Isolation and characterization of a serine proteinase with thrombin-like activity from the venom of the snake Bothrops asper

Brazilian Journal of Medical and Biological Research

A.v Pérez²  A Rucavado²  L Sanz¹  J.j Calvete¹  J.m Gutiérrez² 

[1] ,Universidad de Costa Rica Facultad de Microbiología Instituto Clodomiro PicadoSan José,Costa Rica

关键词: Snake venom;    Bothrops asper;    Serine proteinase;    Thrombin-like serine proteinase;    Defibrin(ogen)ation;   

DOI  :  10.1590/S0100-879X2006005000189

来源: SciELO

【 摘 要 】

A serine proteinase with thrombin-like activity was isolated from the venom of the Central American pit viper Bothrops asper. Isolation was performed by a combination of affinity chromatography on aminobenzamidine-Sepharose and ion-exchange chromatography on DEAE-Sepharose. The enzyme accounts for approximately 0.13% of the venom dry weight and has a molecular mass of 32 kDa as determined by SDS-PAGE, and of 27 kDa as determined by MALDI-TOF mass spectrometry. Its partial amino acid sequence shows high identity with snake venom serine proteinases and a complete identity with a cDNA clone previously sequenced from this species. The N-terminal sequence of the enzyme is VIGGDECNINEHRSLVVLFXSSGFL CAGTLVQDEWVLTAANCDSKNFQ. The enzyme induces clotting of plasma (minimum coagulant dose = 4.1 µg) and fibrinogen (minimum coagulant dose = 4.2 µg) in vitro, and promotes defibrin(ogen)ation in vivo (minimum defibrin(ogen)ating dose = 1.0 µg). In addition, when injected intravenously in mice at doses of 5 and 10 µg, it induces a series of behavioral changes, i.e., loss of the righting reflex, opisthotonus, and intermittent rotations over the long axis of the body, which closely resemble the `gyroxin-like' effect induced by other thrombin-like enzymes from snake venoms.

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