日本蝮蛇蛇毒碱性磷脂酶A_2同源物的分离及鉴定  

Purification and characterization of phospholipase A_2 homologue from the manushi (Agkistrodon blomhoffii ussurensis) snake venom

作　　者：杨巍[1] 包永明[1] 段延龙[1] 安利佳[1] 

机构地区：[1]大连理工大学生物工程系,大连116024

出　　处：《动物学报》2003年第5期704-707,共4页ACTA ZOOLOGICA SINICA

摘　　要：We purified and characterizated a phospholipase A 2 homologue from Agkistrodon blomhoffii ussurensis snake venom. We used Hitrap SP cation exchange and Superdex 75 columns chromatography to obtain a basic protein, used SDS PAGE 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 A 2 activity on egg yolk phospholipids, hemolytic activity on washed erythrocytes, and anticoagulant effect on pig platelet rich plasma, as well as the N terminal sequence with protein sequencer. The results showed that it had no phospholipase A 2 activity and hemolytic activity, but had obvious anticoagulant effect on in vitro. The N terminal sequence (21 amino acid residues) compared with other phospholipases A 2 demonstrated that the protein was homogenous with BPLA 2s from Agkistrodon halys Palls.We purified and characterizated a phospholipase A 2 homologue from Agkistrodon blomhoffii ussurensis snake venom. We used Hitrap SP cation exchange and Superdex 75 columns chromatography to obtain a basic protein, used SDS PAGE 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 A 2 activity on egg yolk phospholipids, hemolytic activity on washed erythrocytes, and anticoagulant effect on pig platelet rich plasma, as well as the N terminal sequence with protein sequencer. The results showed that it had no phospholipase A 2 activity and hemolytic activity, but had obvious anticoagulant effect on in vitro. The N terminal sequence (21 amino acid residues) compared with other phospholipases A 2 demonstrated that the protein was homogenous with BPLA 2s from Agkistrodon halys Palls.

关 键 词：日本蝮蛇 碱性磷脂酶A2 蛇毒 抗凝血活性 

分 类 号：Q592[生物学—生物化学]