dc.contributor | Universidade Federal de São Paulo (UNIFESP) | |
dc.contributor | Univ Montreal | |
dc.creator | Barros, Nilana M. T. | |
dc.creator | Campos, Marcelo | |
dc.creator | Bersanetti, Patricia A. | |
dc.creator | Oliveira, Vitor | |
dc.creator | Juliano, Maria A. | |
dc.creator | Boileau, Guy | |
dc.creator | Juliano, Luiz | |
dc.creator | Carmona, Adriana K. | |
dc.date.accessioned | 2016-01-24T13:48:34Z | |
dc.date.accessioned | 2022-10-07T21:21:34Z | |
dc.date.available | 2016-01-24T13:48:34Z | |
dc.date.available | 2022-10-07T21:21:34Z | |
dc.date.created | 2016-01-24T13:48:34Z | |
dc.date.issued | 2007-04-01 | |
dc.identifier | Biological Chemistry. Berlin: Walter de Gruyter & Co, v. 388, n. 4, p. 447-455, 2007. | |
dc.identifier | 1431-6730 | |
dc.identifier | http://repositorio.unifesp.br/handle/11600/29657 | |
dc.identifier | 10.1515/BC.2007.048 | |
dc.identifier | WOS:000245407000011 | |
dc.identifier.uri | http://repositorioslatinoamericanos.uchile.cl/handle/2250/4028829 | |
dc.description.abstract | We examined the substrate specificity of the carboxydipeptidase activity of neprilysin (NEP) using fluorescence resonance energy transfer (FRET) peptides containing ortho-aminobenzoyl (Abz) and 2,4-dinitrophenyl (Dnp) as a donor/acceptor pair. Two peptide series with general sequences Abz-RXFK(Dnp)-OH and Abz-XRFK(Dnp)-OH (X denotes the position of the altered amino acid) were synthesized to study P, (cleavage at the X-F bond) and P-2 (cleavage at R-F bond) specificity, respectively. in these peptides a Phe residue was fixed in P-1' to fulfill the well-known NEP S-1' site requirement for a hydrophobic amino acid. in addition, we explored NEP capability to hydrolyze bradykinin (RPPGFSPFR) and its fluorescent derivative Abz-RPPGFSPFRQ-EDDnp (EDDnp 2,4-dinitrophenyl ethylenediamine). the enzyme acts upon bradykinin mainly as a carboxydipeptidase, preferentially cleaving Pro-Phe over the Gly-Phe bond in a 9:1 ratio, whereas Abz-RPPGFSPFRQ-EDDnp was hydrolyzed at the same bonds but at an inverted proportion of 1:9. the results show very efficient interaction of the substrates' C-terminal free carboxyl group with site S-2' of NEP, confirming the enzyme's preference to act as carboxydipeptidase at substrates with a free carboxyl-terminus. Using data gathered from our study, we developed sensitive and selective NEP substrates that permit continuous measurement of the enzyme activity, even in crude tissue extracts. | |
dc.language | eng | |
dc.publisher | Walter de Gruyter & Co | |
dc.relation | Biological Chemistry | |
dc.rights | Acesso restrito | |
dc.subject | bradykinin hydrolysis | |
dc.subject | carboxydipeptidase | |
dc.subject | continuous assay | |
dc.subject | FRET substrates | |
dc.subject | NEP | |
dc.subject | neprilysin | |
dc.title | Neprilysin carboxydipeptidase specificity studies and improvement in its detection with fluorescence energy transfer peptides | |
dc.type | Artigo | |