Materials and Methods Related to Sodium/Potassium Adenosine Triphosphatase and SRC

Abstract

Described herein are methods to bind a compound to the SH2 domain of Src in a Src-expressing cell which includes contacting a compound comprising an amino acid compound to a Src-expressing cell; and binding a compound to the SH2 domain of Src.

We claim:  
     
         1 . A method to bind a compound to the SH2 domain of Src in a Src-expressing cell comprising:
 a) contacting a compound comprising an amino acid compound consisting of the sequence STNCV EGTAR GIVVY TGD [SEQ ID NO: 1] to a Src-expressing cell; and   b) binding a compound to the SH2 domain of Src.   
     
     
         2 . The method of  claim 1 , wherein the Src-expressing cell is a mammalian cell. 
     
     
         3 . The method of  claim 1 , wherein the mammalian cell is a monocyte. 
     
     
         4 . The method of  claim 1 , wherein the at least one mammalian cell is a cell selected from the group consisting of: heart cell; liver cell; vascular cell; breast cell; prostate cell; kidney cell; muscle cell; blood cell; and brain cell. 
     
     
         5 . The method of  claim 1 , wherein the at least one mammalian cell is cultured in vitro. 
     
     
         6 . The method of  claim 1 , wherein the at least one mammalian cell is in an animal model. 
     
     
         7 . The method of  claim 1 , wherein the at least one mammalian cell is human. 
     
     
         8 . A method of treating a Src-associated disease in a mammal in need of such treatment comprising:
 a) administering to a mammal a therapeutic composition comprising an amino acid compound consisting of the sequence STNCV EGTAR GIVVY TGD [SEQ ID NO: 1]; and   b) treating a Src-associated disease in the mammal.   
     
     
         9 . The method of  claim 8 , wherein the mammal is a human. 
     
     
         10 . The method of  claim 8 , wherein the Src-associated disease is selected from the group consisting of: cancer; vascular disease; cardiovascular disease; heart disease; prostate cancer; breast cancer; neuroblastoma; cardiac hypertrophy; tissue fibrosis; congestive heart failure; and ischemia/reperfusion injury. 
     
     
         11 . A method of treating cancer in a mammal in need of such treatment comprising:
 a) administering to a mammal a composition comprising an amino acid compound consisting of the sequence STNCV EGTAR GIVVY TGD [SEQ ID NO: 1]; and   b) treating cancer in the mammal.   
     
     
         12 . The method of  claim 11 , further comprising administering to the mammal at least one additional therapeutic useful to treat cancer. 
     
     
         13 . The method of  claim 11 , wherein the composition inhibits growth of the cancer. 
     
     
         14 . The method of  claim 11 , wherein the composition reduces an increased basal Src activity in the cancer. 
     
     
         15 . The method of  claim 11 , wherein the composition reduces migration of the cancer. 
     
     
         16 . The method of  claim 11 , wherein the composition causes cell death of the cancer. 
     
     
         17 . The method of  claim 11 , wherein the cancer is selected from the group consisting of: breast cancer; prostate cancer; and neuroblastoma. 
     
     
         18 . A method of treating a cardiovascular disease in a mammal in need of such treatment comprising:
 a) administering to a mammal a composition comprising an amino acid compound consisting of the sequence STNCV EGTAR GIVVY TGD [SEQ ID NO: 1]; and   b) treating a cardiovascular disease in the mammal.   
     
     
         19 . The method of  claim 18 , further comprising administering to the mammal at least one additional therapeutic useful to treat cardiovascular disease. 
     
     
         20 . The method of  claim 18 , wherein the cardiovascular diseases is selected from the group consisting of: cardiac hypertrophy; congestive heart failure; heart disease; and ischemia/reperfusion; vascular disease; and atherosclerosis. 
     
     
         21 . A method of treating tissue fibrosis in a mammal in need of such treatment comprising:
 a) administering to a mammal a composition comprising an amino acid compound consisting of the sequence STNCV EGTAR GIVVY TGD [SEQ ID NO: 1]; and   b) treating a cardiovascular disease in the mammal.   
     
     
         22 . The method of  claim 21 , further comprising administering to the mammal at least one additional therapeutic useful to treat tissue fibrosis. 
     
     
         23 . A method to screen for test compositions capable of inhibiting binding between CD2 domain of Na/K ATPase and SH2 domain of Src, comprising the steps of:
 a) incubating test compositions with purified Src for at least 5 and up to 45 minutes in a vessel;   b) introducing Na/KATPase to the vessel for at least 5 minutes and up to 45 minutes;   c) introducing ouabain to the vessel for at least 1 minute and up to 45 minutes;   d) introducing Mg2+/ATP to the vessel for at least 0.5 minute and up to 45 minutes;   e) conducting western blot analysis to determine if Src activation was induced by ouabain.   
     
     
         24 . The method of  claim 23 , wherein steps a) and b) are each about 12 minutes to about 17 minutes, step c) is about 2 minute to about 7 minutes, and step d) is at least one minute. 
     
     
         25 . A method to screen for test compositions capable of inhibiting binding between CD2 domain of Na/K ATPase and SH2 domain of Src, comprising the steps of:
 a) contacting a test composition with a FRET pair of CD2 domain and SH2 domain; and   b) identifying those compositions which abolish the FRET energy as capable of inhibiting the CD2/SH2 interaction.   
     
     
         26 . The method of  claim 25 , wherein the CD2 is labeled with Cy3 and the SH2 is labeled with Cy5.