Cardiotrophin-like cytokine

Abstract

The present invention relates to a novel CLC protein which is a member of the IL-6 cytokine family. In particular, isolated nucleic acid molecules are provided encoding the human CLC protein. CLC polypeptides are also provided as are vectors, host cells and recombinant methods for producing the same. The invention further relates to screening methods for identifying agonists and antagonists of CLC activity. Also provided are diagnostic methods for detecting cardiac and immune system-related disorders and therapeutic methods for treating cardiac and immune system-related disorders.

What is claimed is:  
     
         1 . An isolated nucleic acid molecule nucleic acid molecule comprising a polynucleotide having a nucleotide sequence at least 95% identical to a sequence selected from the group consisting of: 
 (a) a nucleotide sequence encoding the CLC polypeptide having the complete amino acid sequence in SEQ ID NO:2 (i.e., positions −27 to 198 of SEQ ID NO:2);    (b) a nucleotide sequence encoding the CLC polypeptide having the complete amino acid sequence in SEQ ID NO:2 excepting the N-terminal methionine (i.e., positions −26 to 198 of SEQ ID NO:2);    (c) a nucleotide sequence encoding the predicted mature CLC polypeptide having the amino acid sequence at positions 1 to 198 in SEQ ID NO:2;    (d) a nucleotide sequence encoding the CLC polypeptide having the complete amino acid sequence encoded by the cDNA clone contained in ATCC Deposit No. 209122;    (e) a nucleotide sequence encoding the CLC polypeptide having the complete amino acid sequence excepting the N-terminal methionine encoded by the cDNA clone contained in ATCC Deposit No. 209122;    (f) a nucleotide sequence encoding the mature CLC polypeptide having the amino acid sequence encoded by the cDNA clone contained in ATCC Deposit No. 209122; and    (g) a nucleotide sequence complementary to any of the nucleotide sequences in (a), (b), (c), (d), (e) or (f) above.    
     
     
         2 . The nucleic acid molecule of  claim 1  wherein said polynucleotide has the complete nucleotide sequence in FIGS. 1A and 1B (SEQ ID NO:1).  
     
     
         3 . The nucleic acid molecule of  claim 1  wherein said polynucleotide has the nucleotide sequence in FIGS. 1A and 1B (SEQ ID NO:1) encoding the CLC polypeptide having the amino acid sequence in positions −27 to 198 of SEQ ID NO:2.  
     
     
         4 . The nucleic acid molecule of  claim 1  wherein said polynucleotide has the nucleotide sequence in FIGS. 1A and 1B (SEQ ID NO:1) encoding the CLC polypeptide having the amino acid sequence in positions −26 to 198 of SEQ ID NO:2.  
     
     
         5 . The nucleic acid molecule of  claim 1  wherein said polynucleotide has the nucleotide sequence in FIGS. 1A and 1B (SEQ ID NO:1) encoding the mature CLC polypeptide having the amino acid sequence from about 1 to about 198 in SEQ ID NO:2.  
     
     
         6 . An isolated nucleic acid molecule comprising a polynucleotide having a nucleotide sequence at least 95% identical to a sequence selected from the group consisting of: 
 (a) a nucleotide sequence encoding a polypeptide comprising the amino acid sequence of residues n-198 of SEQ ID NO:2, where n is an integer in the range of −27-10;    (b) a nucleotide sequence encoding a polypeptide comprising the amino acid sequence of residues −26-m of SEQ ID NO:2, where m is an integer in the range of 188 to 197;    (c) a nucleotide sequence encoding a polypeptide having the amino acid sequence consisting of residues n-m of SEQ ID NO:2, where n and m are integers as defined respectively in (a) and (b) above; and    (d) a nucleotide sequence encoding a polypeptide consisting of a portion of the complete CLC amino acid sequence encoded by the cDNA clone contained in ATCC Deposit No. 209122 wherein said portion excludes from 1 to about 36 amino acids from the amino terminus of said complete amino acid sequence;    (e) a nucleotide sequence encoding a polypeptide consisting of a portion of the complete CLC amino acid sequence encoded by the cDNA clone contained in ATCC Deposit No. 209122 wherein said portion excludes from 1 to about 10 amino acids from the carboxy terminus of said complete amino acid sequence; and    (f) a nucleotide sequence encoding a polypeptide consisting of a portion of the complete CLC amino acid sequence encoded by the cDNA clone contained in ATCC Deposit No. 209122 wherein said portion include a combination of any of the amino terminal and carboxy terminal deletions in (d) and (e), above.    
     
     
         7 . The nucleic acid molecule of  claim 1  wherein said polynucleotide has the complete nucleotide sequence of the cDNA clone contained in ATCC Deposit No. 209122.  
     
     
         8 . The nucleic acid molecule of  claim 1  wherein said polynucleotide has the nucleotide sequence encoding the CLC polypeptide having the complete amino acid sequence excepting the N-terminal methionine encoded by the cDNA clone contained in ATCC Deposit No. 209122.  
     
