys Phe Ala Lys 1 5 10 15
Leu Leu Val Gin Leu He Ser Leu Val Phe Ala Trp Xaa He Phe Leu 20 25 30
Arg Arg Tyr Pro He Ala Arg Val Phe Val He He Tyr Met Ala Leu 35 40 45
Leu His Leu Trp Val Met He Val Leu Leu Thr Tyr Thr Pro Glu Met 50 55 60
His His Asp Gin Pro Tyr Gly Lys Xaa Thr Lys Pro Ser Cys Cys Ser 65 70 75 80
Asp Trp Leu Ser Phe Xaa Arg Leu Gly He Cys Lys Lys Ala Asn Cys 85 90 95
Leu Lys Phe Leu Arg Thr Val His Lys He He Leu Ser Xaa Gin Xaa 100 105 110
Phe Lys Xaa Phe Lys Leu Leu Xaa Lys Tyr Phe Thr Xaa He Phe Gin 115 120 125
Phe Pro Leu Asn Gly Lys Ser Phe Xaa Asn Arg Gin Xaa Phe Asn Lys 130 135 140
Xaa Xaa Phe Ala Leu Phe Glu Phe Ser Gly Pro Asn He Tyr Val Glu 145 150 155 160
Lys Asp Gly Gly Val Val His Leu Cys Thr Gly Pro Phe Val Cys Xaa 165 170 175
Xaa Thr Leu He Met Gly Tyr Asn Gin Gly Asn
(2) INFORMATION FOR SEQ ID NO: 6 : (l) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 491 base pairs
(B) TYPE: nucleic acid
(C) STRANDEDNESS: double
(D) TOPOLOGY: linear
111) MOLECULE TYPE: cDNA
(xil SEQUENCE DESCRIPTION: SEQ ID NO: 6 :
CAGGATATTA GAAATGGCTA CTCCCCAGTC AATTTTCATC TTTGCAATCT GCATTTTAAT 60
GATAACAGAA TTAATTCTGG CCTCAAAAAG CTACTATGAT ATCTTAGGTG TGCCAAAATC 120
GGCATCAGAG CGCCAAATCA AGAAGGCCTT TCACAAGTTG GCCATGAAGT ACCACCCTGA 180
CAAAAATAAG AGCCCGGATG CTGAAGCAAA ATTCAGAGAG ATTGCAGAAG CATATGAAAC 240
ACTCTCAGAT GCTAATAGNA CGAAAAGAGT ATGATACACT TGGACACAGT GCTTTTACTA 300
GTGGGTAAAG GGACAARGRR GTAGTTGGRA GTTCTTTTGA GYRNKCNKTT MNYTTYAAYT 360
TTSATGACTT ATTTAAAGAC TTTGGCTTTT TTGGTYNARR CYAAAACAYT GGAKCYAANA 420
AYKTTTTGRR RWYCAWWYCC NNACACCCNN NWKGGTKSYC CAGGNGGCGT TTTTTTGNAA 480
TTCCTTTTCC C 491 (2) INFORMATION FOR SEQ ID NO : 7 :
(l) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 159 amino acids
(B) TYPE: ammo acid
(C) STRANDEDNESS:
(D) TOPOLOGY: linear
(ll) MOLECULE TYPE, protem
(XI) SEQUENCE DESCRIPTION: SEQ ID NO: 7
Met Ala Thr Pro Gin Ser He Phe He Phe Ala He Cys He Leu Met 1 5 10 15
He Thr Glu Leu He Leu Ala Ser Lys Ser Tyr Tyr Asp He Leu Gly 20 25 30
Val Pro Lys Ser Ala Ser Glu Arg Gin He Lys Lys Ala Phe His Lys 35 40 45
Leu Ala Met Lys Tyr His Pro Asp Lys Asn Lys Ser Pro Asp Ala Glu 50 55 60
Ala Lys Phe Arg Glu He Ala Glu Ala Tyr Glu Thr Leu Ser Asp Ala 65 70 75 80
Asn Xaa Thr Lys Arg Val Xaa Tyr Thr Trp Thr Gin Cys Phe Tyr Xaa 85 90 95
Trp Val Lys Gly Gin Xaa Xaa Ser Trp Xaa Phe Phe Xaa Xaa Xaa Xaa 100 105 110
Xaa Xaa Xaa Xaa Xaa Xaa Leu He Xaa Arg Leu Trp Leu Phe Trp Xaa 115 120 125
Xaa Xaa Lys His Trp Xaa Xaa Xaa Xaa Phe Xaa Xaa Xaa Xaa Xaa Thr 130 135 140
Pr6 Xaa Xaa Val Xaa Gin Xaa Ala Phe Phe Xaa Asn Ser Phe Ser
145 150 155
(2) INFORMATION FOR SEQ ID NO 8-
(l) SEQUENCE CHARACTERISTICS
(A) LENGTH: 242 base pairs
(B) TYPE: nucleic acid
(C) STRANDEDNESS. double
(D) TOPOLOGY linear
In) MOLECULE TYPE cDNA
(xi) SEQUENCE DESCRIPTION SEQ ID NO• 8
CTTAATCTAG AGATTGACTG ANACCTCATT CTGTTNGTAA AACCAGCCAG TAATTTCTGT 60
GCAACCTTAC TATGTGCAAT ATTTTTAAAT CCTGAGAAAT GTGTGCTTTT GTTTTCGGAT 120
AGACTTATTT CTTTAGTTCT GCACTTTTCC ACATTATACT CCATATGAGT ATTAATCCTA 180
TGGATAACAT ATTAAAACAA GTGTCTCATA AAAAAAAAAA AAAAAAAATT NCCTGCGGCC 240 GC 242
(2) INFORMATION FOR SEQ ID NO 9
(l) SEQUENCE CHARACTERISTICS
(A) LENGTH. 607 base pairs
(B) TYPE: nucleic acid
(C) STRANDEDNESS: double
(D) TOPOLOGY: linear
(11) MOLECULE TYPE: cDNA
(xi) SEQUENCE DESCRIPTION. SEQ ID NO: 9-
GAATTCNCNG CCCTACAGCA CGGCCCTGCC CCAAGGACTT TTGNTGTCCT TGGCCAGTTT 60
CTGGTGCTAA AGAAAAGATN RAARACCTCT TCCGGGAATG GCTGAAAGAC ACTTGTGGCG 120
CCAACGCCAA GCAGTCCCGG GACTGCTTCG GATGCCTTCG AGAGTGGTGC GACGCCTTCT 180
KGTGATGCTC TCTGGGAARC TCTCAATCCC CAGCCCTCAT CCAGAGTTTG CAGCCGAGTA 240 GGGACTCNTC CCCTGTCHTT TACGAAGGAA AAGATTGCTA TTGTCGTACT CACNTCNGAC 300
GTANTCCGGG GTNTTTTGGG AGTTTTCTCC CCTAACCATT TCAACTTTTT TTGGATTHTC 360
GNTCTTGCAT GCCTCCCCCG TCCTTTTTCC CTTGCCAGTT CCCTGGTGAA CAGTTTACCA 420
GCTTTTCCTG AATGGATTNC CGGΞCCCCAT CCCTCACCCC CACCYTCAAT TTCAATTCCG 480
TTTTGATAMC ATTKGGYTCC TTTTTTTGGC AGAACAGTCA MTGTCCTTGT AAAGTTTTTT 540
AGATCAATAA AGTCAGTGGC TTTCAAAAAN GNAAAAAAAA AAAAAAAAAA AAAAAAAGGG 600 CGGCCGC 607
(2) INFORMATION FOR SEQ ID NO: 10: d) SEQUENCE CHARACTERISTICS
(A) LENGTH 202 amino acids
(B) TYPE- amino acid
(C) STRANDEDNESS-
(D) TOPOLOGY linear
In) MOLECULE TYPE protein
(xi) SEQUENCE DESCRIPTION- SEQ ID NO.10.
Glu Phe Xaa Ala Leu Gin His Gly Pro Ala Pro Arg Thr Phe Xaa Val 1 5 10 15
Leu Gly Gin Phe Leu Val Leu Lys Lys Arg Xaa Lys Thr Ser Ser Gly 20 25 30
Asn Gly Xaa Lys Thr Leu Val Ala Pro Thr Pro Ser Ser Pro Gly Thr 35 40 45
Ala Ser Asp Ala Phe Glu Ser Gly Ala Thr Pro Ser Xaa Asp Ala Leu 50 55 60
Trp Glu Xaa Leu Asn Pro Gin Pro Ser Ser Arg Val Cys Ser Arg Val 65 70 75 80
Gly Thr Xaa Pro Leu Ser Phe Thr Lys Glu Lys He Ala He Val Val 85 90 95
Leu Thr Ser Asp Val Xaa Arg Gly Xaa Leu Gly Val Phe Ser Pro Asn 100 105 110
His Phe Asn Phe Phe Trp He Xaa Xaa Leu Ala Cys Leu Pro Arg Pro 115 120 125
Phe Ser Leu Ala Ser Ser Leu Val Asn Ser Leu Pro Ala Phe Pro Glu 130 135 140
Trp He
(2) INFORMATION FOR SEQ ID NO: 11. ( 1 ) ΞEQUENCE CHARACTERISTICS . (A) LENGTH: 462 base pairε
(B) TYPE: nucleic acid
(C) STRANDEDNESS: double
(D) TOPOLOGY: linear
(li) MOLECULE TYPE: cDNA
(xi) SEQUENCE DESCRIPTION: SEQ ID NO: 11:
AGCTTGGAAR AARRATGAAA TTCCTTATCT TCGCATTTTT CGGTGGTGTT CACCTTTTAT 60
CCCTGTGCTC TGGGAAAGCT ATATGCAAGA ATGGCATCTC TAAGAGGACT TTTGAAGAAA 120
TAAAAGAAGA AATAGCCAGC TGTGGAGATG TTGCTAAAGC AATCATCAAC CTAGCTGTTT 180
ATGGTAAAGC CCAGAACAGA TCCTATGAGC GATTGGCACT TCTGGTTGAT ACTGTTGGAC 240
CCAGACTGAG TGGCTCCAAG AACCTΛGRAA AAAGCCATCC AAATTATGTA CCAAAACCTG 300
GCAGGCAAGA TGGGGCTGGG AGGAAAGTTC ACCTGGGGAG CCAGTGAGGA ATACCCCACT 360
GGGGAGGAGG GGGGAGAAGG ATNCAGCTGT TGATNGCTGG GAGCCCAAGG ATTTCATTAA 420
GGTTAGGCCN TCCTGGGGTC TTTTGGCCAG CCAGCNTTTG GG 462 (2) INFORMATION FOR SEQ ID NO: 12:
(l) SEQUENCE CHARACTERISTICS-
(A) LENGTH. 149 ammo acids
Figure imgf000083_0001
(C) STRANDEDNESS:
(D) TOPOLOGY: linear
In) MOLECULE TYPE: protein
(xi) SEQUENCE DESCRIPTION- SEQ ID NO: 12:
Met Lys Phe Leu He Phe Ala Phe Phe Gly Gly Val His Leu Leu Ser 1 5 10 15
Leu Cys Ser Gly Lys Ala He Cys Lys Asn Gly He Ser Lys Arg Thr 20 25 30
Phe Glu Glu He Lys Glu Glu He Ala Ser Cys Gly Asp Val Ala Lys 35 40 45
Ala He He Asn Leu Ala Val Tyr Gly Lys Ala Gin Asn Arg Ser Tyr 50 55 60
Glu Arg Leu Ala Leu Leu Val Asp Thr Val Gly Pro Arg Leu Ser Gly 65 70 75 80
Ser Lys Asn Leu Xaa Lys Ser His Pro Asn Tyr Val Pro Lys Pro Gly 85 90 95
Arg Gin Asp Gly Ala Gly Arg Lys Val His Leu Gly Ser Gin Xaa Gly 100 105 110
He Pro His Trp Gly Gly Gly Gly Arg Arg Xaa Gin Leu Leu Xaa Ala 115 120 125
Gly Ser Pro Arg He Ser Leu Arg Leu Gly Xaa Pro Gly Val Phe Trp 130 135 140
Pro Ala Ser Xaa Trp 145
(2) INFORMATION FOR SEQ ID NO 13 d) SEQUENCE CHARACTERISTICS
(A) LENGTH 360 base pairs
(B) TYPE nucleic acid
(C) STRANDEDNESS double
(D) TOPOLOGY linear
(π) MOLECULE TYPE cDNA
(xi) SEQUENCE DESCRIPTION SEQ ID NO 13
AGAAACAGTA AGAAAGAAAC GTTTTCATGN TTCTGGCCAG GAATCCTGGG TCTGCAACTT 60
NGGAAAACTC NTCTTCACAT AACAATTTCA TCCAATTCAT NTTCAAAGCA CAACTNTATT 120
TCATGCTTTC TGNNANNATA TTTCTTGATA CTTTCCAAAT TCTCTGATTC TAGAAAAAGG 180
AATCATTNTC CCCTCCCTCC CACCACATAG AATCAACATA TGGTAGGGAT TACAGTGGGG 240
GCATTTCTTT ATATCACCTC TTAAAAACAT TGTTTCCACT TTAAAAGTAA ACACTTAATA 300
AATTTTTGGA AGATCTCTGA AAAAAAAAAA AAAAAAAAAA AAAAATTNCC TGCGGCCGCA 360 (2) INFORMATION FOR SEQ ID NO 14
(l) SEQUENCI CHARACTERISTICS
(A) LENGTH 519 base pairs
(B) TYPE nucleic acid
(C) STRANDEDNESS double
(D) TOPOLOGY linear
(li) MOLECULE TYPE cDNA
(xi) SEQUENCE DESCRIPTION SEQ ID NO 14
AAGCTTGGCA CGAGGGGACC CGGCGCTCTC CCCGTGTCCT CTCCACGACT CGCTCGGCCC 60
CTCTGGAATA AAACACCCGC GAGCCCCGAG GGCCCAGAGG AGGCCGACGT GCCCGAGCTC 120
CTCCGGGGGT CCCGCCCGCG AGCTTTCTTC TCGCCTTCGC ATCTCCTCCT CGCGCGTCTT 180
GGACATGCCA GGAATAAAAA GGATACTCAC TGTTACCATT CTGGCTCTCT GTCTTCCAAG 240
CCCTGGGAAT GCACAGGCAC AGTGCACGAA TGGCTTTGAC CTGGATCGCC AGTCAGGACA 300 GTGTTTAGAT ATTGATGAAT GCCGAACCAT CCCCGAGGCC TGCCGAGGAG ACATGATGTG 360
TGTTAACCAA AATGGCGGGT ATTTATGCAT TCCCCGGACA AACCCTGTGT ATCGAGGGCC 420
NTACTCGAAC CCCTACTCGA CCCCTTAYTC AGGTCCGTAA CCCAGCAGYT GGCCCCACCA 480
YTTTACAGYT CCAAAYTTTC CAAKGTTTTT CAGGGTTTT 519 (2) INFORMATION FOR SEQ ID NO: 15:
(i) SEQUENCE CHARACTERISTICS-
(A) LENGTH. Ill amino acids
(B) TYPE: amino acid
(C) STRANDEDNESS:
(D) TOPOLOGY: linear
(n) MOLECULE TYPE: protein
(xi) SEQUENCE DESCRIPTION: SEQ ID NO: 15
Met Pro Gly He Lys Arg He Leu Thr Val Thr He Leu Ala Leu Cys
1 5 10 15
Leu Pro Ser Pro Gly Asn Ala Gin Ala Gin Cys Thr Asn Gly Phe Asp 20 25 30
Leu Asp Arg Gin Ser Gly Gin Cys Leu Asp He Asp Glu Cys Arg Thr 35 40 45
He Pro Glu Ala Cys Arg Gly Asp Met Met Cys Val Asn Gin Asn Gly 50 55 60
Gly Tyr Leu Cys He Pro Arg Thr Asn Pro Val Tyr Arg Gly Pro Tyr 65 70 75 80
Ser Asn Pro Tyr Ser Thr Pro Tyr Ser Gly Pro Xaa Pro Ser Ser Trp 85 90 95
Pro His His Phe Thr Xaa Pro Asn Phe Pro Xaa Phe Phe Arg Val 100 105 110
(2) INFORMATION FOR SEQ ID NO: 16.
