A prothrombin activator from Bothrops erythromelas (jararaca-da-seca) snake venom: characterization and molecular cloning

A prothrombin activator from Bothrops erythromelas (jararaca-da-seca) snake venom: characterization and molecular cloning

Author Silva, Márcia B. Google Scholar
Schattner, Mirta Google Scholar
Ramos, Celso RR Google Scholar
Junqueira-de-Azevedo, Inácio LM Google Scholar
Guarnieri, Míriam C. Google Scholar
Lazzari, Maria A. Google Scholar
Sampaio, Claudio Augusto Machado Autor UNIFESP Google Scholar
Pozner, Roberto G. Google Scholar
Ventura, Janaína S. Google Scholar
Ho, Paulo L. Google Scholar
Chudzinski-Tavassi, Ana M. Google Scholar
Institution Inst Butantan
Universidade Federal de Pernambuco (UFPE)
Consejo Nacl Invest Cient & Tecn
Universidade Federal de São Paulo (UNIFESP)
Abstract A novel prothrombin activator enzyme, which we have named 'berythractivase', was isolated from Bothrops erythromelas (jararaca-da-seca) snake venom. Berythractivase was purified by a single cation-exchange-chromatography step on a Resource S (Amersham Biosciences) column. the overall purification (31-fold) indicates that berythractivase comprises about 5% of the crude venom. It is a single-chain protein with a molecular mass of 78 kDa. SDS/PAGE of prothrombin after activation by berythractivase showed fragment patterns similar to those generated by group A prothrombin activators, which convert prothrombin into meizothrombin, independent of the prothrombinase complex. Chelating agents, such as EDTA and o-phenanthroline, rapidly inhibited the enzymic activity of berythractivase, like a typical metalloproteinase. Human fibrinogen Aalpha-chain was slowly digested only. after longer incubation with berythractivase, and no effect on the beta- or gamma-chains was observed. Berythractivase was also capable of triggering endothelial proinflammatory and procoagulant cell responses. von Willebrand factor-was released, and the surface expression of both intracellular adhesion molecule-1 and E-selectin was up-regulated by berythractivase in cultured human umbilical-vein endothelial cells. the complete berythractivase cDNA was cloned from a B. erythromelas venom-gland cDNA library. the cDNA sequence possesses 2330 bp and encodes a preproprotein with significant sequence similarity to many other mature metalloproteinases reported from snake venoms. Berythractivase contains metalloproteinase, desintegrin-like and cysteine-rich domains. However, berythractivase did not elicit any haemorrhagic response. These results show that, although the primary structure of berythractivase is related to that of snake-venom haemorrhagic metalloproteinases and functionally similar to group A prothrombin activators, it is a prothrombin activator devoid of haemorrhagic activity. This is a feature not observed for most of the snake venom metalloproteinases, including the group A prothrombin activators.
Keywords berythractivase
coagulation
haemostasis
metalloproteinases
thrombosis
Language English
Date 2003-01-01
Published in Biochemical Journal. London: Portland Press, v. 369, p. 129-139, 2003.
ISSN 0264-6021 (Sherpa/Romeo, impact factor)
Publisher Portland Press
Extent 129-139
Origin http://dx.doi.org/10.1042/BJ20020449
Access rights Open access Open Access
Type Article
Web of Science ID WOS:000180871000014
URI http://repositorio.unifesp.br/handle/11600/27076

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