Purification and Partial Characterization of a Thrombin-Like Enzyme (AH۱۴۴) from Venom of Iranian Snake Agkistrodon Halys

The snake venom´s thrombin-like enzymes comprise a number of serine proteases, which are functionally and structurally related to thrombin. Purification and partial characterization of a thrombin-like enzyme from the venom of the Iranian snake, Agkistrodon halys, was the aim of this study. Purification was carried out by a combination of variety of chromatographic methods that included: gel filtration on Sephadex G-۵۰, ion-exchange chromatography on DEAE-Sepharose and HPLC with a C۱۸ column. A trial for the purification of protease resulted in an enzyme with specific activity of ۷۲۱.۲ (μmol/min/mg), which was purified by ۷۲.۱ fold. The purified thrombin-like enzyme designated AH۱۴۴ was found to have a molecular weight of approximately ۳۰.۵ kDa. This thrombin-like enzyme had the highest activity at ۳۷ °C and pH ۷.۵. Enzyme activity increased as its concentration increased, and the purified enzyme did not have any effect on casein. AH۱۴۴ demonstrated clotting and proteolytic activities in the presence of the human plasma and the synthetic substrate (BApNA), respectively. Data emphasized the possibility of AH۱۴۴ for quantitative determination of fibrinogen.

کلیدواژه ها:

Iranian snake venom ، Agkistrodon halys ، Thrombin-like enzyme ، Purification ، Coagulant activity