Human receptor tyrosinge kinase

Abstract

An isolated and purified human receptor tyrosine kinase is described. A cDNA sequence which encodes the native polypeptide is disclosed as well as the structural coding region and the amino acid residue sequence of the tyrosine kinase. Methods are provided which employ novel sequences to identify compounds that modulate the biological and/or pharmacological activity of the tyrosine kinase and hence regulate cellular and tissue physiology. Biologically-effective antisense molecules, as well as dominant negative mutant versions of the kinase, are described which are suitable for therapeutic use. The invention is also related to the diagnosis, study, prevention, and treatment of pathophysiological disorders related to or mediated by the biological molecule.

1 . A purified polynucleotide comprising a nucleic acid sequence which encodes a polypeptide having at least about 80% homology to a member selected from the group consisting essentially of: (SEQ ID NO:3, SEQ ID NO:3 positions 1-25, SEQ ID NO:3 positions 1-122, SEQ ID NO:3 positions 26-422, and SEQ ID NO:3 positions 123-422).  
     
     
         2 . A polynucleotide according to  claim 1  comprising a nucleic acid sequence which encodes a polypeptide comprising the sequence as depicted in SEQ ID NO:3.  
     
     
         3 . A polynucleotide according to  claim 1  comprising a nucleic acid sequence which encodes a polypeptide comprising the sequence as depicted in SEQ ID NO:3 wherein position 147 (Lysine) of SEQ ID NO:3 is substituted or deleted.  
     
     
         4 . A polynucleotide according to  claim 3  comprising a nucleic acid sequence which encodes a polypeptide comprising the sequence as depicted in SEQ ID NO:3 wherein position 147 of SEQ ID NO:3 is Arginine.  
     
     
         5 . A polynucleotide according to  claim 1  comprising a nucleic acid sequence which encodes a polypeptide comprising a sequence selected from the group consisting essentially of: (SEQ ID NO:3 positions 1-25, SEQ ID NO:3 positions 1-122, SEQ ID NO:3 positions 26-422, and SEQ ID NO:3 positions 123-422).  
     
     
         6 . A polynucleotide of  claim 2  wherein the polynucleotide sequence comprises the sequence as depicted in SEQ ID NO:2.  
     
     
         7 . An antisense molecule comprising an oligomer in the range from about 12 to about 25 nucleotides in length which: (a) is complementary to a region within positions 67-148 of SEQ ID NO:1; or, (b) comprises a sequence which is complementary to a sequence selected from the group consisting of: (SEQ ID NO:1 positions 67-79; 70-82; 73-85; 75-92; 76-88; 79-91; 80-92; 81-93; 82-94; 83-95; 84-96; 85-97; 86-98; 87-99; 88-100; 89-101; 90-102; 91-103; 92-104; 93-105; 94-106; 95-107; 96-108; 97-109; 98-110; 99-111; 100-112; 101-113; 102-114; 103-115; 104-116; 105-117; 106-118; 107-119; 108-120; 109-121; 110-122; 111-123; 112-124; 113-125; 114-126; 115-127; 116-128; 117-129; 118-130; 119-131; 120-132; 121-133; 122-134; 123-135; 124-136; 125-137; 126-138; 127-139; 128-140; and 136-148).  
     
     
         8 . An antisense molecule comprising an oligomer in the range from about 12 to about 25 nucleotides in length which: (a) is complementary to a region within positions 1333-1414 of SEQ ID NO:1; or (b) comprises a sequence which is complementary to a sequence selected from the group consisting of: (SEQ ID NO:1 positions 1333-1345; 1336-1348; 1339-1351; 1342-1354; 1345-1357; 1346-1358; 1347-1359; 1348-1360; 1348-1365; 1349-1361; 1350-1362; 1351-1363; 1352-1364; 1353-1365; 1354-1366; 1355-1367; 1356-1368; 1357-1369; 1358-1370; 1359-1371; 1360-1372; 1361-1373; 1362-1374; 1363-1375; 1364-1376; 1365-1377; 1366-1378; 1367-1379; 1368-1380; 1369-1381; 1370-1382; 1371-1383; 1372-1384; 1373-1385; 1374-1386; 1375-1387; 1376-1388; 1377-1389; 1378-1390; 1379-1391; 1380-1392; 1381-1393; 1382-1394; 1383-1395; 1384-1396; 1385-1397; 1386-1398; 1387-1399; 1388-1400; 1389-1401; 1390-1402; 1391-1403; 1392-1404; 1393-1405; 1394-1406; and 1412-1414).  
     
     
         9 . An expression vector comprising the polynucleotide of  claim 1 .  
     
     
         10 . A host cell transformed with the expression vector of  claim 9 .  
     
     
         11 . A purified polypeptide comprising the amino acid sequence as depicted in SEQ ID NO:3 or a variant of SEQ ID NO:3 having at least about 80% homology to a member selected from the group consisting essentially of: (SEQ ID NO:3, SEQ ID NO:3 positions 1-25, SEQ ID NO:3 positions 1-122, SEQ ID NO:3 positions 26-422, and SEQ ID NO:3 positions 123-422).  
     
     
         12 . An antibody specific for the polypeptide SEQ ID NO:3.  
     
     
         13 . A method for producing a polypeptide according to  claim 11 , said method comprising the steps of: 
 a) culturing a host cell according to  claim 10  under conditions suitable for the expression of said polypeptide, and    b) recovering said polypeptide from the host cell culture.    
     
     
         14 . A method of identifying compounds that modulate a biological and/or pharmacological activity of a tyrosine kinase, comprising: 
 a) combining a candidate compound modulator with a polypeptide according to  claim 11 , and    b) measuring an effect of the candidate compound modulator on the biological and/or pharmacological activity of the polypeptide.    
     
     
         15 . A method of identifying compounds that modulate a biological and/or pharmacological activity of a tyrosine kinase according to  claim 14 , comprising: 
 a) combining a candidate compound modulator with a host-cell which expresses said polypeptide, and    b) measuring an effect of the candidate compound modulator on the biological and/or pharmacological activity of the polypeptide.    
     
     
         16 . A compound that modulates the activity of a tyrosine kinase identified by the method of  claim 14 .  
     
     
         17 . A method of treatment of a patient in need of such treatment for a condition which is mediated by a tyrosine kinase, comprising administering an effective amount of a compound according to  claim 16 .  
     
     
         18 . A method of modulating a biological and/or pharmacological activity of a tyrosine kinase in a cell comprising administering an effective amount of a polynucleotide according to  claim 1  to said cell.  
     
     
         19 . A method for modulating the expression of a tyrosine kinase in a cell comprising administering an effective amount of an antisense molecule according to  claim 7  or  claim 8  to said cell.  
     
     
         20 . A diagnostic composition, for the identification of a polypeptide comprising the amino acid sequence as depicted in SEQ ID NO:3, comprising the antibody of  claim 12 .  
     
     
         21 . A diagnostic composition for the identification of a polynucleotide sequence comprising PCR primers derived from SEQ ID NO:1 or allele sequences representative of 11, 12, 13 or 15 GT repeats between positions 2125 and 2152 of SEQ ID NO:1.