Purification and characterization of anti-clotting protein component （ACPF-7221） from venom of Agkistrodon acutus

Background Snake venom contains a number of components with different pharmacological and biological activities, especially in cancer therapy, and has increasingly become a research focus. This study was designed to isolate and purify a novel anti-clotting protein component from the venom of Agkistr...

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Bibliographic Details
Published in:Chinese medical journal 2009, Vol.122 (18), p.2169-2173
Main Author: RUI Jing HUAI Jian-guo ZHANG Ye CHENG Dong-yun PAN Xue-bing
Format: Article
Language:English
Subjects:
分离纯化
基金会
尖吻蝮蛇
抗凝血因子
活化部分凝血活酶时间
蛋白成分
预防犯罪
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Summary:Background Snake venom contains a number of components with different pharmacological and biological activities, especially in cancer therapy, and has increasingly become a research focus. This study was designed to isolate and purify a novel anti-clotting protein component from the venom of Agkistrodon acutus, and to explore its physico-chemical properties and biological activity. Methods The venom of Agkistrodon was isolated and purified by ion-exchange chromatography on diethylaminoethyl （DEAE）-Sepharose Fast Flow, molecular sieve filtration through Sephadex G75, SP-Sepharose Fast Flow and molecular sieve filtration through Sephadex G50. We detected the activated partial thromboplastin time （APTT） of the eluant to select the anti-clotting protein component of interest. The molecular weight was determined by sodium dodecyl sulfate-polyacrylamid gel electrphoresis （SDS-PAGE） and liquid chromatography. Its protein content was detected by bicinchoninic acid （BCA）. Results SDS-PAGE vertical gel electrophoresis showed that the anticoagulant factor is a tripolymer composed of three proteins whose molecular weights are 25 KDa, 30 KDa and 50 KDa. The factor contains about 65% percent protein. Conclusions A novel anti-clotting protein component was purified by ion-exchange chromatography and molecular sieve filtration from the venom of Agkistrodon acutus and was found to be composed of three kinds of proteins.
ISSN:0366-6999
2542-5641