Lapetina E G, Seguin E B, Agranoff B W
Biochim Biophys Acta. 1975 Jul 22;398(1):118-24. doi: 10.1016/0005-2760(75)90175-7.
A method is described for the preparation of radioactive inositol lipids for studies of their enzymic degradation. Kidney cytosol fractions have been used to produce diesteratic cleavage. High voltage electrophoresis at pH 4.3 is used to separate D-myoinositol 1 : 2-cyclic phosphate and D-myoinositol 1-phosphate from hydrolysis of phosphatidylinositol. Radioactivity co-migrating with myoinositol diphosphate and triphosphate is separated by electrophoresis at pH 1.5 following enzymatic hydrolysis of phosphatidylinositol phosphate and phosphatidylinositol diphosphate. Relative activities for hydrolysis of the various inositides suggest the presence of more than one phosphodiesterase.
本文描述了一种制备放射性肌醇脂质以研究其酶促降解的方法。已使用肾细胞溶质部分进行二酯键裂解。在pH 4.3条件下进行高压电泳，以从磷脂酰肌醇的水解产物中分离出D-肌醇1:2-环磷酸酯和D-肌醇1-磷酸酯。在对磷脂酰肌醇磷酸酯和磷脂酰肌醇二磷酸酯进行酶促水解后，通过在pH 1.5条件下进行电泳，将与肌醇二磷酸酯和三磷酸酯共迁移的放射性物质分离出来。各种肌醇磷脂水解的相对活性表明存在不止一种磷酸二酯酶。
