Isolation and biochemical characterization of a fibrinolytic proteinase from Bothrops leucurus (white-tailed jararaca) snake venom

Isolation and biochemical characterization of a fibrinolytic proteinase from Bothrops leucurus (white-tailed jararaca) snake venom

Autor Bello, C. A. Google Scholar
Hermogenes, ALN Google Scholar
Magalhaes, A. Google Scholar
Veiga, S. S. Google Scholar
Gremski, L. H. Google Scholar
Richardson, M. Google Scholar
Sanchez, E. F. Google Scholar
Instituição Ezequiel Dias Fdn
Univ Fed Parana
Universidade Federal de São Paulo (UNIFESP)
Resumo In investigations aimed at characterizing snake venom clot-dissolving enzymes, we have purified a fibrinolytic proteinase from the venom of Bothrops leucurus (white-tailed-jararaca). the proteinase was purified to homogeneity by a combination of molecular sieve chromatography on Sephacryl S-200 and ion-exchange chromatography oil CM Sepharose. the enzyme called leucrolysin-a (leuc-a), is a 23 kDa metalloendopeptidase since it is inhibited by EDTA. PMSF, a specific serine proteinase inhibitor had no effect on leuc-a activity. the amino acid sequence was established by Edman degradation of overlapping peptides generated by a variety of selective cleavage procedures. Leuc-a is related in amino acid sequence to reprolysins. the protein is composed of 200 amino acid residues in a single polypeptide chain, possessing a blocked NH2-terminus and containing no carbohydrate. the proteinase showed proteolytic activity on dimethylcasein and on fibrin (specific activity = 21.6 units/mg and 17.5 units/Pg, respectively; Crude venom = 8.0 units/mg and 9.5 units/mu g). Leuc-a degrades fibrin and fibrinogen by hydrolysis of the alpha chains. Moreover, the enzyme was capable of cleaving plasma fibronectin but not the basement membrane protein laminin. Leuc-a. cleaved the Ala(14)-Leu(15) and Tyr(16)-Leu(17) bonds in oxidized insulin B chain. the pH optimum of the proteolysis of dimethylcasein by leuc-a was about pH 7.0. Antibody raised in rabbit against the Purified enzyme reacted with leuc-a and with the crude venom of B. leucurus. in vitro studies revealed that leuc-a dissolves clots made either from Purified fibrinogen or from whole blood, and unlike some other venom fibrinolytic metallopeptidases, leuc-a is devoid of hemorrhagic activity when injected (up to 100 mu g) Subcutaneously into mice. (c) 2005 Elsevier SAS. All rights reserved.
Palavra-chave leucurolysin
metalloproteinase
fibrinolysis
Bothrops leucurus
snake venoms
Idioma Inglês
Data de publicação 2006-02-01
Publicado em Biochimie. Paris: Elsevier France-editions Scientifiques Medicales Elsevier, v. 88, n. 2, p. 189-200, 2006.
ISSN 0300-9084 (Sherpa/Romeo, fator de impacto)
Publicador Elsevier B.V.
Extensão 189-200
Fonte http://dx.doi.org/10.1016/j.biochi.2005.07.008
Direito de acesso Acesso restrito
Tipo Artigo
Web of Science WOS:000234879900009
Endereço permanente http://repositorio.unifesp.br/handle/11600/28721

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