The purification and identification of phospholipid-binding anticoagulation protein from agkistrodon halys brevicaudus venom
- VernacularTitle:短尾蝮蛇毒中磷脂结合抗凝蛋白的分离纯化及鉴定
- Author:
Yuanhong LI
;
Zhaoyan LI
;
Faquan LIN
;
Binglun HUANG
- Publication Type:Journal Article
- Keywords:
purify;
anticoagulation;
phospholipid;
venom
- From:
Chinese Pharmacological Bulletin
2003;0(07):-
- CountryChina
- Language:Chinese
-
Abstract:
Aim To purify Phospholipid-binding anticoagulation protein(PBAP) from Agkistrodon halys Brevicaudus Venom and study the biochemical characterization.Methods The Phospholipid-binding anticoagulation protein was purified from Agkistrodon halys Brevicaudus Venom by Cation ion exchange chromatography on CM Sephadex C-25 and negative ion exchange chromatography on DEAE Sepharose CL-6B,gel filtration on Sephacryl S-200 and Sephadex G-75 chromatography.Its anticoagulant activities in vitro were assayed by activated partial thromboplastin time(APTT);its molecular weight was calculated by SDS-polyacrylamide gel electrophoresis(SDS-PAGE) and its isoelectric point was estimated by the isoelectric focusing electrophoresis.Binding experiments of anticoagulation protein to phospholipids vesicles were performed with thinlayer chromatography.Results A kind of protein was purified from Agkistrodon halys Brevicaudus Venom which was able to prolong APTT.The SDSPAGE showed that it was dimer and its molecular weight was 24.0?10~3 under non-reducing condition and 14.6?10~3 under reducing condition.The isoelectric point was pH 5.2 by the isoelectric focusing electrophoresis.Having arginine ester-hydrolyzing enzyme and binding Phospholipid activify,its effect on APTT was activity stronger with concentration increasing.Conclusion It is a successful method of the purification of Phospholipid-binding anticoagulation protein from the Agkistrodon halys Brevicaudus Venom.
