Isolation and Biochemical Characterization of a Non-Hemorrhagic Metalloproteasefrom Bothrops jararacussu Snake Venom. 

^1,2Marcussi, S.; ¹Fernandes, V. C.; ¹Silveira, L. B.; ¹Cambraia, R. S.; ³Mazzi, M. V.; ³Sant`Ana, C. D.; ¹França, S. C.; ³Sampaio, S. V.; ²Giglio, J. R. and ¹Soares, A. M.

1 Unidade de Biotecnologia-UNAERP, Ribeirão Preto-SP; 2 Depto Bioquímica e Imunologia, FMRP-USP, Ribeirão Preto-SP and 3 Depto. Análises Clínicas, Toxicológicas e Bromatológicas-FCFRP-USP, Ribeirão Preto-SP, Brasil.

Snake venom metalloproteases are responsible for several toxic and pharmacological effects, such as hemorrhagic, proteolytic, edema-inducing, dermonecrotic, bactericidal, anti-tumoral and thrombolytic. This work reports the isolation and enzymatic characterization of a new metalloproteases from B. jararacussu snake venom. The low molecular weight protease, BjussuMP-II, was isolated through two chromatographic steps, namely an ion-exchange on CM-Sepharose at pH 8.0, followed by a hydrophobic chromatography on Phenyl-Sepharose. Bjussu-MP-II showed a single polypeptide chain, Mr_˜22,000, by SDS-PAGE. This protein did not induce neither hemorrhage in vivo nor coagulation in vitro. It was however highly proteolytic upon fibrinogen, fibrin and casein. It induced dose-time-dependent proteolysis and was moderately stable under different temperatures and pH, it was also metal-dependent since EDTA completely inhibited its enzymatic activity. It also showed bactericidal activity against gram positive and gram negative. Ours results represent useful informations for the study of the action mechanism of low Mr metalloproteases from snake venoms and the participation of these enzymes in the envenomation process.

Financial support: FAPESP, CAPES, CNPq and UNAERP.

Correspondence to: smarcussi@rbi.fmrp.usp.br or andreimar@unaerp.br