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.
Rui
Jing
J
Department of Surgery, Yijishan Hospital, Wannan Medical College, Wuhu, Anhui 241001, China. ruijing1956@sina.com
Huai
Jian-guo
JG
Zhang
Ye
Y
Cheng
Dong-yun
DY
Pan
Xue-bing
XB
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