Please use this identifier to cite or link to this item: https://repositorio.unifesp.br/handle/11600/28443
Title: Characterization of thimet- and neurolysin-like activities in Escherichia coli M3A peptidases and description of a specific substrate
Authors: Paschoalin, T.
Carmona, A. K.
Oliveira, V
Juliano, L.
Travassos, L. R.
Universidade Federal de São Paulo (UNIFESP)
Keywords: thimet oligopeptidase
neurolysin
oligopeptidase A (OpdA)
dipeptidyl dipeptidase (Dcp)
Escherichia coli
expression vector
recombinant protein
co-purification
M3A subfamily
specific substrate
Issue Date: 1-Sep-2005
Publisher: Elsevier B.V.
Citation: Archives of Biochemistry and Biophysics. New York: Elsevier B.V., v. 441, n. 1, p. 25-34, 2005.
Abstract: 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.
URI: http://repositorio.unifesp.br/handle/11600/28443
ISSN: 0003-9861
Other Identifiers: http://dx.doi.org/10.1016/j.abb.2005.06.011
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