Fister P, Eigenbrodt E, Presek P, Reinacher M, Schoner W
Biochem Biophys Res Commun. 1983 Sep 15;115(2):409-14. doi: 10.1016/s0006-291x(83)80159-4.
Isolated hepatocytes from 24 h starved chicks were depleted of phosphate and incubated in the presence of 32P-orthophosphate. Pyruvate kinase type M2 from crude cell extracts was partially purified by chromatography on DEAE-Sephacel and hydroxylapatite. SDS slab gel electrophoresis of the fractions containing the enzyme and immunoprecipitation with antisera showed the phosphorylation of pyruvate kinase type M2. Phosphoamino acid analysis identified serine as phosphate acceptor.
从饥饿24小时的雏鸡中分离出的肝细胞被耗尽磷酸盐,并在32P-正磷酸盐存在下孵育。粗细胞提取物中的丙酮酸激酶M2型通过在DEAE-琼脂糖凝胶和羟基磷灰石上进行色谱法进行部分纯化。对含有该酶的级分进行SDS平板凝胶电泳并用抗血清进行免疫沉淀,结果显示丙酮酸激酶M2型发生了磷酸化。磷酸氨基酸分析确定丝氨酸为磷酸受体。