Chromogenic and fluorogenic glycosylated and acetylglycosylated peptides as substrates for serine, thiol and aspartyl proteases

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dc.contributor.author Juliano, M. A.
dc.contributor.author Filira, F.
dc.contributor.author Gobbo, M.
dc.contributor.author Rocchi, R.
dc.contributor.author Del Nery, E.
dc.contributor.author Juliano, L.
dc.date.accessioned 2016-01-24T12:30:45Z
dc.date.available 2016-01-24T12:30:45Z
dc.date.issued 1999-02-01
dc.identifier http://dx.doi.org/10.1034/j.1399-3011.1999.00012.x
dc.identifier.citation Journal of Peptide Research. Copenhagen: Munksgaard Int Publ Ltd, v. 53, n. 2, p. 109-119, 1999.
dc.identifier.issn 1397-002X
dc.identifier.uri http://repositorio.unifesp.br/handle/11600/26031
dc.description.abstract We synthesized short chromogenic peptidyl-Arg-p-nitroanilides containing either (Gal beta)Ser or (Glc alpha,beta)Tyr at P-2 or P-3 sites as well as O-acetylated sugar moieties and studied their hydrolysis by bovine trypsin, papain, human tissue kallikrein and rat tonin. for comparison, the susceptibility to these enzymes of Acetyl-X-Arg-pNa and Acetyl-X-Phe-Arg-pNa series, in which X was Ala, Phe, Gin and Asn were examined. We also synthesized internally quenched fluorescent peptides with the amino acid sequence Phe(8)-His-Leu-Val-Ile-His-Asn(14) of hu man angiotensinogen, in which [GlcNAc beta]Asn was introduced before Phe(8) and/or after His(13) and ortho-aminobenzoic acid (Abz) and N-[2-, 4-dinitrophenyl]-ethylenediamine (EDDnp) were attached at N- and C-terminal ends as a donor/receptor fluorescent pair. These peptides were examined as substrates for human renin, human cathepsin D and porcine pepsin. the chromogenic substrates with hydrophilic sugar moiety increased their susceptibility to trypsin, tissue kallikrein and rat tonin. for papain, the effect of sugar depends on its position in the substrate, namely, at P-3 it is unfavorable, in contrast to the P-2 position that resulted in increasing affinity, as demonstrated by the higher inhibitory activity of Ac-(Gal beta)Ser-Arg-pNa in comparison to Ac-Ser-Arg-pNa, and by the hydrolysis of Ac-(Glc alpha,beta)Tyr-Arg-pNa. On the other hand, the acetylation of sugar hydroxyl groups improved hydrolysis of the susceptible peptides to all enzymes, except tonin. the P-4', glycosylated peptide [Abz-F-H-L-V-I-H-(GlcNAc beta)N-E-EDDnp], that corresponds to one of the natural glycosylation sites of angiotensinogen, was shown to be the only glycosylated substrate susceptible to human renin, and was hydrolysed with lower K-m and higher k(cat) values than the same peptide without the sugar moiety. Human cathepsin D and porcine pepsin are more tolerant to substrate glycosylation, hydrolysing both the P-4' and P-4 glycosylated substrates. en
dc.format.extent 109-119
dc.language.iso eng
dc.publisher Munksgaard Int Publ Ltd
dc.relation.ispartof Journal of Peptide Research
dc.rights Acesso restrito
dc.title Chromogenic and fluorogenic glycosylated and acetylglycosylated peptides as substrates for serine, thiol and aspartyl proteases en
dc.type Artigo
dc.contributor.institution Universidade Federal de São Paulo (UNIFESP)
dc.contributor.institution Univ Padua
dc.description.affiliation Escola Paulista Med, Dept Biofis, BR-04044020 São Paulo, Brazil
dc.description.affiliation Univ Padua, Dept Organ Chem, CNR, Biopolymer Res Ctr, I-35131 Padua, Italy
dc.description.affiliationUnifesp Escola Paulista Med, Dept Biofis, BR-04044020 São Paulo, Brazil
dc.identifier.doi 10.1034/j.1399-3011.1999.00012.x
dc.description.source Web of Science
dc.identifier.wos WOS:000079204500002



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