Characterization of two cysteine proteinases secreted by Fasciola hepatica and demonstration of their kininogenase activity

Characterization of two cysteine proteinases secreted by Fasciola hepatica and demonstration of their kininogenase activity

Autor Cordova, M. Google Scholar
Jara, J. Google Scholar
Del Nery, E. Google Scholar
Hirata, I. Y. Google Scholar
Araujo, M. S. Google Scholar
Carmona, A. K. Google Scholar
Juliano, M. A. Google Scholar
Juliano, L. Google Scholar
Instituição Universidade Federal de São Paulo (UNIFESP)
Univ Peruana Cayetano Heredia
Resumo We have isolated and purified two cysteine proteinases of molecular weights 25 and 26 kDa, secreted by Fasciola hepatica adult worm. Their 15 N-terminal residues were found to be identical to those of earlier described cathepsin L-like enzymes, isolated from the same source, reported as CL1 and CL2. Radioimmunoassay experiments have shown that these CL1- (25 kDa) and CL2-like (26 kDa) cysteine proteinases mediated kinin release from high molecular weight kininogen (HMWK). Lys-bradykinin (KRPPGFSPFR) was characterized as the kinin released from a synthetic fragment of HMWK from Leu(373) to Ile(393) (Abz-LGMISLMKRPPGFSPFRSSRI-NH2) labeled with the fluorescent group Abz (ortho-aminobenzoic acid). We examined the activity of CL1- and CL2-like on internally quenched fluorescent peptides containing HMWK sequences, in which Met(379)-Lys(380) or Arg(389)-Ser(390) bonds were present in the. middle of the molecules. These peptides. were flanked by the fluorescent donor-acceptor pair Abz and EDDnp (N-[2,4-dinitrophenyl] ethylenediamine). Peptidyl-methylcoumarin amides (MCA) were used to study the substrate specificity requirements. the enzymes presented significantly lower K-m values at pH 8.0. the inverse was observed with the k(cat) values, which were higher at pH 5.0. (C) 2001 Elsevier Science B.V. All rights reserved.
Palavra-chave cathepsin L-like
Fasciola hepatica cysteine proteinases
kinin-releasing activity
Idioma Inglês
Data de publicação 2001-09-03
Publicado em Molecular and Biochemical Parasitology. Amsterdam: Elsevier B.V., v. 116, n. 2, p. 109-115, 2001.
ISSN 0166-6851 (Sherpa/Romeo, fator de impacto)
Publicador Elsevier B.V.
Extensão 109-115
Fonte http://dx.doi.org/10.1016/S0166-6851(01)00309-7
Direito de acesso Acesso restrito
Tipo Artigo
Web of Science WOS:000171004600001
Endereço permanente http://repositorio.unifesp.br/handle/11600/26626

Exibir registro completo




Arquivo

Arquivo Tamanho Formato Visualização

Não existem arquivos associados a este item.

Este item está nas seguintes coleções

Buscar


Navegar

Minha conta