Purification and characterization of two arginine ester hydrolases from Vipera berus berus (common viper) venom

Abstract

M. Samel, E. Siigur and J. Siigur. Purification and characterization of two arginine ester hydrolases from Vipera berus berus (common viper) venom Toxicon 25, 379 – 388, 1987.—Two arginine ester hydrolases, designated EI and EII, consist of multiple molecular forms with pI values in the range 4.0–4.6 for EI and 3.3–3.9 for EII. Isoforms had identical molecular weights: 38,500 for EI and 41,000 for EII (SDS electrophoresis). The N-terminal amino acid for both enzymes was valine and their amino acid contents were very similar, with both containing carbohydrate. After treatment of EI and EII with neuraminidase both enzymes migrated identically in the electrofocusing system. Neither esterase hydrolyzed casein, α- N-benzoyl- dl-arginine- p-nitroanilide (BAPNA), yet both hydrolyzed α- N-benzoyl- l-arginine methylester (BAEE), p-tosyl- l-arginine methylester (TAME) and Pro-Phe-Arg-MCA. The esterase activities of the two enzymes were inhibited by organophosphorus inhibitors and benzamidine. The K m value for EI with BAEE was 3.3 × 10 −5M, with TAME 3.0 × 10 −5M, and for EII 2.7 × 10 −5M (BAEE) and 5.9 × 10 −5M (TAME). EII possessed kinin-releasing activity, as shown by the twitch response of an isolated rat uterus. The physiological role of EI is unknown. Neither esterase has thrombin-like or fibrinolytic activitives.