Khakimova A K, Skolysheva L K, Shur S A, Vul'fson I L
Biokhimiia. 1995 Feb;60(2):278-88.
Phosphorylase b association with creatine kinase has been studied by frontal elution affinity chromatography, using CNBr-Sepharose 4B immobilized creatine kinase as the affinity matrix. The quantitative parameters of this interaction were estimated from the volumes of phosphorylase b elution at various concentrations of the enzyme. The dissociation constants for phosphorylase b complexes with immobilized creatine kinase and of the phosphorylase b complex with free creatine kinase were found to be equal to 0.49 and 0.191 microM, respectively. In the presence of AMP the interaction between the proteins became weaker. With a rise in AMP concentration from 0.02 to 0.15 mM the value of the dissociation constants increased from 1.59 up to 9.66 microM. One molecule of AMP was shown to bind on the phosphorylase b-immobilized creatine kinase complex.
利用固定在溴化氰活化的琼脂糖4B上的肌酸激酶作为亲和基质，通过前沿洗脱亲和色谱法研究了磷酸化酶b与肌酸激酶的结合。根据不同浓度酶时磷酸化酶b的洗脱体积估算了这种相互作用的定量参数。发现磷酸化酶b与固定化肌酸激酶形成的复合物以及磷酸化酶b与游离肌酸激酶形成的复合物的解离常数分别为0.49和0.191微摩尔。在有AMP存在时，蛋白质之间的相互作用变弱。随着AMP浓度从0.02毫摩尔升至0.15毫摩尔，解离常数的值从1.59微摩尔增加至9.66微摩尔。已表明一分子的AMP结合在磷酸化酶b -固定化肌酸激酶复合物上。
