Reduction of ortho-aminobenzoyl-proline fluorescence and formation of pyrrolobenzodiazepine-5,11-dione

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dc.contributor.authorHirata, Izaura Yoshico [UNIFESP]
dc.contributor.authorCezari, Maria Helena Sedenho [UNIFESP]
dc.contributor.authorBoschcov, Paulo [UNIFESP]
dc.contributor.authorGarratt, Richard Charles
dc.contributor.authorOliva, Glaucius
dc.contributor.authorIto, Amando Siuiti
dc.contributor.authorSpisni, Alberto
dc.contributor.authorFranzoni, Lorella
dc.contributor.authorJuliano, Luiz [UNIFESP]
dc.contributor.institutionUniversidade Federal de São Paulo (UNIFESP)
dc.contributor.institutionUniversidade de São Paulo (USP)
dc.contributor.institutionUniv Parma
dc.date.accessioned2016-01-24T12:30:31Z
dc.date.available2016-01-24T12:30:31Z
dc.date.issued1998-01-01
dc.description.abstractThe ortho-aminobenzoic acid (Abz) group is widely employed as a fluorescent marker for peptides used as substrates for the study of proteolytic enzyme activity. in fact, a direct correlation has been observed between fluorescence intensity and enzyme activity. An unusual behavior of the fluorescence properties of this group, which would lead to erroneous evaluation of the enzyme activity, was observed when it is bound directly to proline. Here we report a systematic NMR, fluorescence and X-ray diffraction study of the compounds obtained from Boc-Abz-Pro-NH2, Boc-Abz-Pro-OH, as well as from various other Boc-Abz-Pro-X derivatives, after treatment with HCl or TFA under anhydrous conditions. We verified that, as recently reported, even under these synthetic conditions, deprotection of Boc-Abz-Pro-NH2 or Boc-Abz-Pro-OH leads to the formation of the same product: pyrrolobenzodiazepine-5,11-dione. However, the formation of this compound was not detected with Abz-Pro-N(CH3)(2), Abz-Pro-Leu-Gly-NH2 or Abz-pyrrolidine. for all these compounds we observed an unusual behavior for the fluorescence quantum yield of Abz that can be explained as the consequence of a non-radiative deactivation process produced, specifically, by the amidation of the Abz carboxyl group with proline or a similar secondary amine such as pyrrolidine. in conclusion, these results indicate that Abz cannot be used as an internal fluorescence marker for proteolytic enzyme activity when bound directly to proline.en
dc.description.affiliationUniversidade Federal de São Paulo, Escola Paulista Med, Dept Biophys, BR-04044020 São Paulo, Brazil
dc.description.affiliationUniv São Paulo, Inst Phys Sao Carlos, BR-13560250 Sao Carlos, SP, Brazil
dc.description.affiliationUniv São Paulo, Inst Phys, BR-01498970 São Paulo, Brazil
dc.description.affiliationUniv Parma, Inst Biol Chem, I-43100 Parma, Italy
dc.description.affiliationUnifespUniversidade Federal de São Paulo, Escola Paulista Med, Dept Biophys, BR-04044020 São Paulo, Brazil
dc.description.sourceWeb of Science
dc.format.extent19-28
dc.identifierhttp://dx.doi.org/10.1007/BF02443536
dc.identifier.citationLetters in Peptide Science. Dordrecht: Kluwer Academic Publ, v. 5, n. 1, p. 19-28, 1998.
dc.identifier.doi10.1007/BF02443536
dc.identifier.issn0929-5666
dc.identifier.urihttp://repositorio.unifesp.br/handle/11600/25838
dc.identifier.wosWOS:000073431500004
dc.language.isoeng
dc.publisherKluwer Academic Publ
dc.relation.ispartofLetters in Peptide Science
dc.rightsinfo:eu-repo/semantics/restrictedAccess
dc.subjectortho-aminobenzoyl-prolineen
dc.subjectpeptide synthesisen
dc.subjectprotease fluorescent substrateen
dc.subjectpyrrolobenzodiazepine-5,11-dioneen
dc.titleReduction of ortho-aminobenzoyl-proline fluorescence and formation of pyrrolobenzodiazepine-5,11-dioneen
dc.typeinfo:eu-repo/semantics/article
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