Purification of Blood Coagulation Factor V Activator from the Venom of Iranian Vipera lebetina

Document Type : Original Article

Authors

1 Department of Biochemistry, Faculty of Medicine, Ahvaz Jundishapur University of Medical Sciences, Ahvaz, Iran.

2 Department of Venomous Animals and Antivenom Production, Razi Vaccine and Serum Research Institute, Karaj, Iran.

Abstract

Background and Objective: The  venom  of  many Viperidae  snake appear to contain proteins  that  affect  blood  coagulation. The aim of this study was to identify and isolate a blood coagulation factor V activator present in the venom from Vipera lebetina.
Material  and  Methods:  Factor V activator was purified  from 200 mg of crude venom by three steps  of chromatorgraphy  which included:  gel filtration on sephadex G100,  Ion  exchange chromatography on DEAE and cellulose and affinity  chromatography on heparin agarose. The effect of factor V activator on human factor V was studied by measuring the amidiolytic activity of the produced thrombin by activated factor V (Va).
Results: The results of SDS – PAGE  identified an activator  of   factor  V as a single  protein  band with a molecular  weight  of  29 KDa. This compound was converted, in the presence of calcium ions, to active factor Va. It also had arginine esterase  activity toward substrate BAEE ( -benzoyl arginine  ethyester) and a weak  amidase  activity  on  S-2222  (benzoyl- le-glu-Gly-Arg-p- nitroanilide).
Conclusion: The results of this study showed that the isolated factor is a specific activator on factor V.

Keywords


 
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