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 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.
Department of Surgery, Yijishan Hospital, Wannan Medical College, Wuhu, Anhui 241001, China. ruijing1956@sina.com
Jing
J
Rui
Jian-guo
JG
Huai
Ye
Y
Zhang
Dong-yun
DY
Cheng
Xue-bing
XB
Pan
D017836:Agkistrodon / Q000378:metabolism*; D000818:Animals; D000925:Anticoagulants / Q000737:chemistry / Q000302:isolation & purification*; D002851:Chromatography, High Pressure Liquid; D002852:Chromatography, Ion Exchange; D003435:Crotalid Venoms / Q000737:chemistry*; D004591:Electrophoresis, Polyacrylamide Gel; D011506:Proteins / Q000737:chemistry / Q000302:isolation & purification*
D000925:Anticoagulants; D003435:Crotalid Venoms; D011506:Proteins