(l) SEQUENCE CHARACTERISTICS: (A) LENGTH: 54 base pairs
(l) SEQUENCE CHARACTERISTICS
(A) LENGTH 536 base pairs
(B) TYPE: nucleic acid
(C) STRANDEDNESS: double
(D) TOPOLOGY, linear
In) MOLECULE TYPE. cDNA
(xi? SEQUENCE DESCRIPTION: SEQ ID NO.17
GTGGAATTTG TGGGTAGTGT GATNTTTGTT TGTATCCTTT TAAGTACTGT TGATCAGTTG 60
NGACACTTAC TGGTTAAACT TACGTTGCTA AAGATTTCTC TATAATAAGC CACACATTAT 120
ATTTAGACTA TATTAAGGGA CCTTGGTTTT CTTCTAGATA GCAGCTGTCC CAAAGAAAAT 180
ATTTCTTCTT TGTCTGTTAA GATTTAGCTA TTATCTGCCA GTTGTTAAGA GGTTTTGGTT 240
CCAAACTCAA CCAGCAATGT TGAGAGCTGA ΛCTTAAGATA GCTGTTGTAC TTTTTGCTTT 300
CCATCTGTTA CTGTCCTTCA TTCTTGGCTC CCTACTATCT ATAAACAGCT GCTGTGAAGG 360
AAGGAAAAGT TGAATAAGGA GTTGGGCTTA AATTTTAAAA AAGGAAAAAR GAAAATTGAG 420
GTTTTAGGRT TTTCATGGGT AACAAGCTCT GGGTATTARG CTAAGGCTGG GCAAGTTTCA 480
GGWTACTAAA ATATTATTTG ATCATATCTT GGΛTCCNTAT YYTGRRAAAT TTAAAA 536 (2) INFORMATION FOR SEQ ID NO.18:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH. 93 amino acids
(B) TYPE: arrmo acid
(C) STRANDEDNESS:
(D) TOPOLOGY linear
(n) MOLECULE TYPE: protein
(xi) SEQUENCE DESCRIPTION: SEQ ID NO:18:
Met Leu Arg Ala Glu Leu Lys He Ala Val Val Leu Phe Ala Phe His 1 5 10 15
Leu Leu Leu Ser Phe He Leu Gly Ser Leu Leu Ser He Asn Ser Cys 20 25 30
Cys Glu Gly Arg Lys Ser Xaa He Arg Ser Trp Ala Xaa He Leu Lys 35 40 45
Lys Glu Lys Xaa Lys Leu Arg Phe Xaa Xaa Phe His Gly Xaa Gin Ala 50 55 SO
Leu Gly He Xaa Leu Arg Leu Gly Lys Phe Gin Xaa Thr Lys He Leu 65 70 75 80
Phe Asp His He Leu Asp Pro Tyr Xaa Xaa Lys Phe Lys (2) INFORMATION FOR SEQ ID NO 19
(l) SEQUENCE CHARACTERISTICS
(A) LENGTH 397 base pairs IB) TYPE nucleic acid
(C) STRANDEDNESS double
(D) TOPOLOGY linear
In) MOLECULE TYPE cDNA
(xi) SEQUENCE DESCRIPTION SEQ ID NO 19
AAGGTAAATT AGAAATAAGT ATGAATATTA ATAAAATAGC ATTTATCTTA TTTCTCTATT 60
TTATGTTGTG ACTTAACCTA ATTTTATTTT TTTAACATTT TCTTATTTCT TATAATATGA 120
ATGCTGATAT TTAAAGGTAG ATCTATGTGG TATTCTTTGT GTTTCTNAAT TGTATAGCTC 180
TTAAGATTAT TTGTGATCTG GATTTATGTA TTTGTTAGAT ACATACGAAT TGTTAAAATG 240
GAATGCAAGT TTTTCAAAAG CCCAGGTCTA AATGTAATGG TTGGTTTATT GTTCTATAAC 300
CCCAGCCCΛT CATTTTCTGT GTAAATCATA AACAATAAAC AGAATATACT CGGTGGTCAT 360
TTCTAAAAAA AAAAAAAAAA AAATTNCCTG CGGCCGC 397
(2) INFORMATION FOR SEQ ID NO 20
(l) SEQUENCE CHARACTERISTICS
(A) LENGTH 476 base pairs IB) TYPE nucleic acid
(C) STRANDEDNESS double
(D) TOPOLOGY linear
(n) MOLECULE TYPE cDNA
(xi) SEQUENCE DESCRIPTION SEQ ID NO 20
GAATTCGGCA CNAGCAGTGA AGCGCAGTGA CAGCAGTGGG AACCGGAATA TCCAAAGAGT 60
GGTTTGAAGG AGAAAGAAGC ATTGTGGCTT TATATCCTCT GGGCCTGGGT TTCCTGAAGT 120
CACCACACAT AGAGGAGAGA GAAAATGGCT GAGTTAAAGT ACATTTCTGG ATTTGGGAAT 180
GAGTGTTCTT CAGAGGATCC TCGCTGCCCA GGTTCCCTGC CAGAAGGACA GAATAATCCT 240
CAGGTCTGCC CCTACAATCT CTATGCTGAG CAGCTCTCAG GATCGGCTTT CACTTGTCCA 300
CGGAGCACCA ATAANGAGAA GCTGGCTGTA TAGGATTCTA CCTTCAGTTT YTCACAAGCC 360
CTTTGGAATC CATTTGACGA NGGCCAYGTT CACTCACAAC TGGGGNATGG AAGTTGATCC 420
TGATCCTAAC CAGNTTAGAT GGNAAACCAT TTTTGAGGTT TCCAAAAGGC ATNTTC 476
85 (2) INFORMATION FOR SEQ ID NO: 21:
(l) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 99 ammo acids
(B) TYPE: ammo acid
(C) STRANDEDNESS:
(D) TOPOLOGY, linear
(a) MOLECULE TYPE: protein
(xi) SEQUENCE DESCRIPTION: SEQ ID NO:21.
Met Ser Val Leu Gin Arg He Leu Ala Ala Gin Val Pro Cys Gin Lys 1 5 10 15
Asp Arg He He Leu Arg Ser Ala Pro Thr He Ser Met Leu Ser Ser 20 25 30
Ser Gin Asp Arg Leu Ser Leu Val His Gly Ala Pro He Xaa Arg Ser 35 40 45
Trp Leu Tyr Arg He Leu Pro Ser Val Xaa His Lys Pro Phe Gly He 50 55 60
His Leu Thr Xaa Ala Xaa Phe Thr His Asn Trp Gly Met Glu Val Asp 65 70 75 80
Pro Asp Pro Asn Gin Xaa Arg Trp Xaa Thr He Phe Glu Val Ser Lys 85 90 95
Arg His Xaa
(2) INFORMATION FOR SEQ ID NO: 22:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 49 base pairs
(B) TYPE: nucleic acid
(C) STRANDEDNESS: double
(D) TOPOLOGY, linear
111) MOLECULE TYPE: cDNA
(xi) SEQUENCE DESCRIPTION: SEQ ID NO: 22: GGGGAAAAAA AAAAAAAAAA AAAAAAAAAA AAAAAAAAAA AAACTCGAG 49
(2) INFORMATION FOR SEQ ID NO:23:
(l) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 612 base pairs
(B) TYPE: nucleic acid
(C) STRANDEDNESS: double
(D) TOPOLOGY: linear
(li) MOLECULE TYPE: cDNA (xi) SEQUENCE DESCRIPTION SEQ ID NO 23
AAGCTTGGCA CGAGGCAGGG AAGGTCCTGA CCCCANCGAG CACTTCTGAC AATGAGACCA 60
GAGACTCCWC AATTATTGAT CCAGGAACTG AGCAAGATCT TCCTTCCCCT GAAAATAGTT 120
CTGTTAAAGA ATACCGAATG GAAGTTCCAT CTTCGTTTTC AGAAGACATG TCAAATATCA 180
GGTCACAGCA TGCAGAAGAA CAGTCCAACA ATGGTAGATA TGACGATTGT AAAGAATTTA 240
AAGACCTCCA CTGTTCCAAG GATTMTACCC TAGCCGAGGA AGAATCTGAG TTCCCTTCTA 300
CTTCTATCTC TGCAGTTCTG TCTGACTTAG CTGACTTGAG AAGCTGTGAT GGCCAAGCTT 360
TGCCCTCCCA GGGACCCTGA GGTTGCTTTA TCTCTCAGTT GTGGCCATTC CAGAGGACTC 420
TTTAGTCATA TGCAGCAACA TGACATTTTA GGATACCCTG TGTTAGGGAC CATTGAATCT 480
ACAATCCATG TTCGTTCACA AGGGATATCT GGGCAAAGGG AAACCAAGCT GCTTCTTTGA 540
ACATTAGGGN GTTAGGCATT GTCTTACTTT TTAAAGTCCC TCACCCCCAA CCCCCATGCT 600
GTTTTGTATA AG 612 (2) INFORMATION FOR SEQ ID NO 24
(l) SEQUENCE CHARACTERISTICS
(A) LENGTH 131 amino acids
(B) TYPE amino acid
(C) STRANDEDNESS
(D) TOPOLOGY linear
In) MOLECULE TYPE protem
(xi) SEQUENCE DESCRIPTION SEQ ID NO 24
Met Thr He Val Lys Asn Leu Lys Thr Ser Thr Val Pro Arg He Xaa 1 5 10 15
Pro Xaa Pro Arg Lys Asn Leu Ser Ser Leu Leu Leu Leu Ser Leu Gin 20 25 30
Phe Cys Leu Thr Xaa Leu Thr Xaa Glu Ala Val Met Ala Lys Leu Cys 35 40 45
Pro Pro Arg Asp Pro Glu Val Ala Leu Ser Leu Ser Cys Gly His Ser 50 55 60
Arg Gly Leu Phe Ser His Met Gin Gin His Asp He Leu Gly Tyr Pro 65 70 75 80
Val Leu Gly Thr He Glu Ser Thr He His Val Arg Ser Gin Gly He 85 90 95
Ser Gly Gin Arg Glu Thr Lys Leu Leu Leu Xaa Thr Leu Gly Xaa Xaa 100 105 110 Ala Leu Ser Tyr Phe Leu Lys Ser Leu Thr Pro Asn Pro His Ala Val 115 120 125
Leu Tyr Lys 130
(2) INFORMATION FOR SEQ ID NO 25
(l) SEQUENCE CHARACTERISTICS
(A) LENGTH 69 base pairs
(B) TYPE, nucleic acid
(C) STRANDEDNESS double
(D) TOPOLOGY linear
In) MOLECULE TYPE cDNA
(xi) SEQUENCE DESCRIPTION SEQ ID NO 25
TTTTTAAAAA AAAAAAAAAA AAAAAAAAAA AAAAAAAAAA AAAAAAAAAA AAATTNTCNC 60
TGCGGCCGC 69
(2) INFORMATION FOR SEQ ID NO 26 (l) SEQUENCE CHARACTERISTICS
(A) LENGTH 655 base pairs IB) TYPE nucleic acid
(C) STRANDEDNESS double
(D) TOPOLOGY linear
In) MOLECULE TYPE cDNA
(xi) SEQUENCE DESCRIPTION SEQ ID NO 26
CAATNATAAA ATGTCAGCTT TTAAGGNANN CCTGTGGAAT ATATTTTCCA GCAATAAAAA 60
GAGATCCAGG CAGATATTTA CATAGTTGTC CCTGAATCTG TGAAAAAATG GCTTCGACAG 120
CTAAAGAATG CTGGGAAAAT TCTTCTGTTA ATNACCAGTT CTCACAGTGA TTACTGTAGA 180
CTTCTCTGCG AATATATTCT TGGGAATGAT TTTACAGACC TTTTTGACAT TGTGATTACA 240
AATGCATTGA AGCCTGGTTT CTTCTCCCAC TTACCAAGTC AGAGACCTTT CCGGACACTC 300
GAGAATGATG AGGAGCAGGA GGCACTGCCA TCTCTGGATA AACCTGGCTG GTACTCCCAA 360
GGGAACGCTG TCCACCTCTA TGAACTTCTG AAGAAAATGA CTGGCAAACC TGAACCCAAG 420
GTTSTTTATT NWTGGTGWCA GCATGCAWTC AGATATTTTC CCAGCTCGTC ACTATAGTAA 480
TTGGGGAGAC AGTCCTCATC CGKGGAAGGA ACTCAGAGGG GGATGAARGG GCACGAGGGA 540
GTTCAGAGGC CTTGAGGGAG TTCAGAGCCT CTTAGAAGAA GGAAAGGGAA ATTTTGAGGG 600
GACCAAAAGN CAAAACCTTT AATTATTTCA TTTTAAANAT GGGGGTTTTT TTTTN 655 (2) INFORMATION FOR SEQ ID NO 27 (l) SEQUENCE CHARACTERISTICS