     
         9 . The nucleic acid molecule of  claim 1  wherein said polynucleotide has the nucleotide sequence encoding the mature CLC polypeptide having the amino acid sequence encoded by the cDNA clone contained in ATCC Deposit No. 209122.  
     
     
         10 . An isolated nucleic acid molecule comprising a polynucleotide which hybridizes under stringent hybridization conditions to a polynucleotide having a nucleotide sequence identical to a nucleotide sequence in (a), (b), (c), (d), (e), (f), or (g) of  claim 1  wherein said polynucleotide which hybridizes does not hybridize under stringent hybridization conditions to a polynucleotide having a nucleotide sequence consisting of only A residues or of only T residues.  
     
     
         11 . An isolated nucleic acid molecule comprising a polynucleotide which encodes the amino acid sequence of an epitope-bearing portion of a CLC polypeptide having an amino acid sequence in (a), (b), (c), (d), (e), or (f) of  claim 1 .  
     
     
         12 . The isolated nucleic acid molecule of  claim 10 , which encodes an epitope-bearing portion of a CLC polypeptide wherein the amino acid sequence of said portion is selected from the group of sequences in SEQ ID NO:2 consisting of: about Leu-1 to about Pro-9, from about Ser-12 to about Thr-20, from about Phe-42 to about Pro-50, from about Leu-52 to about Ala-60, from about Asn-71 to about Glu-92, from about Asn-94 to about Arg-104, from about Leu-135 to about Pro-144, from about His-148 to about Asp-157, and from about Asp-174 to about Gln-183.  
     
     
         13 . A method for making a recombinant vector comprising inserting an isolated nucleic acid molecule of  claim 1  into a vector.  
     
     
         14 . A recombinant vector produced by the method of  claim 13 .  
     
     
         15 . A method of making a recombinant host cell comprising introducing the recombinant vector of  claim 14  into a host cell.  
     
     
         16 . A recombinant host cell produced by the method of  claim 15 .  
     
     
         17 . A recombinant method for producing a CLC polypeptide, comprising culturing the recombinant host cell of  claim 16  under conditions such that said polypeptide is expressed and recovering said polypeptide.  
     
     
         18 . An isolated CLC polypeptide comprising an amino acid sequence at least 95% identical to a sequence selected from the group consisting of: 
 (a) the amino acid sequence of the full-length CLC polypeptide having the complete amino acid sequence shown in SEQ ID NO:2 (i.e., positions −27 to 198 of SEQ ID NO:2);    (b) the amino acid sequence of the full-length CLC polypeptide having the complete amino acid sequence shown in SEQ ID NO:2 excepting the N-terminal methionine (i.e., positions −26 to 198 of SEQ ID NO:2);    (c) the amino acid sequence of the mature CLC polypeptide having the complete amino acid sequence shown in SEQ ID NO:2 (i.e., positions 1 to 198 of SEQ ID NO:2);    (d) the amino acid sequence of the full-length CLC polypeptide encoded by the cDNA clone contained in ATCC Deposit No. 209122;    (e) the amino acid sequence of the full-length CLC polypeptide having the complete amino acid sequence, excepting the N-terminal methionine, encoded by the cDNA clone contained in ATCC Deposit No. 209122, and;    (f) the amino acid sequence of the mature CLC polypeptide encoded by the cDNA clone contained in ATCC Deposit No. 209122.    
     
     
         19 . An isolated polypeptide comprising an epitope-bearing portion of the CLC protein, wherein said portion is selected from the group consisting of: a polypeptide comprising amino acid residues from about Leu-1 to about Pro-9, from about Ser-12 to about Thr-20, from about Phe-42 to about Pro-50, from about Leu-52 to about Ala-60, from about Asn-71 to about Glu-92, from about Asn-94 to about Arg-104, from about Leu-135 to about Pro-144, from about His-148 to about Asp-157, and from about Asp-174 to about Gln-183.  
     
     
         20 . An isolated antibody that binds specifically to a CLC polypeptide of  claim 18 .  
     
     
         21 . An isolated nucleic acid molecule comprising a polynucleotide having a sequence at least 95% identical to a sequence selected from the group consisting of: 
 (a) the nucleotide sequence shown as SEQ ID NO:7;    (b) the nucleotide sequence of a portion of the sequence shown in FIGS. 1A and 1B (SEQ ID NO:1) wherein said portion comprises at least 50 contiguous nucleotides from nucleotide 400 to 900 and from 1375 to 1710;    (c) the nucleotide sequence of a portion of the sequence shown in FIGS. 1A and 1B (SEQ ID NO:1) wherein said portion consists of nucleotides 1-1000, 1-1250, 1-1500, 250-1000, 250-1250, 250-1500, 500-1000, 500-1250, 500-1500, 750-1000, 750-1250, 750-1500, 250-600, 400-600, 1000-1710, 1250-1710, 1000-1250, and 1000-1500; and    (d) a nucleotide sequence complementary to any of the nucleotide sequences in (a), (b), or (c) above.