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Disulfide Biochemistry in 2-Cys Peroxiredoxin: Requirement of Glu50 and Arg146 for the Reduction of Yeast Tsa1 by Thioredoxin

dc.contributor.authorTairum, Carlos A. [UNESP]
dc.contributor.authorOliveira, Marcos A. de [UNESP]
dc.contributor.authorHorta, Bruno B.
dc.contributor.authorZara, Fernando Jose [UNESP]
dc.contributor.authorNetto, Luis E. S.
dc.contributor.institutionUniversidade Estadual Paulista (Unesp)
dc.contributor.institutionUniversidade de São Paulo (USP)
dc.date.accessioned2014-05-20T15:32:35Z
dc.date.available2014-05-20T15:32:35Z
dc.date.issued2012-11-23
dc.description.abstract2-Cys peroxiredoxin (Prx) enzymes are ubiquitously distributed peroxidases that make use of a peroxidatic cysteine (Cys(P)) to decompose hydroperoxides. A disulfide bond is generated as a consequence of the partial unfolding of the alpha-helix that contains Cys(P). Therefore, during its catalytic cycle, 2-Cys Prx alternates between two states, locally unfolded and fully folded. Tsa1 (thiol-specific antioxidant protein 1 from yeast) is by far the most abundant Cys-based peroxidase in Saccharomyces cerevisiae. In this work, we present the crystallographic structure at 2.8 angstrom resolution of Tsa1(C47S) in the decameric form [(alpha(2))(5)] with a DTT molecule bound to the active site, representing one of the few available reports of a 2-Cys Prx (AhpC-Prx1 subfamily) (AhpC, alkyl hydroperoxide reductase subunit C) structure that incorporates a ligand. The analysis of the Tsa1(C47S) structure indicated that G1u50 and Arg146 participate in the stabilization of the Cys(P) alpha-helix. As a consequence, we raised the hypothesis that G1u50 and Arg146 might be relevant to the Cys(P) reactivity. Therefore, Tsa1(E50A) and Tsa1(R146Q) mutants were generated and were still able to decompose hydrogen peroxide, presenting a second-order rate constant in the range of 10(6) M-1 S-1. Remarkably, although Tsa1(E50A) and Tsa1(R146Q) were efficiently reduced by the low-molecular-weight reductant DTT, these mutants displayed only marginal thioredoxin (Trx)-dependent peroxidase activity, indicating that G1u50 and Arg146 are important for the Tsa1-Trx interaction. These results may impact the comprehension of downstream events of signaling pathways that are triggered by the oxidation of critical Cys residues, such as Trx. (C) 2012 Elsevier Ltd. All rights reserved.en
dc.description.affiliationUniv Estadual Paulista, Dept Biol, BR-11330900 São Paulo, Brazil
dc.description.affiliationUniv São Paulo, Inst Biociencias, Dept Genet & Biol Evolut, BR-05508090 São Paulo, Brazil
dc.description.affiliationUniv Estadual Paulista, Dept Biol Aplicada Agr, BR-14884900 São Paulo, Brazil
dc.description.affiliationUnespUniv Estadual Paulista, Dept Biol, BR-11330900 São Paulo, Brazil
dc.description.affiliationUnespUniv Estadual Paulista, Dept Biol Aplicada Agr, BR-14884900 São Paulo, Brazil
dc.description.sponsorshipInstituto Nacional de Ciência e Tecnologia de Processos Redox em Biomedicina Redoxoma
dc.description.sponsorshipFundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)
dc.description.sponsorshipLaboratório Nacional de Luz Síncrotron (LNLS)
dc.description.sponsorshipIdINCT Processos Redox em Biomedicina Redoxoma: 2008/57721-3
dc.description.sponsorshipIdINCT Processos Redox em Biomedicina Redoxoma: 2008/573530
dc.description.sponsorshipIdFAPESP: 07/58147-6
dc.description.sponsorshipIdFAPESP: 07/50930-3
dc.description.sponsorshipIdBrazilian Synchrotron Light Laboratory (Laboratorio Nacional de Luz Sincrotron): D03B-CPR-1795
dc.format.extent28-41
dc.identifierhttp://dx.doi.org/10.1016/j.jmb.2012.09.008
dc.identifier.citationJournal of Molecular Biology. London: Academic Press Ltd- Elsevier B.V. Ltd, v. 424, n. 1-2, p. 28-41, 2012.
dc.identifier.doi10.1016/j.jmb.2012.09.008
dc.identifier.issn0022-2836
dc.identifier.urihttp://hdl.handle.net/11449/41445
dc.identifier.wosWOS:000312413100003
dc.language.isoeng
dc.publisherAcademic Press Ltd Elsevier B.V. Ltd
dc.relation.ispartofJournal of Molecular Biology
dc.relation.ispartofjcr4.894
dc.relation.ispartofsjr3,393
dc.rights.accessRightsAcesso restrito
dc.sourceWeb of Science
dc.subjectperoxiredoxinen
dc.subjectSaccharomyces cerevisiaeen
dc.subjectcrystal structureen
dc.subjectthioredoxinen
dc.subjectprotein-protein interactionsen
dc.titleDisulfide Biochemistry in 2-Cys Peroxiredoxin: Requirement of Glu50 and Arg146 for the Reduction of Yeast Tsa1 by Thioredoxinen
dc.typeArtigo
dcterms.licensehttp://www.elsevier.com/about/open-access/open-access-policies/article-posting-policy
dcterms.rightsHolderAcademic Press Ltd- Elsevier B.V. Ltd
dspace.entity.typePublication
unesp.campusUniversidade Estadual Paulista (UNESP), Faculdade de Ciências Agrárias e Veterinárias, Jaboticabalpt
unesp.departmentBiologia - FCAVpt

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