福建产圆斑蝰蛇(Vipera russelli siamensis)蛇毒磷酯酶A的分离纯化及部分性质研究

THE PURIFICATION AND SOME PROPERTIES OF THE PHOSPHOLIPASE A FROM THE VENOM OF VIPERA RUSSELLI SIAMENSIS(FROM FUJIAN)

采用羧甲基葡聚糖C-50柱层析、葡聚糖G-75凝胶过滤和二乙氨基乙基葡聚糖A-50柱层析三步分离程序,获得了一个聚丙烯酰胺凝胶盘电泳纯的毒性磷酯酶A。用葡聚糖凝胶过滤法测得分子量为18,000,用SDS-平板聚丙烯酰胺凝胶梯度电泳法测得分子量为15,000,用聚丙烯酰胺凝胶等电聚焦法测得等电点为7.6。纯化的毒性磷酯酶A对昆明品系小白鼠的LD;(腹腔注射)是2微克/每克体重。磷酯酶A的比活性为24微克分子/分/毫克。纯化成分的产率是1—1.2%。

A toxie phospholipase A was purified from the venom of vipera russelli siamensis by ion-exchange chromatography on CM-Sephadex C-50,gel filtration on Sephadex G-75 and ion-exchange chromatography on DEAE-Sephadex A-50.A homogeneous phospholipase A was exhibited by polyacrylamide gel electrophoretic criterion.Its molecular weight,calculated from measurements in Sephadex column chromatography and polyacrylamide gel elcctrophoresis,is 18,000 and 15,000 respectively.The isoeiectric point is 7.6 by isoelcclric focusing.The toxic phospholipase A has an acute LD100 values(intraperitoneal injection)of 20u,g.per 202gm mouse.The phospholipase A activity was measured by titration of the fatty acids released from the lecithin using an automatic pH stat apparatus.The specific activity is 24 umol./min/mg.The yield of the toxic phospholipase A is between 1 and 1.2%oi dried venom.