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

作     者：RUI Jing HUAI Jian-guo ZHANG Ye CHENG Dong-yun PAN Xue-bing 

作者机构：Department of Surgery Yijishan Hospital Wannan Medical College Wuhu Anhui 241001 China Department of Oncology Yijishan Hospital Wannan Medical College Wuhu Anhui 241001 China 

出 版 物：《Chinese Medical Journal》 (中华医学杂志（英文版）)

年 卷 期：2009年第122卷第18期

页      面：2169-2173页

核心收录：

学科分类：1002[医学-临床医学] 100210[医学-外科学(含：普外、骨外、泌尿外、胸心外、神外、整形、烧伤、野战外)] 10[医学] 

基　　金：This study was supported by a grant from the Scientific Research Foundation of the Anhui Committee of Education (No. KJ2007A039) 

主　　题：Agkistrodon acutus anticoagulation purification high performance liquid chromatography 

摘      要：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.