(A) LENGTH. 199 ammo acids
(B) TYPE: amino acid
(C) STRANDEDNESS:
(D) TOPOLOGY: linear
I n ) MOLECULE TYPE : protein
(xi f SEQUENCE DESCRI PTION . SEQ ID NO : 27 :
Lys Glu He Gin Ala Asp He Tyr He Val Val Pro Glu Ser Val Lys
1 5 10 15
Lys Trp Leu Arg Gin Leu Lys Asn Ala Gly Lys He Leu Leu Leu Xaa 20 25 30
Thr Ser Ser His Ser Asp Tyr Cys Arg Leu Leu Cys Glu Tyr He Leu 35 40 45
Gly Asn Asp Phe Thr Asp Leu Phe Asp He Val He Thr Asn Ala Leu 50 55 60
Lys Pro Gly Phe Phe Ser His Leu Pro Ser Gin Arg Pro Phe Arg Thr 65 70 75 80
Leu Glu Asn Asp Glu Glu Gin Glu Ala Leu Pro Ser Leu Asp Lys Pro 85 90 95
Gly Trp Tyr Ser Gin Gly Asn Ala Val His Leu Tyr Glu Leu Leu Lys 100 105 110
Lys Met Thr Gly Lys Pro Glu Pro Lys Val Xaa Tyr Xaa Trp Xaa Gin 115 120 125
His Ala Xaa Arg Tyr Phe Pro Ser Ser Ser Leu Xaa Xaa Leu Gly Arg 130 135 140
Gin Ser Ser Ser Xaa Glu Gly Thr Gin Arg Gly Met Lys Gly His Glu 145 150 155 160
Gly Val Gin Arg Pro Xaa Gly Ser Ser Glu Pro Leu Arg Arg Arg Lys 165 170 175
Gly Lys Phe Xaa Gly Asp Gin Lys Xaa Lys Pro Leu He He Ser Phe 180 185 190
Xaa Xaa Trp Gly Phe Phe Phe 195
(2) INFORMATION FOR SEQ ID NO:28:
(l) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 279 base pairs IB) TYPE: nucleic acid (C) STRANDEDNESS: double ID) TOPOLOGY: linear
111) MOLECULE TYPE: cDNA (xi) SEQUENCE DESCRIPTION: SEQ ID NO:28:
TCCTCCACTG NTCTTATCAA GTGATGAGAC ACTGATATCC AAATAANTNG TATTTACTGA 60
AAAATGAAGT GAAGACCCAT ATATGCAGTT AAAAAAAAGT TAATTTTCAA AAAATACTGT 120
AAAAGACTTT AAGGAACAAG TTTTATTGAC CAATAAGTTG ATATTTGTCC ATAGGTCTCC 180
TTTCTATAAA TCATCTTGAT GTTTAACAAC TCTTATTATA TTAAAATCTC AGTATCCTAA 240
AACTTAAAAA AAAAAAAAAA AAAAACATGT TTAATTAAK 279 (2) INFORMATION FOR SEQ ID NO:29:
(l) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 391 base pairs
(B) TYPE: nucleic acid
(C) STRANDEDNESS: double
(D) TOPOLOGY: linear
In) MOLECULE TYPE: cDNA
(xi) SEQUENCE DESCRIPTION: SEQ ID NO:29:
GAACNTGGGC CGCATGTATN TCTTCTATGG CAACAAGACC TCGGTGCAGT TCCAGAATTT 60
CTCACCCACT GTGGTTCACC CGGGAGACCT CCAGACTCAG CTGGCTGTGC AGACCAAGCG 120
CGTGGCGGCG CAGGTGGACG GCGGCGCGCA GGTGCAGCAG GTGCTCAATA TCGAGTGCCT 180
GCGGGACTTC CTGACGCCCC CGCTGCTGTC CGTGCGCTTC CGGTACGGTG GCGCCCCCCA 240
GGCCCTCACC CTGAAGCTCC CAGTGACCAT CAACAAGTTC TTCCAGCCCA CCGAGATGGC 300
GGCCCAGGAT TTCTTCCAGC GCTGGAAGCA GCTGANCCTC CCTCAACAGG AGGCGCAGAA 360
AATCTTCAAA GCCAACCACC CCATGGACGC A 391 (2) INFORMATION FOR SEQ ID NO:30:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 126 amino acids
(B) TYPE: ammo acid
(C) STRANDEDNESS:
(D) TOPOLOGY: linear
(ii) MOLECULE TYPE: protein
(xi) SEQUENCE DESCRIPTION: SEQ ID NO:30:
Met Tyr Xaa Phe Tyr Gly Asn Lys Thr Ser Val Gin Phe Gin Asn Phe 1 5 10 15
Ser Pro Thr Val Val His Pro Gly Asp Leu Gin Thr Gin Leu Ala Val 20 25 30
Gin Thr Lys Arg Val Ala Ala Gin Val Asp Gly Gly Ala Gin Val Gin 35 40 45
Gin Val Leu Asn He Glu Cys Leu Arg Asp Phe Leu Thr Pro Pro Leu 50 55 60
Leu Ser Val Arg Phe Arg Tyr Gly Gly Ala Pro Gin Ala Leu Thr Leu 65 70 75 80
Lyl Leu Pro Val Thr He Asn Lys Phe Phe Gin Pro Thr Glu Met Ala 85 90 95
Ala Gin Asp Phe Phe Gin Arg Trp Lys Gin Leu Xaa Leu Pro Gin Gin 100 105 110
Glu Ala Gin Lys He Phe Lys Ala Asn His Pro Met Asp Ala 115 120 125
(2) INFORMATION FOR SEQ ID NO.31.
(l) SEQUENCE CHARACTERISTICS
(A) LENGTH: 197 base pairs
(B) TYPE: nucleic acid
(C) STRANDEDNESS: double ID) TOPOLOGY- linear
In) MOLECULE TYPE: cDNA
(xi) SEQUENCE DESCRIPTION SEQ ID NO:31
CCCCTCNTCC NTTTCCCCCC CAAGCACAGA GGGGAGAGGG GCCAGGGAAG TGGATGTTTC 60
TTCCCNTCCC ACCCCACCCT GTTGTAGCCC CTCCTACCCC CTCCCCATCC AGGGGCTGTG 120
TATTATTGTG AGCGNATAAA CAGAGAGACG CTAAAAAAAA AAAAAAAAAA AAAAAAATCC 180
NNTAATTAAG CGGCCGC 197
(2) INFORMATION FOR SEQ ID NO.32. (l) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 514 base pairs
(B) TYPE: nucleic acid
(C) STRANDEDNESS: double ID) TOPOLOGY: linear
(ii) MOLECULE TYPE: cDNA
(xi) SEQUENCE DESCRIPTION: SEQ ID NO:32:
AAGCTTGGCA CGTGGCTGAT TGGAGCTGTA AAACATCATC AGGTGTTGCT ATTWTTTTAT 60
ATGATTATTC TGTTACTTGT ATTTATTGTT CAGTTTTCTG TATCTTGCGC TTGTTTAGCC 120
CTGAACCAGG AGCAACAGGG TCAGCTTCTG GAGGTTGGTT GGAACAATAC GGCAAGTGCT 180 CGAAATGACA TCCAGAGAAA TCTAAACTGC TGTGGGTTCC GAAGTGTTAA CCCAAATGAC 240
ACCTGTCTGG CTAGCTGTGT TAAAAGTGAC CACTCGTGCT CGCCATGTGC TCCAATCATA 300
GGAGAATATG CTGGAGAGGT TTTGAGATTT GTTGGTGGCA TTGGCCTGTT CTTCAGTTTT 360
ACAGAGATCC TGGGGTGTTT GGCTGACCTA CAGATACAGG AACCAGAAAG ACCCCCGCGC 420
GAATCCTAGT GCATTCCTTT GGATGAGGAA AACAAGGGAA GNTTCCNTTT CGTATTATGG 480 NCTTGTTTCA CTTTCTGTAA TTTTTCTGTT AAGG 514
(2) INFORMATION FOR SEQ ID NO: 33:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 151 amino acids
(B) TYPE: amino acid
(C) STRANDEDNESS:
(D) TOPOLOGY: linear
(ii) MOLECULE TYPE: protein
(xi) SEQUENCE DESCRIPTION: SEQ ID NO: 33:
Met He He Leu Leu Leu Val Phe He Val Gin Phe Ser Val Ser Cys
1 5 10 15
Ala Cys Leu Ala Leu Asn Gin Glu Gin Gin Gly Gin Leu Leu Glu Val 20 25 30
Gly Trp Asn Asn Thr Ala Ser Ala Arg Asn Asp He Gin Arg Asn Leu 35 40 45
Asn Cys Cys Gly Phe Arg Ser Val Asn Pro Asn Asp Thr Cys Leu Ala 50 55 60
Ser Cys Val Lys Ser Asp His Ser Cys Ser Pro Cys Ala Pro He He
65 70 75 80
Gly Glu Tyr Ala Gly Glu Val Leu Arg Phe Val Gly Gly He Gly Leu 85 90 95
Phe Phe Ser Phe Thr Glu He Leu Gly Cys Leu Ala Asp Leu Gin He 100 105 110
Gin Glu Pro Glu Arg Pro Pro Arg Glu Ser Xaa Cys He Pro Leu Asp 115 120 125
Glu Glu Asn Lys Gly Xaa Phe Xaa Phe Val Leu Trp Xaa Cys Phe Thr 130 135 140
Phe Cys Asn Phe Ser Val Lys 145 150
(2) INFORMATION FOR SEQ ID NO: 34:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 218 base pairs
(B) TYPE: nucleic acid (C) STRANDEDNESS: double
(D) TOPOLOGY, linear
(11) MOLECULE TYPE. cDNA
(xi) SEQUENCE DESCRIPTION. SEQ ID NO:34:
ACGTAGCAAA AAGATATTTG ATTATCTTAA AAATTGTTAA ATACCGTTTT CANGAAAGTT 60
CTCAGTATTG TAACAGCAAC TTGTCAAACC TAAGCATATT TGAATNTGAT NTCCCATAAT 120
TTGAAATNGA AATCGTATGG TGTGGCTCTG TATATTCTGT TAAAAAATTA AGGGACCAGA 180
AACCTTAAAA AAAAAAAAAA AAAATTCCCT GCGGCCGC 218
(2) INFORMATION FOR SEQ ID NO:35. (i) SEQUENCE CHARACTERISTICS:
(A) LENGTH. 525 base pairs
(B) TYPE: nucleic acid
(C) STRANDEDNESS. double
(D) TOPOLOGY, linear
In) MOLECULE TYPE: cDNA
(xi) SEQUENCE DESCRIPTION. SEQ ID NO: 35:
CAAGATTGGC AAGATGCTTA TTTNTGNNNC CATATTTGGC TGCCTTGACC CAGTGGCAAC 60
ACTAGCTGCA GTTATGACAG AGAAGTCTCC TTTTACCACA CCAATTGGTC GAAAAGATGA 120
AGCAGATCTT GCAAAATCAG CTTTGGCCAT GGCGGATTCA GACCACCTGA CGATCTACAA 180
TGCATATCTA GGATGGAAAG AAAGCACGAC AAGAAGGAGG TTATCGTTCT GAAATCACAT 240
ACTGCCGGAG GNAACTTTCT TAATANAACA TCACTGTTAA CCCTAGAGGA TGTAAAGCAG 300
GAGTTAATAA AGTTGGTTAA GGCAGCAGGA TTTTCATCTT CCACAACTTC TACCAGCTGG 360
GAAGGAAACA GANCCTCACA GACCCTCTCA TTCCAAGAAA TTGCCCTTCT TAAANCTGTA 420
CTGGTGGCTG GACTGTATGA CAATGTNGGG AAAATAATCT ATACAAATCN NTGGATGTTA 480
CANAAAAATT GGCTTGCATT GTGGANACGG CCCAGGCNAA ACACA 525 (2) INFORMATION FOR SEQ ID NO:36:
(l) SEQUENCE CHARACTERISTICS:
(A) LENGTH. Ill amino acids
(B) TYPE: ammo acid
(C) STRANDEDNESS:
ID) TOPOLOGY: linear
In) MOLECULE TYPE: protem (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 36.
