Fibroblast Growth Factor-13

Abstract

The present invention relates to a novel FGF-13 protein which is a member of the fibroblast growth factor (FGF) family. In particular, isolated nucleic acid molecules are provided encoding the human FGF-13 protein. FGF-13 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 FGF-13 activity. Also provided are diagnostic methods for detecting FGF-13-related disorders and therapeutic methods for treating FGF-13-related disorders. Disclosed is a method of prolonging dopaminergic neuron survival.

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 FGF-13 polypeptide having the amino acid sequence at positions −22 to 193 of SEQ ID NO:2;    (b) a nucleotide sequence encoding the mature FGF-13 polypeptide having the amino acid sequence from about position 1 to about position 193 in SEQ ID NO:2;    (c) a nucleotide sequence encoding the FGF-13 polypeptide having the complete amino acid sequence encoded by the cDNA clone contained in ATCC™ Deposit No. 97147;    (d) a nucleotide sequence encoding the mature FGF-13 polypeptide having the amino acid sequence encoded by the cDNA clone contained in ATCC™ Deposit No. 97147; and    (e) a nucleotide sequence complementary to any of the nucleotide sequences in (a), (b), (c) or (d) above.    
   
   
       2 . The nucleic acid molecule of  claim 1  wherein said polynucleotide has the complete nucleotide sequence in FIGS.  1 A-D (SEQ ID NO:1).  
   
   
       3 . The nucleic acid molecule of  claim 1  wherein said polynucleotide has the nucleotide sequence in FIGS.  1 A-D (SEQ ID NO:1) encoding the FGF-13 polypeptide having the complete amino acid sequence in SEQ ID NO:2.  
   
   
       4 . The nucleic acid molecule of  claim 1  wherein said polynucleotide has the nucleotide sequence in FIGS.  1 A-D (SEQ NO:1) encoding the mature FGF-13 polypeptide having the amino acid sequence from about 1 to about 193 in SEQ ID NO:2.  
   
   
       5 . 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-193 of SEQ ID NO:2, where n is an integer other than zero in the range of −23 to +10;    (b) a nucleotide sequence encoding a polypeptide comprising the amino acid sequence of residues −22 to -m of SEQ ID NO:2, where m is an integer in the range of 154 to 192;    (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 FGF-13 amino acid sequence encoded by the cDNA clone contained in ATCC™ Deposit No. 97147 wherein said portion excludes from 1 to about 33 amino acids from the amino terminus of said complete amino acid sequence encoded by the cDNA clone contained in ATCC™ Deposit No. 97147;    (e) a nucleotide sequence encoding a polypeptide consisting of a portion of the complete FGF-13 amino acid sequence encoded by the cDNA clone contained in ATCC™ Deposit No. 97147 wherein said portion excludes from 1 to about 39 amino acids from the carboxy terminus of said complete amino acid sequence encoded by the cDNA clone contained in ATCC™ Deposit No. 97147; and    (f) a nucleotide sequence encoding a polypeptide consisting of a portion of the complete FGF-13 amino acid sequence encoded by the cDNA clone contained in ATCC™ Deposit No. 97147 wherein said portion includes a combination of any of the amino terminal and carboxy terminal deletions in (d) and (e), above.    
   
   
       6 . The nucleic acid molecule of  claim 1  wherein said polynucleotide has the complete nucleotide sequence of the cDNA clone contained in, ATCC™ Deposit No. 97147.  
   
   
       7 . The nucleic acid molecule of  claim 1  wherein said polynucleotide has the nucleotide sequence encoding the FGF-13 polypeptide having the complete amino acid sequence encoded by the cDNA clone contained in ATCC™ Deposit No. 97147.  
   
   
       8 . The nucleic acid molecule of  claim 1  wherein said polynucleotide has the nucleotide sequence encoding the mature FGF-13 polypeptide having the amino acid sequence encoded by the cDNA clone contained in ATCC™ Deposit No. 97147.  
   
   
       9 . 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) or (d) 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.  
   
   
       10 . An isolated nucleic acid molecule comprising a polynucleotide which encodes the amino acid sequence of an epitope-bearing portion of an FGF-13 polypeptide having an amino acid sequence in (a), (b), (c) or (d) of  claim 1 .  
   
   
       11 . The isolated nucleic acid molecule of  claim 10 , which encodes an epitope-bearing portion of an FGF-13 polypeptide wherein the amino acid sequence of said portion is selected from the group of sequences in SEQ NO:2 consisting of: about Gln 22 to about Asn 32, about Asn 34 to about Met 43, about Gln 46 to about Tyr 55, about Ser 59 to about Val 66, about Val 68 to about Phe 83, about Val 88 to about Glu 105, about Met 110 to about Val 128, about Phe 153 to about His 173, about Leu 178 to about Gln 182, about Phe 185 to about Gln 194, and about Val 198 to about Gln 213.  
   
   
       12 . A method for making a recombinant vector comprising inserting an isolated nucleic acid molecule of  claim 1  into a vector.  
   
   
       13 . A recombinant vector produced by the method of  claim 12 .  
   
   
       14 . A method of making a recombinant host cell comprising introducing the recombinant vector of  claim 13  into a host cell.  
   
   
       15 . A recombinant host cell produced by the method of  claim 14 .  
   
   
       16 . A recombinant method for producing an FGF-13 polypeptide, comprising culturing the recombinant host cell of  claim 15  under conditions such that said polypeptide is expressed and recovering said polypeptide.  
   
   
       17 . An isolated FGF-13 polypeptide comprising an amino acid sequence at least 95% identical to a sequence selected from the group consisting of: 
 (a) the amino acid sequence at positions −22 to 193 of SEQ ID NO:2;    (b) the amino acid sequence from about position 1 to about position 193 in SEQ ID NO:2;    (c) the complete amino acid sequence encoded by the cDNA clone contained in ATCC™ Deposit No. 97147; and    (d) the mature FGF-13 amino acid sequence encoded by the cDNA clone contained in ATCC™ Deposit No. 97147.    
   
   
       18 . An isolated polypeptide comprising an epitope-bearing portion of the FGF-13 protein, wherein the amino acid sequence of said portion is selected from the group of sequences in SEQ ID NO:2 consisting of: about Gln 22 to about Asn 32, about Asn 34 to about Met 43, about Gln 46 to about Tyr 55, about Ser 59 to about Val 66, about Val 68 to about Phe 83, about Val 88 to about Glu 105, about Met 110 to about Val 128, about Phe 153 to about His 173, about Leu 178 to about Gln 182, about Phe 185 to about Gin 194, and about Val 198 to about Gln 213.  
   
   
       19 . An isolated antibody that binds specifically to an FGF-13 polypeptide of  claim 17 .  
   
   
       20 . 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 of a portion of the sequence shown in FIGS.  1 A-D (SEQ ID NO:1) wherein said portion comprises at least 50 contiguous nucleotides; and    (b) a nucleotide sequence complementary to any nucleotide sequence in (a) above.    
   
   
       21 . A method of increasing the survival of dopaminergic neurons comprising contacting a life prolonging concentration of FGF-13 with dopaminergic neurons.  
   
   
       22 . The method of  claim 22  wherein said dopaminergic neuron is in a Parkinson's Disease patient.