Kits and peptides for transferases

Abstract

Kits and peptide substrates for detecting transferase activity of a sample are provided. The kits include a substrate including a reporter compound, at least one of a phosphate group donor and a phosphate group acceptor, a buffer that supports enzymatic activity of the transferase, and a peptidase that cleaves a non-phosphorylated peptide substrate at a first rate and a phosphorylated peptide substrate at a second rate. The peptide substrates include a reporter compound and a first transferase substrate linked to the reporter compound on a first side of the reporter compound.

1 . A kit for determining transferase activity, the kit comprising:
 (A) a substrate including a reporter compound;   (B) at least one of a phosphate group donor and a phosphate group acceptor;   (C) a buffer that supports enzymatic activity of the transferase; and   (D) a peptidase that cleaves a non-phosphorylated peptide substrate at a first rate and a phosphorylated peptide substrate at a second rate.   
     
     
         2 . A kit of  claim 1 , wherein the substrate comprises a substrate for a kinase. 
     
     
         3 . A kit of  claim 1 , wherein the kit further comprises a transferase. 
     
     
         4 . A kit of  claim 1 , wherein the substrate comprises a substrate for a phosphate. 
     
     
         5 . A kit of  claim 1 , wherein the peptidase comprises an aminopeptidase. 
     
     
         6 . A kit of  claim 1 , wherein the reporter compound comprises a fluorogenic compound. 
     
     
         7 . A kit of  claim 1 , where in the reporter compound comprises a luminogenic compound. 
     
     
         8 . A peptide substrate comprising:
 (A) a reporter compound; and   (B) a first transferase substrate linked to the reporter compound on a first side of the reporter compound.   
     
     
         9 . A peptide substrate of  claim 8 , wherein the first transferase substrate is a phosphotyrosine. 
     
     
         10 . A peptide substrate of  claim 8 , further comprising a second transferase substrate linked to the reporter compound on a second side of the reporter compound, wherein reporter compound is Rhodamine 110, and wherein the first and second transferase substrates are phosphotyrosine. 
     
     
         11 . A peptide substrate of  claim 8 , further comprising a second transferase substrate linked to the reporter compound on a second side of the reporter compound, wherein reporter compound is Rhodamine 110, and wherein the first and second transferase substrate are AAY(PO 3 )AXAA (SEQ. ID. NO:34). 
     
     
         12 . A peptide substrate of  claim 8 , further comprising a second transferase substrate linked to the reporter compound on a second side of the reporter compound, wherein reporter compound is Rhodamine 110, and wherein the first and second transferase substrates are LRRASLG (SEQ. ID. NO:1). 
     
     
         13 . A peptide substrate of  claim 8 , further comprising a second transferase substrate linked to the reporter compound on a second side of the reporter compound, wherein reporter compound is Rhodamine 110, and wherein the first and second transferase substrates are KKALRRASLKG (SEQ. ID. NO:2). 
     
     
         14 . A peptide substrate of  claim 8 , further comprising a second transferase substrate linked to the reporter compound on a second side of the reporter compound, wherein reporter compound is Rhodamine 110, and wherein the first and second transferase substrates are KKALRKASVRG (SEQ. ID. NO:3). 
     
     
         15 . A peptide substrate of  claim 8 , further comprising a second transferase substrate linked to the reporter compound on a second side of the reporter compound, wherein reporter compound is Rhodamine 110, and wherein the first and second transferase substrates are YIYGAFKRRG (SEQ. ID. NO:4). 
     
     
         16 . A peptide substrate of  claim 8 , further comprising a second transferase substrate linked to the reporter compound on a second side of the reporter compound, wherein reporter compound is Rhodamine 110, and wherein the first and second transferase substrates are RRAT(PO 3 )VA (SEQ. ID. NO:5). 
     
     
         17 . A peptide substrate of  claim 8 , further comprising a second transferase substrate linked to the reporter compound on a second side of the reporter compound, wherein reporter compound is Rhodamine 110, and wherein the first and second transferase substrates are YIY(PO 3 )GAFKRRG (SEQ. ID. NO:6). 
     
     
         18 . A peptide substrate of  claim 8 , wherein the reporter compound is 7-amino-4-methylcoumarin, and wherein the first transferase substrate is LRRASLG (SEQ. ID. NO:1).