Effect of a protrombin activator isolated from Iranian Echis carinatus venom on hemostasis

Echis carinatus venom is a complex mixture of toxins. This venom contains metalloproteinases which convert prothrombin to meizothrombin. The prothrombin activator leads to the formation of small blood clots inside the blood vessels throughout the body. To understand the effect mechanism of Iranian Echis carinatus venom, in this study we investigated the effect of EV on human plasma proteins (prothrombin and fibrinogen) and on blood coagulation. The aim was the purification and characterization of procoagulant factor from the Iranian Echis Carinatus venom and the evaluation of the procoagulant activity on human plasma.Crude venom from the Iranian snake species E. carinatus was selected. The prothrombin activator was purified from the crude venom of Echis carinatus by combination of the procedures by gel filtration and ion-exchange chromatography. The sample was applied into a Superdex G-75 column and fractions were collected at 4°C and their absorbances were recorded at 280 nm. Than sample was loaded into ion exchange column (DEAE-Sepharose) and equilibrated with 50 mM Tris-HCl buffer, pH 8.2 and eluted with a liner gradient of Nacl concentration from 0.0 to 0.5 mM. The peaks were monitored at 280 nm. The Iranian E. carinatus venom was able to coagulate human plasma very rapidly. The coagulation time was reduced from 13.4 seconds (SD= ±0.59) to 8.6 seconds (SD= ±0.64) when human plasma was treated with crude venom (concentraion of venom was 1 mg/ml).