Characterization of thimet- and neurolysin-like activities in Escherichia coli M3A peptidases and description of a specific substrate

Characterization of thimet- and neurolysin-like activities in Escherichia coli M3A peptidases and description of a specific substrate

Autor Paschoalin, T. Google Scholar
Carmona, A. K. Google Scholar
Oliveira, V Google Scholar
Juliano, L. Google Scholar
Travassos, L. R. Google Scholar
Instituição Universidade Federal de São Paulo (UNIFESP)
Resumo M3A oligopeptidases from Escherichia coli, with hydrolytic properties similar to Zn-dependent mammalian thimet oligopeptidase (EP 24.15) and neurolysin (EP 24.16), were studied aiming at identification of comparative enzyme and substrate specificity, hydrolytic products, and susceptibility to inhibitors. Fluorescent peptides, neurotensin (NT) and bradykinin (BK), were used as substrates for bacterial lysates. Bacterial enzymes were totally inhibited by o-phenanthrolin, JA-2 and partially by Pro-Ile, but not by leupeptin, PMSF, E-64, and Z-Pro-Prolinal, using internally quenched Abz-GFSPFRQ-EDDnp as substrate. the molecular mass of the bacterial oligopeptidase activity (77-78 kDa) was determined by gel filtration, and the effect of inhibitors, including captopril, suggested that it results from a combination of oligopeptidase A (OpdA) and peptidyl dipeptidase Dcp (77.1 and 77.5 kDa, respectively). Recombinant OpdA cloned from the same E. coli strain entirely reproduced the primary cleavage of fluorescent peptides, NT and BK, by the bacterial lysate. Genes encoding these M3A enzymes were those recognized in E. coli genome, bearing identity at the amino acid level (25-31%) with mammalian Zn-dependent oligopeptidases. We also describe a substrate, Abz-GFSPFRQ-EDDnp, that differentiates bacterial and mammalian oligopeptidases. (C) 2005 Elsevier Inc. All rights reserved.
Palavra-chave thimet oligopeptidase
neurolysin
oligopeptidase A (OpdA)
dipeptidyl dipeptidase (Dcp)
Escherichia coli
expression vector
recombinant protein
co-purification
M3A subfamily
specific substrate
Idioma Inglês
Data de publicação 2005-09-01
Publicado em Archives of Biochemistry and Biophysics. New York: Elsevier B.V., v. 441, n. 1, p. 25-34, 2005.
ISSN 0003-9861 (Sherpa/Romeo, fator de impacto)
Publicador Elsevier B.V.
Extensão 25-34
Fonte http://dx.doi.org/10.1016/j.abb.2005.06.011
Direito de acesso Acesso restrito
Tipo Artigo
Web of Science WOS:000231678600003
Endereço permanente http://repositorio.unifesp.br/handle/11600/28443

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