日本蝮蛇蛇碱性磷脂酶A同源物的分离及鉴定.pdfVIP

动物学报 　49 (5) :704～707 , 2003 A cta Zoologica S inica 　　　　　 　 日本蝮蛇蛇毒碱性磷脂酶A2 同源物的分离及鉴定 杨　巍　包永明 　段延龙　安利佳 (大连理工大学生物工程系 , 大连　116024) Purif ication and characterization of phospholipase A2 homologue from the manushi ( Agkistrodon blomhof f ii ussurensis) snake venom YAN G Wei 　BAO YongMing 　DUAN YanLong 　An LiJia ( ) Depart ment of Biochemical Engi neeri ng , Dalian U niversity of Technology , Dalian 　116024 , Chi na Abstract 　We purified and characterizated a phospholipase A2 homologue from A gkist rodon blom hof f ii ussurensis snake venom. We used Hitrap SP cation exchange and Superdex 75 columns chromatography to obtain a basic protein , used SDSPA GE to analyse molecular mass , and IEF ( Isoelectric focusing electrophoresis) IEF to identify isoelectric point. The molecular mass was 16 kDa , and the isoelectric point was 8 56. We detected its phospholipase A2 activity on egg yolk phospholipids , hemolytic activity on washed erythrocytes , and anticoagulant effect on pig plateletrich plasma , as well as the Nterminal sequence with protein sequencer. The results showed that it had no phospholipase A2 activity and hemolytic activity , but had obvious anticoagulant effect on in vit ro. The Nterminal sequence (21 amino acid residues) compared with other phospholipases A demonstrated that the protein was homogenous with BPLA s from A gkist rodon halys Palls