Novel murine and human kinases

Abstract

The invention is directed to purified and isolated novel murine and human kinase polypeptides, the nucleic acids encoding such polypeptides, processes for production of recombinant forms of such polypeptides, antibodies generated against these polypeptides, fragmented peptides derived from these polypeptides, and the uses of the above.

1 . An isolated nucleic acid molecule selected from the group consisting of: 
 (a) the DNA sequence of SEQ ID NOs:1-7;    (b) an isolated nucleic acid molecule encoding an amino acid sequence comprising the sequence of SEQ ID NOs:8-14;    (c) an isolated nucleic acid molecule that hybridizes to either strand of a denatured, double-stranded DNA comprising the nucleic acid sequence of (a) or (b) under conditions of moderate stringency in 50% formamide and 6×SSC, at 42° C. with washing conditions of 60° C., 0.5×SSC, 0.1% SDS;    (d) an isolated nucleic acid molecule derived by in vitro mutagenesis from SEQ ID NOs:1-7;    (e) an isolated nucleic acid molecule degenerate from SEQ ID NOs:1-7 as a result of the genetic code; and    (f) an isolated nucleic acid molecule that is at least 80% identical to any of (a)-(e).    
     
     
         2 . A recombinant vector that directs the expression of the nucleic acid molecule of  claim 1 .  
     
     
         3 . An isolated polypeptide encoded by the nucleic acid molecule of  claim 1 .  
     
     
         4 . An isolated polypeptide according to  claim 3  in non-glycosylated form.  
     
     
         5 . Isolated antibodies that bind to a polypeptide of  claim 3 .  
     
     
         6 . Isolated antibodies according to  claim 5 , wherein the antibodies are monoclonal antibodies.  
     
     
         7 . A host cell transfected or transduced with the vector of  claim 2 .  
     
     
         8 . A method for the production of kinase polypeptide comprising culturing a host cell of  claim 2  under conditions promoting expression, and recovering the polypeptide from the culture medium.  
     
     
         9 . The method of  claim 8 , wherein the host cell is selected from the group consisting of bacterial cells, yeast cells, plant cells, insect cells and animal cells.  
     
     
         10 . An isolated kinase polypeptide comprising an amino acid sequence selected from the group consisting of: 
 (a) amino acids 1-275, 25-275, 140-226, or 149-175 of SEQ ID NO:8;    (b) amino acids 1-302, 8-302, 26-302, 34-295, 60-229, 157-183, or 198-229 of SEQ ID NO:9;    (c) amino acids 1-355, 23-279, 124-221, 134-169, or 183-214 of SEQ ID NO:10;    (d) amino acids 1-631, 57-309, 129-254, 175-200, or 215-247 of SEQ ID NO:11;    (e) amino acids 1-311, 14-271, 41-213, 124-161, or 175-206 of SEQ ID NO:12;    (f) amino acids 1-266, 1-261, 25-258, 101-233, 149-175, or 190-226 of SEQ ID NO:13;    (g) amino acids 1-1360, 25-306, 25-258, 101-233, 149-175, or 190-226 of SEQ ID NO:14;    (h) an amino acid sequence that is at least 80% identical to any of the amino acid sequences of (a)-(g) above.    
     
     
         11 . A method of designing an inhibitor of the polypeptide of  claim 10 , the method comprising the steps of determining the three-dimensional structure of such polypeptide, analyzing the three-dimensional structure for the likely binding sites of substrates, synthesizing a molecule that incorporates a predicted reactive site, and determining the polypeptide-inhibiting activity of the molecule.  
     
     
         12 . A method for identifying compounds that inhibit kinase activity comprising: 
 (a) bringing a test compound into contact with the polypeptide of  claim 10  and a substrate; and    (b) determining whether the test compound inhibits the kinase activity of said polypeptide.    
     
     
         13 . A method for identifying compounds that activate kinase activity comprising: 
 (a) bringing a test compound into contact with the polypeptide of  claim 10  and a substrate; and    (b) determining whether the test compound activates the kinase activity of said polypeptide.    
     
     
         14 . The method of  claim 12  where the test compound is brought into contact with the polypeptide in a cell containing at least one recombinant vector that directs the expression of at least one polynucleotide encoding said polypeptide.  
     
     
         15 . The method of  claim 13  where the test compound is brought into contact with the polypeptide in a cell containing at least one recombinant vector that directs the expression of at least one polynucleotide encoding said polypeptide.  
     
     
         16 . A method for inhibiting the kinase activity of the polypeptide of  claim 10  comprising forming a mixture of the polypeptide of  claim 10 , a substrate, and a compound, wherein the compound blocks the binding of the polypeptide of  claim 10  with the substrate.