Met Glu Arg Lys His Asp Lys Lys Glu Val He Val Leu Lys Ser His 1 5 10 15
Thr Ala Gly Gly Asn Phe Leu Asn Xaa Thr Ser Leu Leu Thr Leu Glu 20 25 30
Asp Val Lys Gin Glu Leu He Lys Leu Val Lys Ala Ala Gly Phe Ser 35 40 45
Ser Ser Thr Thr Ser Thr Ser Trp Glu Gly Asn Arg Xaa Ser Gin Thr 50 55 60
Leu Ser Phe Gin Glu He Ala Leu Leu Lys Xaa Val Leu Val Ala Gly 65 70 75 80
Leu Tyr Asp Asn Val Gly Lys He He Tyr Thr Asn Xaa Trp Met Leu 85 90 95
Xaa Lys Asn Trp Leu Ala Leu Trp Xaa Arg Pro Arg Xaa Asn Thr 100 105 110
(2) INFORMATION FOR SEQ ID NO: 37
(l) SEQUENCE CHARACTERISTICS-
(A) LENGTH: 109 base pairs IB) TYPE- nucleic acid
(C) STRANDEDNESS: double
(D) TOPOLOGY: linear
111) MOLECULE TYPE: cDNA
(xi ) SEQUENCE DESCRIPTION: SEQ ID NO: 37: ACATGTATAA TTTTNTAGTT TCCTTTTTAA TGATGATTAT TCTGAATGTA TTTGCCANTA 60 CANNTACAAT AAATTTNTTT GGTATTATGC AAAAAAAAAA AAAAAAANA 109
(2) INFORMATION FOR SEQ ID NO:3B:
(l) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 825 base pairs
(B) TYPE: nucleic acid
(C) STRANDEDNESS: double
(D) TOPOLOGY: linear
111) MOLECULE TYPE: cDNA
(xi) SEQUENCE DESCRIPTION: SEQ ID NO: 38:
GAATTCGGCA CGAGTTTTTT TTTTCTGCAG TTGTGTGTAT GTGTGTTTGT GTGAAGAAAA 60
ACAGACTCTG TCCAGGTAGA AATGGTGAGG AGGGGGAAGA GAATTACATT TCCAGGGTCA 120
GAAACTTGGC AACAGTTTTC CTAKAGTGAC TCAGACACAC CACAGTAACA ACTCTCGCTG 180 CAATTTTATT TTAATTTGAG AAATAAAGAT TTCCTCCAAG CCACATGAGG ACTCTGGCAC 240
CCACCCACAA AGCAAGACCT GTATTTATAA GCCGAGGGTG CAGGGAGCTN AACTGCGGGA 300
CCCGTCAGGG CCCCGTGGAC CCATCCCCGT CCCCACCCCC CCCTCCACCG YTGGGGCCCA 360
TCAGTGTGTG TTGGGGGGGA TGCTTGGGCA GCTGGGGGGT GAGGGAGACA ACAAACCTYG 420
GGGAAYTGGG AGCCAGAGCT GCGGCCTGAC TGACGCCTTT TGATGCTCAC GGGAAATTTN 480
TGCCCAGGAT NTCAGCCCCA GGCTGGTTGT TTCTACAAAT CTCTCTCAAA TGTATTATTT 540
TGGTGACAAA AATGAAGGAG CTTTGTAAAT TTTTTTAAAA TTATGAATNC ATATCAAGTA 600
GTTGTTTACA TTTCTTGAAA AAATAGGAAC TCGGGCAGCA GAATCAGATT GGCAGAATCT 660
TTAGACTACA CAGGCAATAA TCAAGTCTGC TGTTTTGNCC TTTCGTAGTA GAAGTGGTTG 720
TAGTGTTTAG ATATCTGTTT GGTCTTGCTT CTTGTATTGC ATTTTTTTCA ATAAACAACA 780
ACAAAAAGAA AAAAAAAAAA AAAAAAAAAA AAGATCTTTA ATTAA 825 (2) INFORMATION FOR SEQ ID NO : 39 :
(l) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 200 amino acids
Figure imgf000097_0001
(C) STRANDEDNESS:
(D) TOPOLOGY, linear
(ii) MOLECULE TYPE: protein
(xi ) SEQUENCE DESCRIPTION: SEQ ID NO: 39:
Met Arg Thr Leu Ala Pro Thr His Lys Ala Arg Pro Val Phe He Ser
1 5 10 15
Arg Gly Cys Arg Glu Leu Asn Cys Gly Thr Arg Gin Gly Pro Val Asp 20 25 30
Pro Ser Pro Ser Pro Pro Pro Pro Pro Pro Leu Gly Pro He Ser Val 35 40 45
Cys Trp Gly Gly Cys Leu Gly Ser Trp Gly Val Arg Glu Thr Thr Asn 50 55 60
Leu Gly Glu Leu Gly Ala Arg Ala Ala Ala Xaa Leu Thr Pro Phe Asp 65 70 75 80
Ala His Gly Lys Phe Xaa Pro Arg Xaa Ser Ala Pro Gly Trp Leu Phe 85 90 95
Leu Gin He Ser Leu Lys Cys He He Leu Val Thr Lys Met Lys Glu 100 105 HO
Leu Cys Lys Phe Phe Xaa Asn Tyr Glu Xaa He Ser Ser Ser Cys Leu 115 120 125
His Phe Leu Lys Lys Xaa Glu Leu Gly Gin Gin Asn Gin He Gly Arg 130 135 140
He Phe Arg Leu His Arg Gin Xaa Ser Ser Leu Leu Phe Xaa Pro Phe 145 150 155 160
Val Val Glu Val Val Val Val Phe Arg Tyr Leu Phe Gly Leu Ala Ser 165 170 175
Cys He Ala Phe Phe Ser He Asn Asn Asn Lys Lys Lys Lys Lys Lys 180 185 190
Lys Lys Lys Lys Asp Leu Xaa Leu 195 200
(2) INFORMATION FOR SEQ ID NO: 40:
(l) SEQUENCE CHARACTERISTICS:
(A) LENGTH- 508 base pairs
(B) TYPE: nucleic acid
(C) STRANDEDNESS: double
(D) TOPOLOGY, linear
(11) MOLECULE TYPE: cDNA
(xi) SEQUENCE DESCRIPTION: SEQ ID NO: 40 •
AAGCTTGGCA CGNGGTCTGT CGCTCCCGGA AACTTGTTGG CAATGCCTAT TTTTTGGCTT 60
TCCCCCGCGT TCTCTAAACT AACTATTTAA AGGTCTGCGG TCCCSAAATG GTTTGACTAA 120
ACGTAGGATG GGACTTAAGT TGAACGGCAG ATATATTTCA CTGATCCTCG CGGTGCAAAT 180
AGCGTATCTG GTGCAGGCCG TGAGAGCAGC GGGCAAGTGC GATGCGGTCT TCAAGGGCTT 240
TTCGGACTGT TTGCTCAAGC TGGGCGAMMR CΛTGGGCCAA CTACCCGCAG GSCTKGGACG 300
ACAAGACGAA CATCAAGACC GTGTGCACAT ACTGGGAGGA TTTCCACAGC TGCACGGTCA 360
CAGCCCTTAC GGATTGCCAG GGAAGGGGCG AAAGATATGT GGGGATAAAC TGAGAAAAGA 420
ATCCAAAAAC CTCAACATCC AAGGGCAGCT TATTTCGAAY TYTGCGGCAN GTCAACGGNG 480 GCGGCCGGGT CCTTGTTCCC GGCTTTTT 508
(2) INFORMATION FOR SEQ ID NO: 41:
(l) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 127 ammo acids
(B) TYPE: amino acid
(C) STRANDEDNESS:
(D) TOPOLOGY- linear
(ii) MOLECULE TYPE: protem
(xi) SEQUENCE DESCRIPTION: SEQ ID NO : 41 : Met Gly Leu Lys Leu Asn Gly Arg Tyr He Ser Leu He Leu Ala Val 1 5 10 15
Gin He Ala Tyr Leu Val Gin Ala Val Arg Ala Ala Gly Lys Cys Asp 20 25 30
Ala Val Phe Lys Gly Phe Ser Asp Cys Leu Leu Lys Leu Gly Xaa Xaa 35 40 45
Met Gly Gin Leu Pro Ala Gly Leu Gly Arg Gin Asp Glu His Gin Asp 50 55 60
Arg Val His He Leu Gly Gly Phe Pro Gin Leu His Gly His Ser Pro 65 70 75 80
Tyr Gly Leu Pro Gly Lys Gly Arg Lys He Cys Gly Asp Lys Leu Arg 85 90 95
Lys Glu Ser Lys Asn Leu Asn He Gin Gly Gin Leu He Ser Asn Xaa 100 105 110
Ala Ala Xaa Gin Arg Xaa Arg Pro Gly Pro Cys Ser Arg Leu Phe 115 120 125
(2) INFORMATION FOR SEQ ID NO 42
(l) SEQUENCE CHARACTERISTICS.
(A) LENGTH. 269 base pairs
(B) TYPE nucleic acid
(C) STRANDEDNESS double
(D) TOPOLOGY: linear
In) MOLECULE TYPE cDNA
(Xl) SEQUENCE DESCRIPTION SEQ ID NO -42:
TGGTTTTAGC TGTTACACAC ACAGTAATAC CTGAATATCC CACGGTATAG ATCACANGGG 60
GGGGATGTTA AATGTTAATC TAAAATATAG CTAAAAAAAG ATTTTGACAT AAAAGAGCCT 120
TGATTTTAAA AAAAAAAGAG AGAGAGATGT AATTTAAAAA GTTTATTATA AATTAAATTC 180
AGCNAAAAAA GATTTGCTAC AAAGTATAGA GAAGTATAAA ATAAAAGTTA TTGTTTGNAA 240
AAAAAAAAAA AAAAATTNCC TGCGGCCGC 269 (2) INFORMATION FOR SEQ ID NO:43:
(l) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 676 base pairs
(B) TYPE: nucleic acid
(C) STRANDEDNESS: double
(D) TOPOLOGY: linear
(n) MOLECULE TYPE: cDNA (xi) SEQUENCE DESCRIPTION: SEQ ID NO:43.
GAGATTTTCA GCACCCTGCG ATATGCAAGC CGAGNTCAGC GGGTCACCAC CCGACCACAG 60
GCCCCCAACT TTCCTGTGGC AAAGCAGCCC CAGCGTTTGG AGACAGAGAT GCTGCAGCTC 120
CAGGAGGAGA ACCGTCGCCT GCAGTTCCAG NTGGACCAAA TGGANTGCAA GGCCTCAGGG 180
TTCAGTGGAG CCCGGGTGGC CTGGGCCCAG CGGAACCTGT ACGGGATGNT ACAGGAGTTT 240
CATGNTAGAG AATGAGAGGC TCAGGAAAGA AAAGAGCCAG CTGCAGAATA GCCGAGAGCT 300
AGCCCAGAAT GAGCAGCGCA TCCTGGCCCA GCAGGTCCAT GCACTAGAGA RGCGTCTCCT 360
CTCTGCCTGC TACCATCACC AGCAGGGTCC TGGCCTGACC CCACCGTGTC CCTGCTTGAT 420
GGCCCCAGCT CCCCCTTGCC ATGCACTGCC ACCCCTCTAC TCCTGCCCCT GCTGCCACAT 480
CTGCCCACTG TGTCKAGTGC CCCTGGCCCA CTGGGYYKGC CTGSCMAGGG GAGCACCACC 540
TTGCCCCAGC CTCTCTTCTG GGGCTCTGAR GAGTCAGAAA TAGACCAGAC GTGGTTTCCT 600
GGTTCTCAGG ANGGTTTTTA GTTTNΛGGAG AGGGACGGTA GAAGAACCAT TTTGTTGCAA 660
AAAGAAGGGG ACCAAG 676 (2) INFORMATION FOR SEQ ID NO:44:
(l) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 218 amino acids
(B) TYPE: amino acid
(C) STRANDEDNESS:
(D) TOPOLOGY, linear
(li) MOLECULE TYPE: protein
(xi) SEQUENCE DESCRIPTION: SEQ ID NO: 44 :
Met Gin Ala Glu Xaa Ser Gly Ser Pro Pro Asp His Arg Pro Pro Ser 1 5 10 15
Phe Leu Trp Gin Ser Ser Pro Ser Val Trp Arg Gin Arg Cys Cys Ser 20 25 30
Ser Arg Arg Arg Thr Val Ala Cys Ser Ser Xaa Trp Thr Lys Trp Xaa 35 40 45
Ala Arg Pro Gin Gly Ser Val Glu Pro Gly Trp Pro Gly Pro Ser Gly 50 55 60
Thr Cys Thr Gly Xaa Tyr Arg Ser Phe Met Xaa Glu Asn Glu Arg Leu 65 70 75 80
Arg Lys Glu Lys Ser Gin Leu Gin Asn Ser Arg Glu Leu Ala Gin Asn 85 90 95
Glu Gin Arg He Leu Ala Gin Gin Val His Ala Leu Glu Xaa Arg Leu 100 105 110 leu Ser Ala Cys lyr His His Gin Gin Gly Pro Gly Leu Thr Pro Pro 115 120 125
Cys Pro Cys Leu Met Ala Pro Ala Pro Pro Cys His Ala Leu Pro Pro 130 135 140
Leu Tyr Ser Cys Pro Cys Cys His He Cys Pro Leu Cys Xaa Val Pro 145 150 155 160
Leu Ala His Trp Xaa Xaa Leu Xaa Arg Gly Ala Pro Pro Cys Pro Ser 165 170 175
Leu Ser Ser Gly Ala Leu Xaa Ser Gin Lys Xaa Thr Arg Arg Giv Phe 180 185 190
Leu Val Leu Arg Xaa Val Phe Ser Xaa Arg Arg Gly Thr Val Glu Glu 195 200 205
Pro Phe Cys Cys Lys Lys Lys Gly Thr Lys 210 215
(2) INFORMATION FOR SEQ ID NO 45
(l) SEQUENCE CHARACTERISTICS
(A) LENGTH 394 base pairs
(B) TYPE nucleic acid
(C) STRANDEDNESS double
(D) TOPOLOGY linear
In) MOLECULE TYPE cDNA
(XI) SEQUENCE DESCRIPTION SEQ ID NO 45
TTCCAAACTT GGCCCAGAGA CTGGAGGCCT TCAGAGACCA GATTGGCAGN TCCNTGCGAN 60
GTGGCCGCAG CCAGCCACCC TGCAGTGAGG GCGCACGGAG CNCAGGCCAA GTCNTCCNTC 120
CCCATTGAAG GCCAAGTGGG AACNNANNAG AATGCTGTGT GACCTCAGAC TGGGCTCCAC 180
ACTCTTGGGC TTCAGTCTGC CCATCTGCTG AATGGAGACA GCAGCTGNTA CTCCACCTGC 240
AGCTGGGCTA GGGGCGGGGA CTGGGGGTGC TATTTAGGGG AACAAGGGGA TTTCAGGAG^ 300
AACCCAGGCA GCAGGGGATG AAATACATGA ATAAAGAGAG GCATCAGCTC CAAAAAAAAA 360
AAAAAAAAAA AAAGAACTTT AATTAAGCGG CCGC 394 (2) INFORMATION FOR SEQ ID NO 46
(l) SEQUENCE CHARACTERISTICS
(A) LENGTH 479 base pairs
(B) TYPE: nucleic acid
(C) STRANDEDNESS double ID) TOPOLOGY linear
(u) MOLECULE TYPE- cDNA (xi) SEQUENCE DESCRIPTION. SEQ ID NO: 46-
AAGCTTGGCA CNAGGGCCAA ACCTCTATGG ATATATAAAG GGAAGCTTGA GGAGGAATTT 60
CACAGTTACA GTGCAGAAGC AGAAGCAAAA GAATTAACCA GCTCTTCAGT CAAGCAAATC 120
CTCTACTCAC CATGCTTCCT CCTGCCATTC ATTTCTATCT CCTTCCCCTT GCATGCATCC 180
TAATGAAAAG CTGTTTGGCT TTTAAAAATG ATGCCACAGA AATCCTTTAT TCACATGTGG 240
TTAAACCTGT TCCAGCACAC CCCAGCAGCA ACAGCACGTT GAATCAAGCC AGAAATGGAG 300
GCAGGCATTT CAGTAACACT GGACTGGATC GGAACACTCG GGTTCAAGTG GGTTGCCGGG 360
AACTGCGTTC CACCAAATAC ATCTCTGGAT GGGCCAGTTG CACCAGCATT CAGCCCTCTG 420
GAAGGGAGCT GGGTGTGTGG TGGGCGAGTG CTTTGCCCNT GCCAGTGGTT CCCTAACTG 479 (2) INFORMATION FOR SEQ ID NO:47:
(l) SEQUENCE CHARACTERISTICS.
