Foot and mouth disease leader protease (Lb(pro)): Investigation of prime side specificity allows the synthesis of a potent inhibitor

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dc.contributor.authorNogueira Santos, Jorge Alexandre
dc.contributor.authorAssis, Diego M.
dc.contributor.authorGouvea, Iuri Estrada
dc.contributor.authorJudice, Wagner A. S.
dc.contributor.authorIzidoro, Mario Augusto
dc.contributor.authorJuliano, Maria Aparecida
dc.contributor.authorSkern, Tim
dc.contributor.authorJuliano, Luiz [UNIFESP]
dc.contributor.institutionUniversidade Federal de São Paulo (UNIFESP)
dc.contributor.institutionUniv Mogi das Cruzes
dc.contributor.institutionMed Univ Vienna
dc.date.accessioned2016-01-24T14:26:54Z
dc.date.available2016-01-24T14:26:54Z
dc.date.issued2012-03-01
dc.description.abstractFoot and mouth disease virus expresses its genetic information as a single polyprotein that is translated from the single-stranded RNA genome. Proteinases contained within the polyprotein then generate the mature viral proteins. the leader protease (Lb(pro)) performs the initial cleavage by freeing itself from the growing polypeptide chain; subsequently, Lb(pro) cleaves the two homologues of the host cell protein eukaryotic initiation factor 4G (eIF4G). We showed that Lb(pro) possesses specific binding sites at the non prime side from S-1 down to S-7 [Santos et al. (2009) Biochemistry, 48, 7948-7958]. Here, we demonstrate that Lb(pro) has high prime side specificity at least down to the S-5' site. Lb(pro) is thus not only one of the smallest papain-like cysteine peptidases but also one of the most specific. It can still however cleave between both K down arrow G and G down arrow R pairs. We further determined the two-step irreversible inhibition (E + I <-> EI -> E - I) kinetic parameters of two known irreversible epoxide-based inhibitors of cysteine proteinases, E64 and CA074 on Lb(pro) that show for the reversible step (E + I <-> EI) K-i = 3.4 mu M and 11.6 mu M, and for the irreversible step (EI -> E-I) k(4) = 0.16 and 0.06 min(-1), respectively. Knowledge of the Lb(pro) specificity led us to extend E64 by addition of the dipeptide R-P. This compound, termed E64-R-P-NH2, irreversibly inhibited Lb(pro) with a K-i = 30 nM and k(4) = 0.01 min(-1) and can serve as the basis for design of specific inhibitors of FMDV replication. (C) 2011 Elsevier Masson SAS. All rights reserved.en
dc.description.affiliationUniversidade Federal de São Paulo, Dept Biophys, Escola Paulista Med, UNIFESP, BR-04044020 São Paulo, Brazil
dc.description.affiliationUniv Mogi das Cruzes, Ctr Interdisciplinar Invest Bioquim, BR-08780911 Mogi Das Cruzes, Brazil
dc.description.affiliationMed Univ Vienna, Max F Perutz Labs, A-1030 Vienna, Austria
dc.description.affiliationUnifespUniversidade Federal de São Paulo, Dept Biophys, Escola Paulista Med, UNIFESP, BR-04044020 São Paulo, Brazil
dc.description.sourceWeb of Science
dc.description.sponsorshipFundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)
dc.description.sponsorshipConselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)
dc.description.sponsorshipInstituto Nacional de Ciencia e Tecnologia em Fluidos Complexos (INCT-Fx) in Brazil
dc.description.sponsorshipAustrian Science Foundation
dc.description.sponsorshipIDAustrian Science Foundation: P20889
dc.format.extent711-718
dc.identifierhttp://dx.doi.org/10.1016/j.biochi.2011.10.016
dc.identifier.citationBiochimie. Paris: Elsevier France-editions Scientifiques Medicales Elsevier, v. 94, n. 3, p. 711-718, 2012.
dc.identifier.doi10.1016/j.biochi.2011.10.016
dc.identifier.issn0300-9084
dc.identifier.urihttp://repositorio.unifesp.br/handle/11600/34671
dc.identifier.wosWOS:000301332200015
dc.language.isoeng
dc.publisherElsevier B.V.
dc.relation.ispartofBiochimie
dc.rightsAcesso restrito
dc.rights.licensehttp://www.elsevier.com/about/open-access/open-access-policies/article-posting-policy
dc.subjectCysteine proteaseen
dc.subjectFluorescent peptidesen
dc.subjectCathepsinen
dc.subjectPicornavirusen
dc.subjectProtease inhibitorsen
dc.titleFoot and mouth disease leader protease (Lb(pro)): Investigation of prime side specificity allows the synthesis of a potent inhibitoren
dc.typeArtigo
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