Positional-scanning combinatorial libraries of fluorescence resonance energy transfer peptides to define substrate specificity of carboxydipeptidases: assays with human cathepsin B

dc.contributor.authorCotrin, S. S.
dc.contributor.authorPuzer, L.
dc.contributor.authorJudice, WAD
dc.contributor.authorJuliano, L.
dc.contributor.authorCarmona, A. K.
dc.contributor.authorJuliano, M. A.
dc.contributor.institutionUniversidade Federal de São Paulo (UNIFESP)
dc.date.accessioned2016-01-24T12:37:31Z
dc.date.available2016-01-24T12:37:31Z
dc.date.issued2004-12-15
dc.description.abstractWe have developed positional scanning synthetic combinatorial libraries to define the substrate specificity of carboxydipeptidases. the library Abz-GXXZXK(Dnp)-OH, where Abz is ortho-aminobenzoic acid, K(Dnp) is N-c-2,4-dinitrophenyl-lysine with free carboxyl group, the Z position was successively occupied with 1 of 19 amino acids (eysteine was omitted), and X represents randomly incorporated residues, was assayed initially with human cathepsin B, and arginine was defined as one of the best residues at the P, position. To examine the selectivity of S-1(1) S-2, and S-3 subsites, the sublibraries Abz-GXXRZK(Dnp)-OH, AbzGXZRXK(Dnp)-OH, and Abz-GZXRXK(Dnp)-OH were then synthesized. the peptide Abz-GIVRAK(Dnp)-OH, which contains the most favorable residues in the P-3-P-1, positions identified by screening of the libraries with cathepsin B, was hydrolyzed by this enzyme with k(cat)/K-m = 7288 mM(-1) s(-1). This peptide is the most efficient substrate described for cathepsin B to this point, and it is highly selective for the enzyme among the lysosomal cysteine proteases. (C) 2004 Elsevier Inc. All rights reserved.en
dc.description.affiliationUNIFESP, Escola Paulista Med, Dept Biophys, BR-04044020 São Paulo, Brazil
dc.description.affiliationUnifespUNIFESP, Escola Paulista Med, Dept Biophys, BR-04044020 São Paulo, Brazil
dc.description.sourceWeb of Science
dc.format.extent244-252
dc.identifierhttp://dx.doi.org/10.1016/j.ab.2004.09.012
dc.identifier.citationAnalytical Biochemistry. San Diego: Academic Press Inc Elsevier Science, v. 335, n. 2, p. 244-252, 2004.
dc.identifier.doi10.1016/j.ab.2004.09.012
dc.identifier.issn0003-2697
dc.identifier.urihttp://repositorio.unifesp.br/handle/11600/28050
dc.identifier.wosWOS:000225502800009
dc.language.isoeng
dc.publisherElsevier B.V.
dc.relation.ispartofAnalytical Biochemistry
dc.rightsAcesso restrito
dc.rights.licensehttp://www.elsevier.com/about/open-access/open-access-policies/article-posting-policy
dc.subjectcysteine peptidaseen
dc.subjectlysosomeen
dc.subjectamino acidsen
dc.subjectexopeptidaseen
dc.titlePositional-scanning combinatorial libraries of fluorescence resonance energy transfer peptides to define substrate specificity of carboxydipeptidases: assays with human cathepsin Ben
dc.typeArtigo
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