(A) LENGTH: 116 amino acids
(B) TYPE: amino acid
(C) STRANDEDNESS.
(D) TOPOLOGY: linear
(ii) MOLECULE TYPE: protein
(xi) SEQUENCE DESCRIPTION: SEQ ID NO: 47-
Met Leu Pro Pro Ala He His Phe Tyr Leu Leu Pro Leu Ala Cys He 1 5 10 15
Leu Met Lys Ser Cys Leu Ala Phe Lys Asn Asp Ala Thr Glu He Leu 20 25 . 30
Tyr Ser His Val Val Lys Pro Val Pro Ala His Pro Ser Ser Asn Ser 35 40 45
Thr Leu Asn Gin Ala Arg Asn Gly Gly Arg His Phe Ser Asn Thr Gly 50 55 60
Leu Asp Arg Asn Thr Arg Val Gin Val Gly Cys Arg Glu Leu Arg Ser 65 70 75 80
Thr Lys Tyr He Ser Gly Trp Ala Ser Cys Thr Ser He Gin Pro Ser 85 90 95
Gly Arg Glu Leu Gly Val Trp Trp Ala Ser Ala Leu Pro Xaa Pro Val 100 105 110
Val Pro Xaa Leu 115
(2) INFORMATION FOR SEQ ID NO:48:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 35 base pairs
(B) TYPE: nucleic acid (C) STRANDEDNESS: double
(D) TOPOLOGY: linear
(11) MOLECULE TYPE: cDNA
(xi ) SEQUENCE DESCRIPTION: SEQ ID NO: 48: AAGTTTAAAA AAAAAAAAAA AAATCNCGCG GCCGC 35 (2) INFORMATION FOR SEQ ID NO:49:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 296 base pairs
(B) TYPE: nucleic acid
(C) STRANDEDNESS: double
(D) TOPOLOGY: linear
111) MOLECULE TYPE: cDNA
(xi) SEQUENCE DESCRIPTION: SEQ ID NO : 49 : ACATTTACTT AAAGGAGAAA AGTAAGGGGG TCNCAGAAAT GTCTGGGGCN ATTATAGAAA 60 ACATGAGTAC CAAGAAGCTC TGCATTGTTG GAGGGATTCT TCTGGTTTTC CCAATCGTTG 120 CCTNTCTGGT GGGAGGCTTG ATCGCTCCAG CACCCACAAC ANCAGTACCC TACACGTCAA 180 TAAAATGTGT GGATGTCCGT AAGAACCACC ATAAAACAAG ATGACTGGCT CCTTGGGGAC 240 CTAACAAGTG TTTNCAGACC CATCNNTNAG CCGAACAAAC ANCCAGCGCC AATGTA 296 (2) INFORMATION FOR SEQ ID NO: 50:
(l) SEQUENCE CHARACTERISTICS:
(A) LENGTH. 332 base pairs
(B) TYPE: nucleic acid
(C) STRANDEDNESS: double
(D) TOPOLOGY: linear
111) MOLECULE TYPE: cDNA
(xi ) SEQUENCE DESCRIPTION: SEQ ID NO: 50:
GAATTCGGCA CGAGCTTGAT TGCTCCAGGG CCCACAACGG CAGTGTCCTA CATGTCGGTG 60
AAATGTGTGG ATGCCCGTAA GAACCATCAC AAGACAAAAT GGTTCGTGCC TTGGGGACCC 120
AATCATTGTG ACAAGATCCG AGACATTGAA GAGGCAATTC CAAGGGAAAT TGAAGCCAAT 180
GACATCGTGT TTTCTGTTCA CATTCCCCTC CCCCACATGG GAGATGAGTC CTTGGTTCCA 240
ATTCATGMTG TTTATCCTGG CAGCTGGGAC ATTGCCTTTC AAGCTAAACA ACCAAATCAG 300 GGGAAAATGC AGGAAGTCTC CATGGGACGT TT 332
(2) INFORMATION FOR SEQ ID NO:51:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 110 ammo acids
(B) TYPE: ammo acid
(C) STRANDEDNESS:
(D) TOPOLOGY: linear
In) MOLECULE TYPE: protein
(Xl ) SEQUENCE DESCRIPTION: SEQ ID NO: 51:
Glu Phe Gly Thr Ser Leu He Ala Pro Gly Pro Thr Thr Ala Val Ser
1 5 10 15
Tyr Met Ser Val Lys Cys Val Asp Ala Arg Lys Asn His His Lys Thr 20 25 30
Lys Trp Phe Val Pro Trp Gly Pro Asn His Cys Asp Lys He Arg Asp 35 40 45
He Glu Glu Ala He Pro Arg Glu He Glu Ala Asn Asp He Val Phe 50 55 60
Ser Val His He Pro Leu Pro His Met Gly Asp Glu Ser Leu Val Pro 65 70 75 80
He His Xaa Val Tyr Pro Gly Ser Trp Asp He Ala Phe Gin Ala Lys 85 90 95
Gin Pro Asn Gin Gly Lys Met Gin Glu Val Ser Met Gly Arg 100 105 110
(2) INFORMATION FOR SEQ ID NO: 52.
(l) SEQUENCE CHARACTERISTICS.
(A) LENGTH: 327 base pairs
(B) TYPE: nucleic acid
(C) STRANDEDNESS: double
(D) TOPOLOGY: linear
(il) MOLECULE TYPE: cDNA
(xi) SEQUENCE DESCRIPTION: SEQ ID NO:52:
TCACTCCTAA TCCATGACCA CTGTTTTTTT CCTATTTATA TCACCAGGTA GCCTACTGAG 60
TTAATATTTA AGTTGTCNNT GGGTNNGTGT CCCTGTTTTG TGGCATAATA TAACTGAATT 120
TCATGNGAAG ATTTATTCCA CCAGGGGTAT TTCAGCTTTG AAACCAAATC TGTGTATCTA 180
ATACTAACCA ATCTGTTGGA TGTGGATTTT AAAAAATGTT TGCTAAACTA CCCAAGTAAG 240
ATTTACTGTA TTAAATGGCC TTCGGGTCTG AAAAGCTTTT TTAAAAAAAA AAAAAAAAAA 300 AAAAAAAAAA AAAAGATCTT TAATTAA 327
( 2 ) INFORMATION FOR SEQ ID NO 53
( l ) SEQUENCE CHARACTERISTICS
(A) LENGTH 242 base pairs
( B ) TYPE nucleic acid
(C) STRANDEDNESS double
(D) TOPOLOGY l inear
(n) MOLECULE TYPE cDNA
(xi) SEQUENCE DESCRIPTION SEQ ID NO 53
GCGAAAGGAT TTTAAGGAAC AGATCATCCA CCATGTGGCC ACTATCATTC TCCTCTGCTT 60
CTCCTGGTTT GCCAATTACG TCCGGGCAGG GACCCTCATC ATGGCTCTGC ATGACGCTTC 120
TGACTACCTG CTGGAGTCTG CCAAGATGTT TAACTACGCG GGATGGAAGA ACACCTGCAA 180
CAACCTCTTC ATTGTGTTCG CCATCGTTTT CATCATCACT CGGCTGGTTA TCATGCCTTT 240
CT 242 (2) INFORMATION FOR SEQ ID NO 54
(l) SEQUENCE CHARACTERISTICS
(A) LENGTH 377 base pairs
(B) TYPE nucleic acid
(C) STRANDEDNESS double
(D) TOPOLOGY linear
111) MOLECULE TYPE cDNA
(Xl) SEQUENCE DESCRIPTION SEQ ID NO 54
GAATTCGGCA CGAGGATTCT CATCAGCTTT TCCTGGGTTT GCCAATTACA TCCGAGCTGG 60
GACTCTAATC ATGGCTCTGC ATGACTCTTC CGATTACCTG CTGGAKTCAG CCAAGATGTT 120
TAACTACGCG GGATGGAAGA ACACCTGCAA CAACATCTTC ATCGTCTTCG CCATTGTTTT 180
TATCATCACC CGACTGGTCA TCCTGCCCTT CTGGATCCTG CATTGCACCC TGGGTGTACC 240
CACTGGAGCT CTATCCTGCC TTCTTTGGGC TATTACTTCT TTCAATTCCA TGATGGGAGT 300
TCTACAGCTG CTGCATATCT TCTGGGSCTA CCTCATTTTG CGSATGGGCC CACAAGTTCA 360
TAACTGGGAA AGCTGGT 377
(2) INFORMATION FOR SEQ ID NO 55
(l) SEQUENCE CHARACTERISTICS
(A) LENGTH 102 amino acids IB) TYPE ammo acid (C) STRANDEDNESS I D ) TOPOLOGY , l inear ( i i ) MOLECULE TYPE : protem
( xi ) SEQUENCE DESCRI PTION : SEQ ID NO : 55 :
Met Ala Leu Hi s Asp Ser Ser Asp Tyr Leu Leu Xaa Ser Ala Lys Met
1 5 10 15
Phe Asn Tyr Ala Gly Trp Lys Asn Thr Cys Asn Asn He Phe He Val 20 25 30
Phe Ala He Val Phe He He Thr Arg Leu Val He Leu Pro Phe Trp 35 40 45
He Leu His Cys Thr Leu Gly Val Pro Thr Gly Ala Leu Ser Cys Leu 50 55 60
Leu Trp Ala He Thr Ser Phe Asn Ser Met Met Gly Val Leu Gin Leu 65 70 75 80
Leu His He Phe Trp Xaa Tyr Leu He Leu Arg Met Gly Pro Gin Val B5 90 95
Figure imgf000106_0001
100
(2) INFORMATION FOR SEQ ID NO:56: d) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 369 base pairs
(B) TYPE, nucleic acid
(C) STRANDEDNESS: double
(D) TOPOLOGY: linear
(ll) MOLECULE TYPE: cDNA
(xi) SEQUENCE DESCRIPTION: SEQ ID NO: 56:
AAAAAGTGGG GGCTGTACTG GGGACTGCTC GGATGATNTT TNTTAGTGNT ACTTTTTTCA 60
GCTGTCCCTG TAGCGACAGG TNTAAGATCT GACTGCCTCC TTTTTNTGGC NTCTTCCCCC 120
TTCCNTNTTC TCTTCAGNTA GGCTAGCTGG TTTGGAGTAG AATGGCAACT AATTNTAATT 180
TTTATTTATT AAATATTTGG GGTTTTGGTT TTAAAGCCAG AATTACGGNT AGCACCTAGC 240
ATTTCNGCAG AGGGACCATT TTNGACCNAA ATNTANTNTT NATGGGTTTT TTTTTAAAAT 300
TNAAAGATTA AATNNNAAAT ATTAAATAAA AAAAAAAAAA AAAAAAAAAA AAAAAAAAAG 360
CGCGGCCGC 369 (2) INFORMATION FOR SEQ ID NO: 57: (l) SEQUENCE CHARACTERISTICS. (A) LENGTH: 423 base pairs
(B) TYPE- nucleic acid
(C) STRANDEDNESS: double
(D) TOPOLOGY, linear
(n) MOLECULE TYPE: cDNA
(xi) SEQUENCE DESCRIPTION: SEQ ID NO: 57:
GAATTCGGCA CGNNGTGNAA TATAAAAATT TATTTTTAAG TCAAAGTATG CAACAAATAA 60
ACCTACAGAA AACATTTTCC CATCACAATC TGTTGCTTTA CCAAATAATA TTTTGAAAAC 120
ACATTCCTTC AGTCATTATA AAGTTCTTAA AATACAAAAG AAATTAAATC TGTAAGAAAG 180
TCTAGTAGAC CAGATGCTGT TGTCAAGACT TGTATGTTGG TGTTTTTGCT TTCAGTACAT 240
CCCACGCCAT CCACCTCCAC TYCATGCCGC CTTGCCCATA GTAACCTCCA CTGCCTCCAC 300
CACCACGGCC ATAACCACCC AAACCATCAG GAGTACCATA TCCTCCACTG TAATTGTTCC 360
CCATTCCCAT TCTTCCAACT GGATTCCATA GGCCYTCCCT GGATTATTTT TNAAAAGGAA 420
AAA 423 (2) INFORMATION FOR SEQ ID NO: 58:
(1) SEQUENCE CHARACTERISTICS .
