Substrate specificity of recombinant dengue 2 virus NS2B-NS3 protease: Influence of natural and unnatural basic amino acids on hydrolysis of synthetic fluorescent substrates

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dc.contributor.authorGouvea, I. E.
dc.contributor.authorIzidoro, M. A.
dc.contributor.authorJudice, W. A. S.
dc.contributor.authorCezari, M. H. S.
dc.contributor.authorCaliendo, G.
dc.contributor.authorSantagada, V.
dc.contributor.authorSantos, C. N. D. dos
dc.contributor.authorQueiroz, M. H.
dc.contributor.authorJuliano, M. A.
dc.contributor.authorYoung, P. R.
dc.contributor.authorFairlie, D. P.
dc.contributor.authorJuliano, L.
dc.contributor.institutionUniversidade Federal de São Paulo (UNIFESP)
dc.contributor.institutionUniv Naples Federico II
dc.contributor.institutionInst Biol Mol Parana
dc.contributor.institutionUniv Queensland
dc.date.accessioned2016-01-24T12:41:49Z
dc.date.available2016-01-24T12:41:49Z
dc.date.issued2007-01-15
dc.description.abstractA recombinant dengue 2 virus NS2B-NS3 protease (NS means non-structural virus protein) was compared with human furin for the capacity to process short peptide substrates corresponding to seven native Substrate cleavage sites in the dengue viral polyprotein. Using fluorescence resonance energy transfer peptides to measure kinetics, the processing of these substrates was found to be selective for the Dengue protease. Substrates containing two or three basic amino acids (Arg or Lys) in tandem were found to be the best, with Abz-AKRRSQ-EDDnp being the most efficiently cleaved. the hydrolysis of dipeptide substrates Bz-X-Arg-MCA where X is a non-natural basic amino acid were also kinetically examined, the best substrates containing aliphatic basic amino acids. Our results indicated that proteolytic processing by dengue NS3 protease, tethered to its activating NS2B co-factor, was strongly inhibited by Ca2+ and kosmotropic salts of the Hofmeister's series, and significantly influenced by substrate modifications between S-4 and S'. Incorporation of basic non-natural amino acids in short peptide substrates had significant but differential effects on K-m and k(cat) suggesting that further dissection of their influences on substrate affinity might enable the development of effective dengue protease inhibitors. (c) 2006 Elsevier Inc. All rights reserved.en
dc.description.affiliationUniversidade Federal de São Paulo, Escola Paulista Med, Dept Biofis, BR-04044020 São Paulo, Brazil
dc.description.affiliationUniv Naples Federico II, Dipartimento Chim Farmaceut & Tossicol, I-80131 Naples, Italy
dc.description.affiliationInst Biol Mol Parana, BR-81350010 Curitiba, Parana, Brazil
dc.description.affiliationUniv Queensland, Sch Mol & Microbial Sci, St Lucia, Qld 4072, Australia
dc.description.affiliationUniv Queensland, Inst Mol Biosci, St Lucia, Qld 4072, Australia
dc.description.affiliationUnifespUniversidade Federal de São Paulo, Escola Paulista Med, Dept Biofis, BR-04044020 São Paulo, Brazil
dc.description.sourceWeb of Science
dc.format.extent187-196
dc.identifierhttp://dx.doi.org/10.1016/j.abb.2006.11.005
dc.identifier.citationArchives of Biochemistry and Biophysics. New York: Elsevier B.V., v. 457, n. 2, p. 187-196, 2007.
dc.identifier.doi10.1016/j.abb.2006.11.005
dc.identifier.issn0003-9861
dc.identifier.urihttp://repositorio.unifesp.br/handle/11600/29459
dc.identifier.wosWOS:000243572800008
dc.language.isoeng
dc.publisherElsevier B.V.
dc.relation.ispartofArchives of Biochemistry and Biophysics
dc.rightsinfo:eu-repo/semantics/restrictedAccess
dc.rights.licensehttp://www.elsevier.com/about/open-access/open-access-policies/article-posting-policy
dc.subjectproteaseen
dc.subjectpeptidesen
dc.subjectdengueen
dc.titleSubstrate specificity of recombinant dengue 2 virus NS2B-NS3 protease: Influence of natural and unnatural basic amino acids on hydrolysis of synthetic fluorescent substratesen
dc.typeinfo:eu-repo/semantics/article
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