Specificity characterization of the alpha-mating factor hormone by Kex2 protease

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dc.citation.volume131
dc.contributor.authorManfredi, Marcella Araujo
dc.contributor.authorAntunes, Alyne Alexandrino
dc.contributor.authorPassos Jesus, Larissa de Oliveira
dc.contributor.authorJuliano, Maria Aparecida [UNIFESP]
dc.contributor.authorJuliano, Luiz [UNIFESP]
dc.contributor.authorde Souza Judice, Wagner Alves
dc.coverageParis
dc.date.accessioned2020-07-31T12:47:09Z
dc.date.available2020-07-31T12:47:09Z
dc.date.issued2016
dc.description.abstractKex2 is a Ca2+-dependent serine protease from S. cerevisiae. Characterization of the substrate specificity of Kex2 is of particular interest because this protease serves as the prototype of a large family of eukaryotic subtilisin-related proprotein-processing proteases that cleave sites consisting of pairs or clusters of basic residues. Our goal was to study the prime region subsite S' of Kex2 because previous studies have only taken into account non-prime sites using AMC substrates but not the specificity of prime sites identified through structural modeling or predicted cleavage sites. Therefore, we used peptides derived from Abz-KR down arrow EADQ-EDDnp and Abz-YKR down arrow EADQ-EDDnp based on the pro-a-mating factor sequence. The specificity of Kex2 due to basic residues at P-1' is affected by the type of residue in the P-3 position. Some residues in P-1' with large or bulky side chains yielded poor substrate specificity. The k(cat)/K-M values for peptides with P-2' substitutions containing Tyr in P-3 were higher than those obtained for the peptides without Tyr. In fact, P' and P modifications mainly promoted changes in k(cat) and K-M, respectively. The pH profile of Kex2 was fit to a double-sigmoidal pH-titration curve. The specificity results suggest that Kex2 might be involved in the processing of the putative cleavage sites in a poly peptide involved in cell elongation, hyphal formation and the processing of a toxin, which result in host cell lysis. In summary, the specificity of Kex2 is dependent on the set of interactions with prime and non-prime subsites, resulting in synergism. (C) 2016 Elsevier B.V. and Societe Francaise de Biochimie et Biologie Moleculaire (SFBBM). All rights reserved.en
dc.description.affiliationUniv Mogi das Cruzes UMC, Ctr Interdisciplinar Invest Bioquim, Mogi Das Cruzes, SP, Brazil
dc.description.affiliationUniv Fed Sao Paulo, Escola Paulista Med, Dept Biofis, Rua Thes Maio 100, BR-04044020 Sao Paulo, SP, Brazil
dc.description.affiliationUnifespDepartamento de Biofísica, Escola Paulista de Medicina, Universidade Federal de São Paulo, Rua Três de Maio, 100, São Paulo, SP, 04044-020, Brazil
dc.description.sourceWeb of Science
dc.description.sponsorshipFundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)
dc.description.sponsorshipIDFAPESP: 2014/02205-1
dc.format.extent149-158
dc.identifierhttp://dx.doi.org/10.1016/j.biochi.2016.10.003
dc.identifier.citationBiochimie. Paris, v. 131, p. 149-158, 2016.
dc.identifier.doi10.1016/j.biochi.2016.10.003
dc.identifier.issn0300-9084
dc.identifier.urihttps://repositorio.unifesp.br/handle/11600/56619
dc.identifier.wosWOS:000390513800015
dc.language.isoeng
dc.publisherElsevier France-Editions Scientifiques Medicales Elsevier
dc.relation.ispartofBiochimie
dc.rightsinfo:eu-repo/semantics/restrictedAccess
dc.subjectSpecificityen
dc.subjectKex2en
dc.subjectProprotein-processing proteasesen
dc.subjectalpha-mating factor hormoneen
dc.subjectSynergismen
dc.titleSpecificity characterization of the alpha-mating factor hormone by Kex2 proteaseen
dc.typeinfo:eu-repo/semantics/article
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