(A) LENGTH. 76 amino acids
(B) TYPE: amino acid
(C) STRANDEDNESS:
(D) TOPOLOGY: linear
(n) MOLECULE TYPE: protein
(Xl ) SEQUENCE DESCRIPTION: SEQ ID NO: 58:
Met Leu Leu Ser Arg Leu Val Cys Trp Cys Phe Cys Phe Gin Tyr He 1 5 10 15
Pro Arg His Pro Pro Pro Leu His Ala Ala Leu Pro He Val Thr Ser 20 25 30
Thr Ala Ser Thr Thr Thr Ala He Thr Thr Gin Thr He Arg Ser Thr 35 40 45
He Ser Ser Thr Val He Val Pro His Ser His Ser Ser Asn Trp He 50 55 60
Pro Xaa Ala Xaa Pro Gly Leu Phe Xaa Lys Arg Lys 65 70 75
(2) INFORMATION FOR SEQ ID NO: 59:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 294 base pairs (B) TYPE: nucleic acid
(C) STRANDEDNESS: double
(D) TOPOLOGY: linear
111) MOLECULE TYPE. cDNA
(xi) SEQUENCE DESCRIPTION: SEQ ID NO:59:
TAAAAACtCC TTTTTCCTCN TANGGGTNTA TCATAGGGTC CCGGTNGCTG TCCCAGCAAT 60
TTTNTNGGNG GATCATAAAA TCCTTNGATT TNACTCGTGA NANTTGNGAA GATCTCAATA 120
TACCTATTTA AAAATGTTTT AAGGTACAGG TTTCAGCATA AATGTATTAG TGTAAATTAG 180
ATACNGGGCA AAATGCAGTA AGTTTTTNTA TATNTAGATA CATAACCCAA TTTAAATTGC 240
CTAAATACAC CGTAAGTTAA CAGTTTAAAC CTACAAACTT AATTAAGCGG CCGC 294

Claims

What is claimed is
1 A composition comprising an isolated protein encoded by a polynucleotide selected from the group consisting of
(a) a polynucleotide compπsing the nucleotide sequence of SEQ ID NO 1 ,
(b) a polynucleotide comprising the nucleotide sequence of SEQ ID NO 1 from nucleotide 70 to nucleotide 505,
(c) a polynucleotide comprising the nucleotide sequence of the full length protein coding sequence of clone API 62 deposited under accession number ATCC 98026 ,
(d) a polynucleotide encoding the full length protein encoded by the cDNA insert of clone API 62 deposited under accession number ATCC 98026 ,
(e) a polynucleotide comprising the nucleotide sequence of the mature protein coding sequence of clone API 62 deposited under accession number ATCC 98026 ,
(f) a polynucleotide encoding the mature protein encoded by the cDNA insert of clone AP162 deposited under accession number ATCC 98026 ,
(g) a polynucleotide encoding a protein comprising the amino acid sequence of SEQ ID NO 2,
(h) a polynucleotide encoding a protein comprising a fragment of the amino acid sequence of SEQ ID NO 2 having biological activity,
(i) a polynucleotide which is an allelic variant of a polynucleotide of (a)-(f) above, and
(j) a polynucleotide which encodes a species homologue of the protein of
(g) or (h) above
2 The composition of claim 1 , further compπsing a pharmaceutically acceptable carrier
3 A method for preventing, treating or ameliorating a medical condition which comprises administering to a mammalian subject a therapeutically effective amount of a composition of claim 2
4 A composition compπsing a protein, wherein said protein comprises an ammo acid sequence selected from the group consisting of
(a) the amino acid sequence of SEQ ID NO 2.
(b) the ammo acid sequence of SEQ ID NO 2 from amino acid 42 to amino ,
(c) fragments of the amino acid sequence of SEQ ID NO 2, and
(d) the amino acid sequence encoded by the cDNA insert of clone API62 deposited under accession number ATCC 98026, the protein being substantially free from other mammalian proteins
5 The composition of claim 4, wherein said protein comprises the amino acid sequence of SEQ ID NO 2
6 The composition of claim 4, wherem said protein comprises the ammo acid sequence of SEQ ID NO 2 from amino acid 42 to ammo acid 61
7 The composition of claim 2, further comprising a pharmaceutically acceptable carrier
8 A method for preventing, treating or ameliorating a medical condition which comprises administering to a mammalian subject a therapeuticallv effective amount of a composition of claim 7
9 A composition comprising an isolated protein encoded by a polynucleotide selected from the group consisting of
(a) a polynucleotide comprising the nucleotide sequence of SEQ ID NO 4,
(b) a polynucleotide comprising the nucleotide sequence of SEQ ID NO 4 from nucleotide 230 to nucleotide 791 ,
(c) a polynucleotide comprising the nucleotide sequence of SEQ ID NO 4 from nucleotide 311 to nucleotide 791 , (d) a polynucleotide comprising the nucleotide sequence of the full length protein coding sequence of clone AM931 deposited under accession number ATCC 98026 ,
(e) a polynucleotide encoding the full length protein encoded by the cDNA insert of clone AM931 deposited under accession number ATCC 98026 ,
(f) a polynucleotide comprising the nucleotide sequence of the mature pYotein coding sequence of clone AM931 deposited under accession number ATCC 98026 ,
(g) a polynucleotide encoding the mature protein encoded by the cDNA insert of clone AM931 deposited under accession number ATCC 98026 ,
(h) a polynucleotide encoding a protein comprising the amino acid sequence of SEQ ID NO 5,
(l) a polynucleotide encoding a protein comprising a fragment of the amino acid sequence of SEQ ID NO 5 having biological activity,
(j) a polynucleotide which is an allelic variant of a polynucleotide of (a)-(g) above, and
(k) a polynucleotide which encodes a species homologue of the protein of (h) or (l) above
10 A composition comprising a protein, wherein said protem comprises an ammo acid sequence selected from the group consisting of
(a) the amino acid sequence of SEQ ID NO 5,
(b) the amino acid sequence of SEQ ID NO 5 from ammo acid 32 to ammo acid 51 ,
(c) fragments of the amino acid sequence of SEQ ID NO 5, and
(d) the amino acid sequence encoded by the cDNA insert of clone AM931 deposited under accession number ATCC 98026, the protein being substantially free from other mammalian proteins
11 A composition comprising an isolated protein encoded by a polynucleotide selected from the group consisting of
(a) a polynucleotide comprising the nucleotide sequence of SEQ ID NO 6,
(b) a polynucleotide comprising the nucleotide sequence of SEQ ID NO 6 from nucleotide 14 to nucleotide 491 , (c) a polynucleotide comprising the nucleotide sequence of SEQ ID NO 6 from nucleotide 83 to nucleotide 491 ,
(d) a polynucleotide comprising the nucleotide sequence of the full length protein coding sequence of clone AM610 deposited under accession number ATCC 98026 ,
(e) a polynucleotide encoding the full length protein encoded by the cDNA Insert of clone AM610 deposited under accession number ATCC 98026 ,
(f) a polynucleotide comprising the nucleotide sequence of the mature protein coding sequence of clone AM610 deposited under accession number ATCC 98026 ,
(g) a polynucleotide encoding the mature protein encoded by the cDNA insert of clone AM610 deposited under accession number ATCC 98026 ,
(h) a polynucleotide encoding a protein comprising the amino acid sequence of SEQ ID NO 7,
(I) a polynucleotide encoding a protein comprising a fragment of the amino acid sequence of SEQ ID NO 7 having biological activity,
(j) a polynucleotide which is an allelic variant of a polynucleotide of (a)-(g) above, and
(k) a polynucleotide which encodes a species homologue of the protein of (h) or (l) above.
12 A composition comprising a protein, wherein said protein comprises an amino acid sequence selected from the group consisting of
(a) the amino acid sequence of SEQ ID NO 7,
(b) the ammo acid sequence of SEQ ID NO 7 from ammo acid 31 to amino acid 50,
(c) fragments of the ammo acid sequence of SEQ ID NO 7, and
(d) the ammo acid sequence encoded by the cDNA insert of clone AM610 deposited under accession number ATCC 98026, the protein being substantially free from other mammalian proteins
13 A composition comprising an isolated protein encoded by a 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 1 to nucleotide 483,
(c) a polynucleotide comprising the nucleotide sequence of the full length protein coding sequence of clone AM340 deposited under accession number ATCC 98026 ,
(d) a polynucleotide encoding the full length protem encoded by the cDNA insert of clone AM340 deposited under accession number ATCC 98026 ,
(e) a polynucleotide comprising the nucleotide sequence of the mature protein coding sequence of clone AM340 deposited under accession number ATCC 98026 ,
(f) a polynucleotide encoding the mature protein encoded by the cDNA insert of clone AM340 deposited under accession number ATCC 98026 ,
(g) a polynucleotide encoding a protein compπsing the amino acid sequence of SEQ ID NO 10,
(h) a polynucleotide encoding a protein comprising a fragment of the ammo acid sequence of SEQ ID NO 10 having biological activity,
(I) a polynucleotide which is an allelic variant of a polynucleotide of (a)-(f) above, and
(j) a polynucleotide which encodes a species homologue of the protein of
(g) or (h) above
14 A composition compπsing a protein, wherein said protein comprises an amino acid sequence selected from the group consisting of
(a) the amino acid sequence of SEQ ID NO 10,
(b) the amino acid sequence of SEQ ID NO 10 from amino acid 124 to
Figure imgf000113_0001
(c) fragments of the amino acid sequence of SEQ ID NO 10, and
(d) the amino acid sequence encoded by the cDNA insert of clone AM340 deposited under accession number ATCC 98026, the protein being substantially free from other mammalian proteins
15 A composition comprising an isolated protem encoded by a 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 1 1 from nucleotide 15 to nucleotide 462,
(c) a polynucleotide compπsing the nucleotide sequence of SEQ ID NO 1 1 from nucleotide 87 to nucleotide 462,
(d) a polynucleotide comprising the nucleotide sequence of the full length protein coding sequence of clone AM282 deposited under accession number ATCC 98026 ,
(e) a polynucleotide encoding the full length protem encoded by the cDNA insert of clone AM282 deposited under accession number ATCC 98026 ,
(0 a polynucleotide comprising the nucleotide sequence of the mature protein coding sequence of clone AM282 deposited under accession number ATCC 98026 ,
(g) a polynucleotide encoding the mature protein encoded by the cDNA insert of clone AM282 deposited under accession number ATCC 98026 ,
(h) a polynucleotide encoding a protein comprising the amino acid sequence of SEQ ID NO 12,
(0 a polynucleotide encoding a protein comprising a fragment of the amino acid sequence of SEQ ID NO 12 having biological activity,
(j) a polynucleotide which is an allelic vaπant of a polynucleotide of (a)-(g) above, and
(k) a polynucleotide which encodes a species homologue of the protein of (h) or (I) above
16 A composition comprising a protein, wherem said protein comprises an amino acid sequence selected from the group consisting of
(a) the ammo acid sequence of SEQ ID NO 12,
(b) the ammo acid sequence of SEQ ID NO 12 from amino acid 28 to ammo acid 47,
(c) fragments of the amino acid sequence of SEQ ID NO 12, and
(d) the amino acid sequence encoded by the cDNA insert of clone AM282 deposited under accession number ATCC 98026, the protein being substantially free from other mammalian proteins
17 A composition comprising an isolated protein encoded by a polynucleotide selected from the group consisting of
(a) a polynucleotide comprising the nucleotide sequence of SEQ ID NO 14,
(b) a polynucleotide comprising the nucleotide sequence of SEQ ID NO 14 from nucleotide 185 to nucleotide 519,
(c) a polynucleotide comprising the nucleotide sequence of SEQ ID NO 14 from nucleotide 260 to nucleotide 519,
(d) a polynucleotide comprising the nucleotide sequence of the full length protein coding sequence of clone AK647 deposited under accession number ATCC 98026 ,
(e) a polynucleotide encoding the full length protem encoded by the cDNA insert of clone AK647 deposited under accession number ATCC 98026 ,
(0 a polynucleotide comprising the nucleotide sequence of the mature protein coding sequence of clone AK647 deposited under accession number ATCC 98026 ,
(g) a polynucleotide encoding the mature protem encoded by the cDNA insert of clone AK647 deposited under accession number ATCC 98026 ,
(h) a polynucleotide encoding a protein comprising the amino acid sequence of SEQ ID NO 15,
(l) a polynucleotide encoding a protein comprising a fragment of the amino acid sequence of SEQ ID NO 15 having biological activity,
(j) a polynucleotide which is an allelic vaπant of a polynucleotide of (a)-(g) above, and
(k) a polynucleotide which encodes a species homologue of the protein of (h) or (l) above
18 A composition compπsing a protein, wherein said protein comprises an amino acid sequence selected from the group consisting of
(a) the amino acid sequence of SEQ ID NO 15,
(b) the ammo acid sequence of SEQ ID NO 15 from ammo acid 27 to amino acid 46,
(c) fragments of the amino acid sequence of SEQ ID NO 15, and
(d) the amino acid sequence encoded by the cDNA insert of clone AK647 deposited under accession number ATCC 98026, the protein being substantially free from other mammalian proteins
19 A composition comprising an isolated protein encoded by a polynucleotide selected from the group consisting of
(a) a polynucleotide comprising the nucleotide sequence of SEQ ID NO 17,
(b) a polynucleotide comprising the nucleotide sequence of SEQ ID NO 17 from nucleotide 257 to nucleotide 536,
(c) a polynucleotide comprising the nucleotide sequence of SEQ ID NO 17 from nucleotide 329 to nucleotide 536,
(d) a polynucleotide compπsing the nucleotide sequence of the full length protein coding sequence of clone AK583 deposited under accession number ATCC 98026 ,
(e) a polynucleotide encoding the full length protein encoded by the cDNA insert of clone AK583 deposited under accession number ATCC 98026 ,
(f) a polynucleotide comprising the nucleotide sequence of the mature protein coding sequence of clone AK583 deposited under accession number ATCC 98026 ,
(g) a polynucleotide encoding the mature protein encoded by the cDNA insert of clone AK583 deposited under accession number ATCC 98026 ,
(h) a polynucleotide encoding a protein comprising the amino acid sequence of SEQ ID NO 18,
