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, use...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.展开更多
文摘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.
