Investigation of the adenosine 3', 5'-cyclic phosphate binding site of pig brain histone kinase with the aid of some analogues of adenosine 3', 5'-cyclic phosphate.
2'-O-Chloroacetyl cyclic AMP, 2'-O-acrylyl cyclic AMP and N-6, 2'-O-diacrylyl cyclic AMP were synthesized by the reaction of cyclic AMP with chloroacetic and acrylic anhydrides, respectively.  Selective O-deacylation of N-6, 2'-O-diacrylyl cyclic AMP yielded N-6 -monoacrylyl cyclic AMP.  In the reaction of gamma-mercaptobutyric acid with 8-bromo cyclic AMP, 8-(gamma-carboxypropylthio) cyclic AMP was obtained.  The compounds synthesized and other cyclic AMP analogues (8-bromo cyclic AMP and adenosine 3', 5'-cyclic sulphate) were tested for ability to interact with the highly purified pig brain histone kinase.  All compounds under study were found to be activators of the enzyme.  The highest activating potency was manifested by 8-bromo cyclic AMP and 8-(gamma-carboxypropylthio) cyclic AMP; adenosine 3', 5'-cyclic sulphate was the least potent in this respect.  All compounds were shown to inhibit binding of cyclic [-3-H]AMP to histone kinase.  The inhibition was competitive with respect to cyclic AMP in all cases.  All compounds, except for 2'-O-chloroacetyl cyclic AMP may indicate the formation of a covalent bond between this analogue and the enzyme.  These findings suggest that an active site of the regulatory subunit of the histone kinase contains at least three specific areas responsible for cyclic AMP binding.
D000179:Acrylates; D000818:Animals; D001665:Binding Sites; D001921:Brain / Q000201:enzymology*; D000242:Cyclic AMP / Q000031:analogs & derivatives / Q000378:metabolism*; D007700:Kinetics; D006656:Protamine Kinase / Q000378:metabolism*; D011485:Protein Binding; D011494:Protein Kinases / Q000378:metabolism*; D013329:Structure-Activity Relationship; D013464:Sulfuric Acids; D013552:Swine
D000179:Acrylates; D013464:Sulfuric Acids; D000242:Cyclic AMP; D011494:Protein Kinases; D006656:Protamine Kinase