(i) a polynucleotide encoding a protein compπsing a fragment of the amino acid sequence of SEQ ID NO 18 having biological activity,
(j) a polynucleotide which is an allelic variant of a polynucleotide of (a)-(g) above, and
(k) a polynucleotide which encodes a species homologue of the protein of (h) or (I) above
20 A composition comprising a protein, wherein said protein comprises an amino acid sequence selected from the group consisting of
(a) the amino acid sequence of SEQ ID NO 18,
(b) the ammo acid sequence of SEQ ID NO 18 from amino acid 14 to ammo acid 33,
(c) fragments of the amino acid sequence of SEQ ID NO 18, and (d) the ammo acid sequence encoded by the cDNA insert of clone AK583 deposited under accession number ATCC 98026, the protein being substantially free from other mammalian proteins
21 A composition comprising an isolated protein encoded by a polynucleotide selected from the group consisting of
(a) a polynucleotide comprising the nucleotide sequence of SEQ ID NO 20,
(b) a polynucleotide comprising the nucleotide sequence of SEQ ID NO 20 from nucleotide 179 to nucleotide 476,
(c) a polynucleotide comprising the nucleotide sequence of the full length protein coding sequence of clone AK533 deposited under accession number ATCC 98026 ,
(d) a polynucleotide encoding the full length protein encoded by the cDNA insert of clone AK533 deposited under accession number ATCC 98026 ,
(e) a polynucleotide comprising the nucleotide sequence of the mature protein coding sequence of clone AK533 deposited under accession number ATCC 98026 ,
(f) a polynucleotide encoding the mature protein encoded by the cDNA insert of clone AK533 deposited under accession number ATCC 98026 ,
(g) a polynucleotide encoding a protein comprising the ammo acid sequence of SEQ ID NO.21 ,
(h) a polynucleotide encoding a protein comprising a fragment of the amino acid sequence of SEQ ID NO 21 having biological activity,
(0 a polynucleotide which is an allelic variant of a polynucleotide of (a)-(f) above, and
(j) a polynucleotide which encodes a species homologue of the protein of (g) or (h) above
22 A composition comprising a protein, wherein said protein comprises an ammo acid sequence selected from the group consisting of
(a) the ammo acid sequence of SEQ ID NO 21 ,
(b) the amino acid sequence of SEQ ID NO 21 from ammo acid 35 to ammo acid 57,
(c) fragments of the amino acid sequence of SEQ ID NO 21, and (d) the ammo acid sequence encoded by the cDNA insert of clone AK533 deposited under accession number ATCC 98026, the protein being substantially free from other mammalian proteins
23 A composition comprising an isolated protem encoded by a polynucleotide selected from the group consisting of
(a) a polynucleotide comprising the nucleotide sequence of SEQ ID NO 23,
(b) a polynucleotide comprising the nucleotide sequence of SEQ ID NO 23 from nucleotide 220 to nucleotide 612,
(c) a polynucleotide comprising the nucleotide sequence of SEQ ID NO 23 from nucleotide 328 to nucleotide 612,
(d) a polynucleotide comprising the nucleotide sequence of the full length protein coding sequence of clone AK296 deposited under accession number ATCC 98026 ,
(e) a polynucleotide encoding the full length protem encoded by the cDNA insert of clone AK296 deposited under accession number ATCC 98026 ,
(f) a polynucleotide comprising the nucleotide sequence of the mature protein coding sequence of clone AK296 deposited under accession number ATCC 98026 ,
(g) a polynucleotide encoding the mature protein encoded by the cDNA insert of clone AK296 deposited under accession number ATCC 98026 ,
(h) a polynucleotide encoding a protein comprising the amino acid sequence of SEQ ID NO 24,
(l) a polynucleotide encoding a protem comprising a fragment of the ammo acid sequence of SEQ ID NO 24 having biological activity,
(j) a polynucleotide which is an allelic variant of a polynucleotide of (a)-(g) above, and
(k) a polynucleotide which encodes a species homologue of the protein of (h) or (I) above
24 A composition comprising a protein, wherein said protein comprises an ammo acid sequence selected from the group consisting of
(a) the amino acid sequence of SEQ ID NO 24, (b) the amino acid sequence of SEQ ID NO: 24 from amino acid 81 to amino acid 90;
(c) fragments of the amino acid sequence of SEQ ID NO:24; and
(d) the amino acid sequence encoded by the cDNA insert of clone AK296 deposited under accession number ATCC 98026; the protein being substantially free from other mammalian proteins.
25. A composition comprising an isolated protein encoded by a polynucleotide selected from the group consisting of:
(a) a polynucleotide comprising the nucleotide sequence of SEQ ID NO:26;
(b) a polynucleotide comprising the nucleotide sequence of SEQ ID NO:26 from nucleotide 58 to nucleotide 655;
(c) a polynucleotide comprising the nucleotide sequence of the full length protein coding sequence of clone H617 deposited under accession number ATCC 98026
(d) a polynucleotide encoding the full length protein encoded by the cDNA insert of clone H617 deposited under accession number ATCC 98026 ;
(e) a polynucleotide comprising the nucleotide sequence of the mature protein coding sequence of clone H617 deposited under accession number ATCC 98026
(f) a polynucleotide encoding the mature protein encoded by the cDNA insert of clone H617 deposited under accession number ATCC 98026 ;
(g) a polynucleotide encoding a protein comprising the amino acid sequence of SEQ ID NO:27;
(h) a polynucleotide encoding a protein comprising a fragment of the amino acid sequence of SEQ ID NO: 27 having biological activity;
(i) a polynucleotide which is an allelic variant of a polynucleotide of (a)-(f) above; and
(j) a polynucleotide which encodes a species homologue of the protein of
(g) or (h) above.
26. A composition comprising a protein, wherein said protein comprises an amino acid sequence selected from the group consisting of:
(a) the amino acid sequence of SEQ ID NO:27; (b) the amino acid sequence of SEQ ID NO 27 from amino acid 65 to amino
(c) fragments of the amino acid sequence of SEQ ID NO 27, and
(d) the ammo acid sequence encoded by the cDNA insert of clone H617 deposited under accession number ATCC 98026, the protein being substantially free from other mammalian proteins
27 A composition comprising an isolated protein encoded by a polynucleotide selected from the group consisting of
(a) a polynucleotide comprising the nucleotide sequence of SEQ ID NO 29,
(b) a polynucleotide comprising the nucleotide sequence of SEQ ID NO 29 from nucleotide 14 to nucleotide 391 ,
(c) a polynucleotide comprising the nucleotide sequence of the full length protein coding sequence of clone BB9 deposited under accession number ATCC 98026 ,
(d) a polynucleotide encoding the full length protein encoded by the cDNA insert of clone BB9 deposited under accession number ATCC 98026 ,
(e) a polynucleotide comprising the nucleotide sequence of the mature protein coding sequence of clone BB9 deposited under accession number ATCC 98026 ,
(f) a polynucleotide encoding the mature protein encoded by the cDNA insert of clone BB9 deposited under accession number ATCC 98026 ,
(g) a polynucleotide encoding a protein comprising the ammo acid sequence of SEQ ID NO 30,
(h) a polynucleotide encoding a protein comprising a fragment of the amino acid sequence of SEQ ID NO 30 having biological activity,
(0 a polynucleotide which is an allelic variant of a polynucleotide of (a)-(f) above, and
(j) a polynucleotide which encodes a species homologue of the protein of (g) or (h) above
28 A composition comprising a protein, whe aid protein comprises an amino acid sequence selected from the group consisting of
(a) the amino acid sequence of SEQ ID NO 30,
(b) the amino acid sequence of SEQ ID NO 30 from ammo acid 75 to ammo acid 94, (c) fragments of the amino acid sequence of SEQ ID NO:30; and
(d) the amino acid sequence encoded by the cDNA insert of clone BB9 deposited under accession number ATCC 98026; the protein being substantially free from other mammalian proteins.
29. A composition comprising an isolated protein encoded by a polynucleotide selected from the group consisting of:
(a) a polynucleotide comprising the nucleotide sequence of SEQ ID NO:32;
(b) a polynucleotide comprising the nucleotide sequence of SEQ ID NO:32 from nucleotide 61 to nucleotide 514;
(c) a polynucleotide comprising the nucleotide sequence of SEQ ID NO:32 from nucleotide 115 to nucleotide 514;
(d) a polynucleotide comprising the nucleotide sequence of the full length protein coding sequence of clone AW191 deposited under accession number ATCC 98026 ;
(e) a polynucleotide encoding the full length protein encoded by the cDNA insert of clone AW19I deposited under accession number ATCC 98026 ;
(f) a polynucleotide comprising the nucleotide sequence of the mature protein coding sequence of clone AW191 deposited under accession number ATCC 98026 ;
(g) a polynucleotide encoding the mature protein encoded by the cDNA insert of clone AW191 deposited under accession number ATCC 98026 ;
(h) a polynucleotide encoding a protein comprising the amino acid sequence of SEQ ID NO:33;
(i) a polynucleotide encoding a protein comprising a fragment of the amino acid sequence of SEQ ID NO:33 having biological activity;
(j) a polynucleotide which is an allelic variant of a polynucleotide of (a)-(g) above; and
(k) a polynucleotide which encodes a species homologue of the protein of (h) or (i) above.
30. A composition comprising a protein, wherein said protein comprises an amino acid sequence selected from the group consisting of:
(a) the amino acid sequence of SEQ ID NO: 33; (b) the ammo acid sequence of SEQ ID NO.33 from amino acid 24 to ammo
(c) fragments of the amino acid sequence of SEQ ID NO 33, and
(d) the ammo acid sequence encoded by the cDNA insert of clone AW 191 deposited under accession number ATCC 98026, the protein being substantially free from other mammalian proteins
31 A composition comprising an isolated protein encoded by a polynucleotide selected from the group consisting of.
(a) a polynucleotide compπsing the nucleotide sequence of SEQ ID NO.35,
(b) a polynucleotide comprising the nucleotide sequence of SEQ ID NO.35 from nucleotide 180 to nucleotide 525;
(c) a polynucleotide comprising the nucleotide sequence of SEQ ID NO 35 from nucleotide 339 to nucleotide 525,
(d) a polynucleotide comprising the nucleotide sequence of the full length protein coding sequence of clone AT211 deposited under accession number ATCC 98026 ,
(e) a polynucleotide encoding the full length protein encoded by the cDNA insert of clone AT211 deposited under accession number ATCC 98026 ;
(f) a polynucleotide comprising the nucleotide sequence of the mature protein coding sequence of clone AT21 1 deposited under accession number ATCC 98026 ,
(g) a polynucleotide encoding the mature protein encoded by the cDNA insert of clone AT21 1 deposited under accession number ATCC 98026 ,
(h) a polynucleotide encoding a protem comprising the amino acid sequence of SEQ ID NO.36,
(i) a polynucleotide encoding a protein compπsing a fragment of the ammo acid sequence of SEQ ID NO.36 having biological activity,
(j) a polynucleotide which is an allelic variant of a polynucleotide of (a)-(g) above; and
(k) a polynucleotide which encodes a species homologue of the protein of (h) or (i) above.
32 A composition comprising a protein, wherein said protein comprises an amino acid sequence selected from the group consisting of
(a) the ammo acid sequence of SEQ ID NO 36,
(b) the amino acid sequence of SEQ ID NO 36 from ammo acid 1 to amino
(c) fragments of the amino acid sequence of SEQ ID NO 36, and
(d) the ammo acid sequence encoded by the cDNA insert of clone AT211 deposited under accession number ATCC 98026, the protein being substantially free from other mammalian proteins
33 A composition comprising an isolated protein encoded by a polynucleotide selected from the group consisting of
(a) a polynucleotide compπsing the nucleotide sequence of SEQ ID NO.38,
(b) a polynucleotide comprising the nucleotide sequence of SEQ ID NO 38 from nucleotide 225 to nucleotide 677,
(c) a polynucleotide comprising the nucleotide sequence of SEQ ID NO 38 from nucleotide 390 to nucleotide 677,
(d) a polynucleotide comprising the nucleotide sequence of the full length protein coding sequence of clone AT205 deposited under accession number ATCC 98026 ,
(e) a polynucleotide encoding the full length protein encoded by the cDNA insert of clone AT205 deposited under accession number ATCC 98026 ,
(0 a polynucleotide comprising the nucleotide sequence of the mature protem coding sequence of clone AT205 deposited under accession number ATCC 98026 ,
(g) a polynucleotide encoding the mature protein encoded by the cDNA insert of clone AT205 deposited under accession number ATCC 98026 ;
(h) a polynucleotide encoding a protein comprising the ammo acid sequence of SEQ ID NO:39,
(i) a polynucleotide encoding a protein comprising a fragment of the ammo acid sequence of SEQ ID NO 39 having biological activity,
(j) a polynucleotide which is an allelic variant of a polynucleotide of (a)-(g) above; and (k) a polynucleotide which encodes a species homologue of the protein of (h) or (i) above.
34. A composition comprising a protein, wherein said protein comprises an amino acid sequence selected from the group consisting of:
(a) the amino acid sequence of SEQ ID NO:39;
(b) the amino acid sequence of SEQ ID NO: 39 from amino acid 6 to amino acid 25;
(c) fragments of the amino acid sequence of SEQ ID NO: 39; and
(d) the amino acid sequence encoded by the cDNA insert of clone AT205 deposited under accession number ATCC 98026; the protein being substantially free from other mammalian proteins.
