Development of an operational substrate for ZapA, a metalloprotease secreted by the bacterium Proteus mirabilis

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dc.contributor.author Fernandes, B.l.
dc.contributor.author Anéas, M.a.f.
dc.contributor.author Juliano, Luiz [UNIFESP]
dc.contributor.author Palma, M.s.
dc.contributor.author Lebrun, Ivo
dc.contributor.author Portaro, Fernanda Calheta Vieira [UNIFESP]
dc.date.accessioned 2015-06-14T13:25:05Z
dc.date.available 2015-06-14T13:25:05Z
dc.date.issued 2000-07-01
dc.identifier http://dx.doi.org/10.1590/S0100-879X2000000700006
dc.identifier.citation Brazilian Journal of Medical and Biological Research. Associação Brasileira de Divulgação Científica, v. 33, n. 7, p. 765-770, 2000.
dc.identifier.issn 0100-879X
dc.identifier.uri http://repositorio.unifesp.br/handle/11600/992
dc.description.abstract The protease ZapA, secreted by Proteus mirabilis, has been considered to be a virulence factor of this opportunistic bacterium. The control of its expression requires the use of an appropriate methodology, which until now has not been developed. The present study focused on the replacement of azocasein with fluorogenic substrates, and on the definition of enzyme specificity. Eight fluorogenic substrates were tested, and the peptide Abz-Ala-Phe-Arg-Ser-Ala-Ala-Gln-EDDnp was found to be the most convenient for use as an operational substrate for ZapA. A single peptide bond (Arg-Ser) was cleaved with a Km of 4.6 µM, a k cat of 1.73 s-1, and a catalytic efficiency of 376 (mM s)-1. Another good substrate for ZapA was peptide 6 (Abz-Arg-Pro-Pro-Gly-Phe-Ser-Pro-Phe-Arg-Gln-EDDnp) which was cleaved at a single bond (Phe-Ser) with a Km of 13.6 µM, a k cat of 3.96 s-1 and a catalytic efficiency of 291 (mM s)-1. The properties of the amino acids flanking the scissile bonds were also evaluated, and no clear requirement for the amino acid residue at P1 was found, although the enzyme seems to have a preference for a hydrophobic residue at P2. en
dc.format.extent 765-770
dc.language.iso eng
dc.publisher Associação Brasileira de Divulgação Científica
dc.relation.ispartof Brazilian Journal of Medical and Biological Research
dc.rights Acesso aberto
dc.subject metalloprotease en
dc.subject substrate specificity en
dc.subject quenched fluorescence peptides en
dc.subject Proteus mirabilis en
dc.title Development of an operational substrate for ZapA, a metalloprotease secreted by the bacterium Proteus mirabilis en
dc.type Artigo
dc.contributor.institution Universidade de São Paulo (USP)
dc.contributor.institution Universidade Federal de São Paulo (UNIFESP)
dc.contributor.institution Universidade Estadual de São Paulo
dc.contributor.institution Instituto Butantan
dc.description.affiliation Universidade de São Paulo
dc.description.affiliation Universidade Federal de São Paulo (UNIFESP)
dc.description.affiliation Universidade Estadual de São Paulo
dc.description.affiliation Instituto Butantan
dc.description.affiliationUnifesp UNIFESP
dc.identifier.file S0100-879X2000000700006.pdf
dc.identifier.scielo S0100-879X2000000700006
dc.identifier.doi 10.1590/S0100-879X2000000700006
dc.description.source SciELO
dc.identifier.wos WOS:000088236100006



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