Mechanism of action and determination of the best substrate for a thrombin-like enzyme from Lachesis muta muta venom by regression analysis of the kinetic parameters determined with peptidyl p-nitroanilide substrates

Mechanism of action and determination of the best substrate for a thrombin-like enzyme from Lachesis muta muta venom by regression analysis of the kinetic parameters determined with peptidyl p-nitroanilide substrates

Autor Magalhaes, HPB Google Scholar
Magalhaes, A. Google Scholar
Juliano, L. Google Scholar
Nelson, D. L. Google Scholar
Rogana, E. Google Scholar
Instituição Universidade Federal de Minas Gerais (UFMG)
Fudacao Ezequiel Dias MG
Universidade Federal de São Paulo (UNIFESP)
Resumo The kinetic behavior of a thrombin-like enzyme from Lachesis muta muta venom has been studied with 13 tripeptidyl p-nitroanilide substrates. Eight substrates were unprotected at the N terminus and were used for the regression analysis of the experimentally determined kinetic parameters 1/K-m, k(cat), and k(cat)/K-m. the individual contribution of each amino acid side chain to the kinetic parameters was calculated. the amino acid sequence of the ideal Substrate (D-Pro-Leu-Arg-pNA) was determined from a regression analysis for each kinetic parameter. This result was confirmed experimentally. the structural analysis of the tripeptides showed that the binding to the S-3 sub-site had a small effect on K-m. the binding of L-Leu to the S-2 sub-site increased k(cat) without changing the value of K-m. the analysis of the kinetic parameters revealed that, in the binding of L-Leu to the S-2 subsite, the enzyme bound the transition state configuration of the substrate/product trans formation more tightly than that of the substrate. (c) 2006 Elsevier B.V. All rights reserved.
Palavra-chave thrombin-like enzyme
Lachesis muta muta
tripeptidyl p-nitroanilides
Idioma Inglês
Data de publicação 2006-03-15
Publicado em Toxicon. Oxford: Pergamon-Elsevier B.V., v. 47, n. 4, p. 453-458, 2006.
ISSN 0041-0101 (Sherpa/Romeo, fator de impacto)
Publicador Elsevier B.V.
Extensão 453-458
Fonte http://dx.doi.org/10.1016/j.toxicon.2006.01.001
Direito de acesso Acesso restrito
Tipo Artigo
Web of Science WOS:000236534700010
Endereço permanente http://repositorio.unifesp.br/handle/11600/28792

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