35. A composition comprising an isolated protein encoded by a polynucleotide selected from the group consisting of:
(a) a polynucleotide comprising the nucleotide sequence of SEQ ID NO:40;
(b) a polynucleotide comprising the nucleotide sequence of SEQ ID NO:40 from nucleotide 128 to nucleotide 508;
(c) a polynucleotide comprising the nucleotide sequence of SEQ ID NO:40 from nucleotide 200 to nucleotide 508;
(d) a polynucleotide comprising the nucleotide sequence of the full length protein coding sequence of clone AS34 deposited under accession number ATCC 98026
(e) a polynucleotide encoding the full length protein encoded by the cDNA insert of clone AS34 deposited under accession number ATCC 98026 ;
(0 a polynucleotide comprising the nucleotide sequence of the mature protein coding sequence of clone AS34 deposited under accession number ATCC 98026
(g) a polynucleotide encoding the mature protein encoded by the cDNA insert of clone AS34 deposited under accession number ATCC 98026 ;
(h) a polynucleotide encoding a protein comprising the amino acid sequence of SEQ ID NO:41 ;
(i) a polynucleotide encoding a protein comprising a fragment of the amino acid sequence of SEQ ID NO:41 having biological activity; (j) a polynucleotide which is an allelic variant of a polynucleotide of (a)-(g) above, and
(k) a polynucleotide which encodes a species homologue of the protein of (h) or 0) above
36 A composition comprising a protein, wherein said protein compπses an amino acid sequence selected from the group consisting of
(a) the ammo acid sequence of SEQ ID NO 41 ,
(b) the amino acid sequence of SEQ ID NO 41 from ammo acid 27 to amino acid 46,
(c) fragments of the amino acid sequence of SEQ ID NO 41 , and
(d) the ammo acid sequence encoded by the cDNA insert of clone AS34 deposited under accession number ATCC 98026, the protein being substantially tree from other mammalian proteins
37 A composition comprising an isolated protem encoded by a polynucleotide selected from the group consisting of
(a) a polynucleotide comprising the nucleotide sequence of SEQ ID NO 43,
(b) a polynucleotide comprising the nucleotide sequence of SEQ ID NO 43 from nucleotide 23 to nucleotide 676,
(c) a polynucleotide comprising the nucleotide sequence of the full length protein coding sequence of clone AS32 deposited under accession number ATCC 98026
(d) a polynucleotide encoding the full length protein encoded by the cDNA insert of clone AS32 deposited under accession number ATCC 98026 ,
(e) a polynucleotide comprising the nucleotide sequence of the mature protein coding sequence of clone AS32 deposited under accession number ATCC 98026
»
(f) a polynucleotide encoding the mature protein encoded by the cDNA insert of clone AS32 deposited under accession number ATCC 98026 ,
(g) a polynucleotide encoding a protein compπsing the amino acid sequence of SEQ ID NO 44,
(h) a polynucleotide encoding a protein comprising a fragment of the amino acid sequence of SEQ ID NO 44 having biological activity, (i) a polynucleotide which is an allelic variant of a polynucleotide of (a)-(f) above; and
(j) a polynucleotide which encodes a species homologue of the protein of (g) or (h) above.
38. A composition comprising a protein, wherein said protein comprises an amino acid sequence selected from the group consisting of:
(a) the amino acid sequence of SEQ ID NO:44;
(b) the amino acid sequence of SEQ ID NO:44 from amino acid 78 to amino acid 97;
(c) fragments of the amino acid sequence of SEQ ID NO:44; and
(d) the amino acid sequence encoded by the cDNA insert of clone AS32 deposited under accession number ATCC 98026; the protein being substantially free from other mammalian proteins.
39. A composition comprising an isolated protein encoded by a polynucleotide selected from the group consisting of:
(a) a polynucleotide comprising the nucleotide sequence of SEQ ID NO:46;
(b) a polynucleotide comprising the nucleotide sequence of SEQ ID NO:46 from nucleotide 132 to nucleotide 479;
(c) a polynucleotide comprising the nucleotide sequence of SEQ ID NO:46 from nucleotide 201 to nucleotide 479;
(d) a polynucleotide comprising the nucleotide sequence of the full length protein coding sequence of clone AR260 deposited under accession number ATCC 98026 ;
(e) a polynucleotide encoding the full length protein encoded by the cDNA insert of clone AR260 deposited under accession number ATCC 98026 ;
(0 a polynucleotide comprising the nucleotide sequence of the mature protein coding sequence of clone AR260 deposited under accession number ATCC 98026 ;
(g) a polynucleotide encoding the mature protein encoded by the cDNA insert of clone AR260 deposited under accession number ATCC 98026 ;
(h) a polynucleotide encoding a protein comprising the amino acid sequence of SEQ ID NO:47; (1) a polynucleotide encoding a protein comprising a fragment of the ammo acid sequence of SEQ ID NO 47 having biological activity,
(j) a polynucleotide which is an allelic vaπant of a polynucleotide of (a)-(g) above, and
(k) a polynucleotide which encodes a species homologue of the protein of (h) or (1) above
40 A composition comprising a protein, wherein said protein compπses an ammo acid sequence selected from the group consisting of
(a) the ammo acid sequence of SEQ ID NO 47,
(b) the amino acid sequence of SEQ ID NO 47 from ammo acid 40 to amino acid 59,
(c) fragments of the amino acid sequence of SEQ ID NO 47, and
(d) the amino acid sequence encoded by the cDNA insert of clone AR260 deposited under accession number ATCC 98026, the protein being substantially free from other mammalian proteins
41 A composition comprising an isolated protein encoded by a polynucleotide selected from the group consisting of
(a) a polynucleotide comprising the nucleotide sequence of SEQ ID NO 50,
(b) a polynucleotide comprising the nucleotide sequence of SEQ ID NO 50 from nucleotide 1 to nucleotide 332,
(c) a polynucleotide comprising the nucleotide sequence of the full length protein coding sequence of clone K640 deposited under accession number ATCC 98026
(d) a polynucleotide encoding the full length protein encoded by the cDNA insert of clone K640 deposited under accession number ATCC 98026 ,
(e) a polynucleotide comprising the nucleotide sequence of the mature protein coding sequence of clone K640 deposited under accession number ATCC 98026
(f) a polynucleotide encoding the mature protem encoded by the cDNA insert of clone K640 deposited under accession number ATCC 98026 ,
(g) a polynucleotide encoding a protem comprising the ammo acid sequence of SEQ ID NO:51; (h) a polynucleotide encoding a protein comprising a fragment of the amino acid sequence of SEQ ID NO:51 having biological activity;
(i) a polynucleotide which is an allelic variant of a polynucleotide of (a)-(f) above; and
(j) a polynucleotide which encodes a species homologue of the protein of (g) or (h) above.
42. A composition comprising a protein, wherein said protein comprises an amino acid sequence selected from the group consisting of:
(a) the amino acid sequence of SEQ ID NO:51 ;
(b) the amino acid sequence of SEQ ID NO:51 from amino acid 11 to amino acid 30;
(c) fragments of the amino acid sequence of SEQ ID NO:51 ; and
(d) the amino acid sequence encoded by the cDNA insert of clone K640 deposited under accession number ATCC 98026; the protein being substantially free from other mammalian proteins.
43. A composition comprising an isolated protein encoded by a polynucleotide selected from the group consisting of:
(a) a polynucleotide comprising the nucleotide sequence of SEQ ID NO:54;
(b) a polynucleotide comprising the nucleotide sequence of SEQ ID NO: 54 from nucleotide 71 to nucleotide 377;
(c) a polynucleotide comprising the nucleotide sequence of the full length protein coding sequence of clone K39 deposited under accession number ATCC 98026 ;
(d) a polynucleotide encoding the full length protein encoded by the cDNA insert of clone K39 deposited under accession number ATCC 98026 ;
(e) a polynucleotide comprising the nucleotide sequence of the mature protein coding sequence of clone K39 deposited under accession number ATCC 98026 ;
(0 a polynucleotide encoding the mature protein encoded by the cDNA insert of clone K39 deposited under accession number ATCC 98026 ;
(g) a polynucleotide encoding a protein comprising the amino acid sequence of SEQ ID NO:55;
(h) a polynucleotide encoding a protein comprising a fragment of the amino acid sequence of SEQ ID NO:55 having biological activity; (i) a polynucleotide which is an allelic variant of a polynucleotide of (a)-(f) above, and
(j) a polynucleotide which encodes a species homologue of the protein of (g) or (h) above
44 A composition comprising a protein, wherein said protein comprises an amino acid sequence selected from the group consisting of
(a) the ammo acid sequence of SEQ ID NO 55,
(b) the amino acid sequence of SEQ ID NO 55 from amino acid 62 to amino ,
(c) fragments of the amino acid sequence of SEQ ID NO 55, and
(d) the ammo acid sequence encoded by the cDNA insert of clone K39 deposited under accession number ATCC 98026, the protein being substantially free from other mammalian proteins
45 A composition comprising an isolated protein encoded by a polynucleotide selected from the group consisting of
(a) a polynucleotide comprising the nucleotide sequence of SEQ ID NO 57,
(b) a polynucleotide compπsing the nucleotide sequence of SEQ ID NO 57 from nucleotide 194 to nucleotide 423,
(c) a polynucleotide comprising the nucleotide sequence of the full length protein coding sequence of clone AT319 deposited under accession number ATCC 98026 ,
(d) a polynucleotide encoding the full length protein encoded by the cDNA insert of clone AT319 deposited under accession number ATCC 98026 ,
(e) a polynucleotide comprising the nucleotide sequence of the mature protein coding sequence of clone AT319 deposited under accession number ATCC 98026 ,
(f) a polynucleotide encoding the mature protein encoded by the cDNA insert of clone AT319 deposited under accession number ATCC 98026 ,
(g) a polynucleotide encoding a protein comprising the amino acid sequence of SEQ ID NO 58,
(h) a polynucleotide encoding a protein comprising a fragment of the ammo acid sequence of SEQ ID NO 58 having biological activity. (1) a polynucleotide which is an allelic vaπant of a polynucleotide of (a)-(f) above, and
(j) a polynucleotide which encodes a species homologue of the protein of
(g) or (h) above
46 A composition comprising a protein, wherein said protein comprises an amino acid sequence selected from the group consisting of
(a) the ammo acid sequence of SEQ ID NO 58,
(b) the amino acid sequence of SEQ ID NO 58 from amino acid 2 to amino acid 21 ,
(c) fragments of the ammo acid sequence of SEQ ID NO 58, and
(d) the amino acid sequence encoded by the cDNA insert of clone AT319 deposited under accession number ATCC 98026, the protein being substantially free from other mammalian proteins
PCT/US1997/006139 1996-04-18 1997-04-14 Secreted proteins WO1997039123A2 (en)

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JP53727097A JP2001509004A (en) 1996-04-18 1997-04-14 Secreted protein

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US08/634,325 1996-04-18

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Cited By (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1999000408A2 (en) * 1997-05-13 1999-01-07 Incyte Pharmaceuticals, Inc. Human transmembrane protein from the transmembrane 4 superfamily
WO1999000410A2 (en) * 1997-06-27 1999-01-07 Incyte Pharmaceuticals, Inc. Human extracellular matrix proteins
WO1999055864A1 (en) * 1998-04-28 1999-11-04 Ono Pharmaceutical Co., Ltd. NOVEL POLYPEPTIDE, cDNA ENCODING THE SAME AND UTILIZATION THEREOF
WO2000037630A1 (en) * 1998-12-23 2000-06-29 Genetics Institute, Inc. Secreted proteins
EP1037898A1 (en) * 1997-06-27 2000-09-27 Genetics Institute, Inc. Secreted proteins
WO2001094587A2 (en) * 2000-06-06 2001-12-13 Incyte Genomics, Inc. Extracellular messengers

Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1996000738A1 (en) * 1994-06-30 1996-01-11 Warner-Lambert Company Endothelin antagonists ii

Patent Citations (1)

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Publication number Priority date Publication date Assignee Title
WO1996000738A1 (en) * 1994-06-30 1996-01-11 Warner-Lambert Company Endothelin antagonists ii

Non-Patent Citations (2)

* Cited by examiner, † Cited by third party
Title
DATABASE EMBL EST Sequence HS096209 Acc. No. H62096 yu40d08.r1 Homo sapiens cDNA clone 236271 5', 8 October 1995 XP002042762 cited in the application & L. HILLIER ET AL.: "The WashU-Merck EST Project" *
JOURNAL OF CELLULAR BIOCHEMISTRY - SUPPLEMENT, vol. 21A, 10 March 1995, page 19 XP002027246 K JACOBS ET AL: "A novel method for isolating eukaryotic cDNA clones encoding secreted proteins" *

Cited By (11)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1999000408A2 (en) * 1997-05-13 1999-01-07 Incyte Pharmaceuticals, Inc. Human transmembrane protein from the transmembrane 4 superfamily
WO1999000408A3 (en) * 1997-05-13 1999-04-15 Incyte Pharma Inc Human transmembrane protein from the transmembrane 4 superfamily
WO1999000410A2 (en) * 1997-06-27 1999-01-07 Incyte Pharmaceuticals, Inc. Human extracellular matrix proteins
WO1999000410A3 (en) * 1997-06-27 1999-03-18 Incyte Pharma Inc Human extracellular matrix proteins
EP1037898A1 (en) * 1997-06-27 2000-09-27 Genetics Institute, Inc. Secreted proteins
US6303765B1 (en) 1997-06-27 2001-10-16 Incyte Genomics, Inc. Human extracellular matrix proteins
EP1037898A4 (en) * 1997-06-27 2003-04-16 Inst Genetics Llc Secreted proteins
WO1999055864A1 (en) * 1998-04-28 1999-11-04 Ono Pharmaceutical Co., Ltd. NOVEL POLYPEPTIDE, cDNA ENCODING THE SAME AND UTILIZATION THEREOF
WO2000037630A1 (en) * 1998-12-23 2000-06-29 Genetics Institute, Inc. Secreted proteins
WO2001094587A2 (en) * 2000-06-06 2001-12-13 Incyte Genomics, Inc. Extracellular messengers
WO2001094587A3 (en) * 2000-06-06 2002-12-27 Incyte Genomics Inc Extracellular messengers

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AU2801697A (en) 1997-11-07
WO1997039123A3 (en) 1998-02-26
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JP2001509004A (en) 2001-07-10

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