Catalytic properties of thimet oligopeptidase H600A mutant

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dc.contributor.author Machado, Mauricio F. M. [UNIFESP]
dc.contributor.author Marcondes, Marcelo F. [UNIFESP]
dc.contributor.author Rioli, Vanessa
dc.contributor.author Ferro, Emer S.
dc.contributor.author Juliano, Maria A. [UNIFESP]
dc.contributor.author Juliano, Luiz [UNIFESP]
dc.contributor.author Oliveira, Vitor [UNIFESP]
dc.date.accessioned 2016-01-24T13:59:35Z
dc.date.available 2016-01-24T13:59:35Z
dc.date.issued 2010-04-02
dc.identifier http://dx.doi.org/10.1016/j.bbrc.2010.03.045
dc.identifier.citation Biochemical and Biophysical Research Communications. San Diego: Academic Press Inc Elsevier Science, v. 394, n. 2, p. 429-433, 2010.
dc.identifier.issn 0006-291X
dc.identifier.uri http://repositorio.unifesp.br/handle/11600/32466
dc.description.abstract Thimet oligopeptidase (EC 3.4.24.15, TOP) is a metallo-oligopeptidase that participates in the intracellular metabolism of peptides. Predictions based on structurally analogous peptidases (Dcp and ACE-2) show that TOP can present a hinge-bend movement during substrate hydrolysis, what brings some residues closer to the substrate. One of these residues that in TOP crystallographic structure are far from the catalytic residues, but, moves toward the substrate considering this possible structural reorganization is His(600). in the present work, the role of His(600) of TOP was investigated by site-directed mutagenesis. TOP H600A mutant was characterized through analysis of S(1) and S(1)', specificity, pH-activity profile and inhibition by JA-2. Results showed that TOP His(600) residue makes important interactions with the substrate, supporting the prediction that His(600) moves toward the substrate due to a hinge movement similar to the Dcp and ACE-2. Furthermore, the mutation H600A affected both K(m) and k(cat), showing the importance of His(600) for both substrate binding and/or product release from active site. Changes in the pH-profile may indicate also the participation of His(600) in TOP catalysis, transferring a proton to the newly generated NH(2)-terminus or helping Tyr(605) and/or Tyr(612) in the intermediate oxyanion stabilization. (C) 2010 Elsevier Inc. All rights reserved. en
dc.description.sponsorship Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)
dc.description.sponsorship Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)
dc.format.extent 429-433
dc.language.iso eng
dc.publisher Elsevier B.V.
dc.relation.ispartof Biochemical and Biophysical Research Communications
dc.rights Acesso restrito
dc.subject EP24.15 en
dc.subject Substrate and inhibitor specificity en
dc.subject Site-directed mutagenesis en
dc.subject Thimet oligopeptidase en
dc.title Catalytic properties of thimet oligopeptidase H600A mutant en
dc.type Artigo
dc.rights.license http://www.elsevier.com/about/open-access/open-access-policies/article-posting-policy
dc.contributor.institution Universidade Federal de São Paulo (UNIFESP)
dc.contributor.institution Inst Butantan
dc.contributor.institution Universidade de São Paulo (USP)
dc.description.affiliation Universidade Federal de São Paulo, Dept Biophys, BR-04044020 São Paulo, Brazil
dc.description.affiliation Inst Butantan, Lab Especial Toxinol Aplicada, BR-05467010 São Paulo, Brazil
dc.description.affiliation Univ São Paulo, Dept Biol Celular & Desenvolvimento, BR-05508900 São Paulo, Brazil
dc.description.affiliationUnifesp Universidade Federal de São Paulo, Dept Biophys, BR-04044020 São Paulo, Brazil
dc.description.sponsorshipID FAPESP: 08/57336-2
dc.identifier.doi 10.1016/j.bbrc.2010.03.045
dc.description.source Web of Science
dc.identifier.wos WOS:000276732400033



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