Controlled peptide solvation in portion-mixing libraries of FRET peptides: Improved specificity determination for dengue 2 virus NS2B-NS3 protease and human cathepsin S

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dc.contributor.authorAlves, Fabiana M.
dc.contributor.authorHirata, Izaura Y.
dc.contributor.authorGouvea, Iuri E.
dc.contributor.authorAlves, Marcio F. M.
dc.contributor.authorMeldal, Morten
dc.contributor.authorBromme, Dieter
dc.contributor.authorJuliano, Luiz
dc.contributor.authorJuliano, Maria A.
dc.contributor.institutionUniversidade Federal de São Paulo (UNIFESP)
dc.contributor.institutionCtr Gamle Carlsberg
dc.contributor.institutionUniv British Columbia
dc.date.accessioned2016-01-24T13:48:51Z
dc.date.available2016-01-24T13:48:51Z
dc.date.issued2007-07-01
dc.description.abstractThe solubility of peptides in aqueous buffers used for the enzyme assays is a common limitation for all peptide libraries. in principle, the more water-soluble peptides are, the more susceptible they will be to peptidase hydrolysis. We have demonstrated that this bias can be circumvented in a portion-mixing fluorescence resonance energy transfer (FRET) peptide library by introducing k (lysine in the D-form) in both termini of the peptides. This more solvated library and another one without the k were assayed using trypsin and chymotrypsin as standard peptidases with high selectivity for R and K and for hydrophobic F and Y, respectively. Significantly improved consistency of the information on substrate profiles was obtained from the solvated library. the influence of improved solvation on substrate specificity determination was successfully demonstrated by the difference in specificity observed between the two libraries employing the human cathepsin S (accepts acidic, basic, or neutral amino acids at P(1) position) and Dengue 2 virus NS2B-NS3 protease (high specificity to the pair of basic amino acids K-R, R-R, or Q-R/K at P(2)-P(1) positions). in conclusion, hydration of the peptides has a major influence on protease processing, and this bias can be reduced in bound peptide libraries, improving reliability.en
dc.description.affiliationUniversidade Federal de São Paulo, Escola Paulista Med, Dept Biophys, BR-04044020 São Paulo, Brazil
dc.description.affiliationCtr Gamle Carlsberg, SPOCC, Carlsberg Lab, DK-2500 Copenhagen, Denmark
dc.description.affiliationUniv British Columbia, Dept Dent, Vancouver, BC V6T 1Z3, Canada
dc.description.affiliationUniv British Columbia, UBC Ctr Blood Res, Vancouver, BC V6T 1Z3, Canada
dc.description.affiliationUnifespUniversidade Federal de São Paulo, Escola Paulista Med, Dept Biophys, BR-04044020 São Paulo, Brazil
dc.description.sourceWeb of Science
dc.format.extent627-634
dc.identifierhttp://dx.doi.org/10.1021/cc070042k
dc.identifier.citationJournal of Combinatorial Chemistry. Washington: Amer Chemical Soc, v. 9, n. 4, p. 627-634, 2007.
dc.identifier.doi10.1021/cc070042k
dc.identifier.issn1520-4766
dc.identifier.urihttp://repositorio.unifesp.br/handle/11600/29864
dc.identifier.wosWOS:000247820100011
dc.language.isoeng
dc.publisherAmer Chemical Soc
dc.relation.ispartofJournal of Combinatorial Chemistry
dc.rightsinfo:eu-repo/semantics/restrictedAccess
dc.titleControlled peptide solvation in portion-mixing libraries of FRET peptides: Improved specificity determination for dengue 2 virus NS2B-NS3 protease and human cathepsin Sen
dc.typeinfo:eu-repo/semantics/article
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