浙江蝮蛇毒碱性磷酯酶A的分离纯化及其性质的研究  

ISOLATION,PURIFICATION AND PROPERTIES OF THE BASIC PHOSPHOLIPASE A FROM THE SNAKE VENOM OF AGKISTRODON HALYS PALLAS IN ZHEJIANG

作　　者：武祥福 陈远聪 Wu Xiang-fu;Chen Yuan-chung(Shanghai Institute of Biochemistry,Academia Sinica)

机构地区：[1]中国科学院上海生物化学研究所

出　　处：《Zoological Research》1981年第S01期13-21,共9页动物学研究（英文）

摘　　要：采用DEAE-纤维素柱层析将浙江蝮蛇毒分离后,测得二个磷酯酶A活性峰。其中之一是在用平衡缓冲液最先洗脱下来的,为碱性磷酯酶A。另一是在含0.1MNaCl的缓冲液洗脱下来的,包含有一个酸性磷酯酶A和一个我们在前文已报道的突触前神经毒素。本文报道的是碱性磷酯酶A的纯化及其性质的研究。碱性磷酯酶A进一步经磷酸-纤维素,CM-Sephadex C-50和Sephadex G-75柱层析纯化,得到一个在聚丙烯酰胺凝胶电泳上为一条区带的单一蛋白。其分子量为13,800左右,由121个氨基酸残基组成,富含半胱氨酸(14个),酪氨酸,甘氨酸和脯氨酸,等电点为9.3。如同其他蛇毒磷酯酶A的性质相似,此碱性磷酯酶A显示很好的热稳定性。胰蛋白酶不能使它的酶活性完全丧失。Two peaks with phospliolipasc A activity were found by DEAE-ccllulose chromatography from the snake venom ot Agkislrodon halys Pallas in Zhe-jiang.One of them was first eluted by the equilibrium buffer,which was the basic phospliolipasc A.Another one eluiecl by the buffer containing 0.1M NaCl is composed of an acidic phospholipase A and the prcsynaptic ncuroloxin,which had been reported in the preceding paper.In this paper,we report the purification and properties of the basic phospholipase A.The enzyme was further purified by P-ccllulose,CM-Sephadex C-50 and Sephadcx G-75 chromatographics.This purified protein appeared to be a single band on polyacrylamide gel clectrophoresis.Its molecular weight is about 13,800,composed of 121 amino acid residues,rich in l/2cystinc(14),Tyrosine,glycine and proline,its isoelectric point is 9.3.Like the other phospholipase A found in snake venoms,the basic phospholipase A exhibits extreme heat-stability.It remains part of its activity by treating with trypsin.

关 键 词：蝮蛇毒 突触前神经毒素 磷酯酶 柱层析 聚丙烯酰胺凝胶电泳 胰蛋白酶 氨基酸残基 半胱氨酸 

分 类 号：R28[医药卫生—中药学]