Guliaev N N, Tunitskaia V L, Baranova L A, Nesterova M V, Murtuzaev I M
Biokhimiia. 1976;41(7):1241-9.
A number of unknown ATP analogues is isolated when studying the structure of the active site of catalytic histonekinase subunit. Adenosine-5'-chloromethanepyrophosphonate adenosine-5'-(beta-bromoethanepyrophosphonate) and adenosine-5'-(p-fluorosulphonylphenylphosphate) were isolated under the reaction of chloromethanephosphonic acid, beta-bromoethanephosphonic acid and n-phenolsulphofluoride respectively with AMP imidazolide. Adenosine-5'-(beta-chloroethylphosphate) was obtained from AMP morpholide and ethylenechorohydrine. Adenosine-5'-chloracetylaminomethanephosphonate and adenosine-5'-(p-fluorosulphonylbenzoylaminomethanephosphonate) were obtained in the reaction of chloroacetyc anhydride and n-fluorosulphonylbenzoylchloride. Adenosine-5'-(p-aminophenylphosphate) is synthesized under the reduction of AMP mononitrophenyl ester. The treatment of the former with chloroacetyc anhydride produced adenosine-5'-(p-chloroacetylaminophenylphosphate. Interaction of ATP analogues obtained and also of early synthesized adenosine-5'-chloromethanephosphonate and adenosine-5'-(beta-bromoethanephosphonate) with homogenous catalytic histonekinase subunit is studied. The decrease in the reaction rate of Hi histone phosphorylation is found to take place. pH optimum of the enzyme inactivation with adenosine-5'-chloromethanepyrophosphonate and adenosine-5'-(beta-chloroethylphosphate) and the protective effect of the substrate (ATP) indicate covalent blocking imidazole ring in the active site. The date obtained suggest that the functional group of the active site of catalytic histonekinase subunit is histidine imidazole ring located close to terminal ATP phosphate.
在研究催化组蛋白激酶亚基活性位点的结构时，分离出了多种未知的ATP类似物。分别在氯甲膦酸、β-溴乙膦酸和对氟磺酰基苯氟化物与AMP咪唑化物的反应中，分离出了腺苷-5'-氯甲基焦磷酸酯、腺苷-5'-（β-溴乙基焦磷酸酯）和腺苷-5'-（对氟磺酰基苯基磷酸酯）。腺苷-5'-（β-氯乙基磷酸酯）是由AMP吗啉化物和氯乙醇得到的。在氯乙酰酐和对氟磺酰基苯甲酰氯的反应中，得到了腺苷-5'-氯乙酰氨基甲基膦酸酯和腺苷-5'-（对氟磺酰基苯甲酰氨基甲基膦酸酯）。在AMP单硝基苯酯还原反应中合成了腺苷-5'-（对氨基苯基磷酸酯）。用氯乙酰酐处理前者得到腺苷-5'-（对氯乙酰氨基苯基磷酸酯）。研究了所得ATP类似物以及早期合成的腺苷-5'-氯甲基膦酸酯和腺苷-5'-（β-溴乙基膦酸酯）与均一的催化组蛋白激酶亚基的相互作用。发现组蛋白H1磷酸化反应速率降低。腺苷-5'-氯甲基焦磷酸酯和腺苷-5'-（β-氯乙基磷酸酯）使酶失活的最适pH值以及底物（ATP）的保护作用表明活性位点中的咪唑环被共价阻断。所得数据表明，催化组蛋白激酶亚基活性位点的官能团是靠近末端ATP磷酸基团的组氨酸咪唑环。
