Grivennikov I A, Bulargina T V, Khropov Iu V, Guliaev N N, Severin S E
Biokhimiia. 1979 May;44(5):771-80.
A method for the preparation of a homogenous catalytic subunit of adenosine 3':5'-monophosphate-dependent protein kinase from pigeon breast muscle was developed. The molecular weight of the enzyme as determined by electrophoresis in the presence of sodium dodecyl sulfate was found to be 42000. The pH optimum of the catalytic subunit was around 8.0. The active site of the catalytic subunit was studied using some derivatives of ATP, containing different reactive groups in the triphosphate chain of the molecule. It may be assumed that the pH optimum of the enzyme inactivation by adenosine 5'-chloromethylpyrophosphonate and the protective effect of ATP suggest covalent binding of the imidazole ring in the enzyme active site. The kinetic mechanism of the protein kinase reaction was studied using the initial rate experiments and reaction product inhibition. The results obtained were consistent with a random Bi-Bi kinetic mechanism.
已开发出一种从鸽胸肌中制备腺苷3':5'-单磷酸依赖性蛋白激酶均一催化亚基的方法。在十二烷基硫酸钠存在下通过电泳测定，该酶的分子量为42000。催化亚基的最适pH约为8.0。使用ATP的一些衍生物研究了催化亚基的活性位点，这些衍生物在分子的三磷酸链中含有不同的反应基团。可以假定，5'-氯甲基焦磷酸腺苷使酶失活的最适pH以及ATP的保护作用表明酶活性位点中的咪唑环发生了共价结合。使用初速率实验和反应产物抑制研究了蛋白激酶反应的动力学机制。所得结果与随机的双底物双产物动力学机制一致。
