纯化的磷脂酰肌醇特异性磷脂酶C（金黄色葡萄球菌）对红细胞膜的修饰
Modification of erythrocyte membranes by a purified phosphatidylinositol-specific phospholipase C (Staphylococcus aureus).
作者信息
Low M G, Finean J B
出版信息
Biochem J. 1977 Feb 15;162(2):235-40. doi: 10.1042/bj1620235.
A phosphatidylinositol-specific phospholipase C from Staphylococcus aureus was purified by a three-step procedure. The specific activity of the purified enzyme was approx. 6000 times that of the culture supernatant, with an overall recovery of approx. 10%. Estimation of the molecular weight by sodium dodecyl sulphate/polyacrylamide-gel electrophoresis and by gel filtration gave values of 33000 and 20000 respectively. A thiol group appears to be necessary for the activity of the enzyme. The purified enzyme had no detectable delta-haemolytic activity and was unable to hydrolyse S. aureus phospholipids. Phosphatidyl-inositol in erythrocyte 'ghosts' was readily hydrolysed by the purified phospholipase C. However, in contrast with our previous preliminary observations, phosphatidylinositol in intact erythrocytes was not significantly hydrolysed. These results suggest that at least 75-80% of the phosphatidylinositol is located at the inner leaflet of the membrane.
通过三步程序纯化了来自金黄色葡萄球菌的磷脂酰肌醇特异性磷脂酶C。纯化酶的比活性约为培养上清液的6000倍，总回收率约为10%。通过十二烷基硫酸钠/聚丙烯酰胺凝胶电泳和凝胶过滤法估算分子量，分别得到33000和20000的值。硫醇基团似乎是该酶活性所必需的。纯化的酶没有可检测到的δ溶血活性，并且无法水解金黄色葡萄球菌的磷脂。纯化的磷脂酶C能轻易水解红细胞“血影”中的磷脂酰肌醇。然而，与我们之前的初步观察结果相反，完整红细胞中的磷脂酰肌醇没有被显著水解。这些结果表明，至少75 - 80%的磷脂酰肌醇位于膜的内小叶。
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