Human recombinant endopeptidase PHEX has a strict S-1 ' specificity for acidic residues and cleaves peptides derived from fibroblast growth factor-23 and matrix extracellular phosphoglycoprotein

Página do item simplificado
dc.contributor.authorCampos, Marcelo [UNIFESP]
dc.contributor.authorCouture, Constance
dc.contributor.authorHirata, Izaura Y. [UNIFESP]
dc.contributor.authorJuliano, Maria Aparecida [UNIFESP]
dc.contributor.authorLoisel, Thomas P.
dc.contributor.authorCrine, Philippe
dc.contributor.authorJuliano, Luiz [UNIFESP]
dc.contributor.authorBoileau, Guy
dc.contributor.authorCarmona, Adriana Karaoglanovic [UNIFESP]
dc.contributor.institutionUniversidade Federal de São Paulo (UNIFESP)
dc.contributor.institutionUniv Montreal
dc.contributor.institutionBioMep Inc
dc.date.accessioned2016-01-24T12:33:55Z
dc.date.available2016-01-24T12:33:55Z
dc.date.issued2003-07-01
dc.description.abstractThe PHEX gene (phosphate-regulating gene with homologies to endopeptidases on the X chromosome) encodes a protein (PHEX) with structural homologies to members of the M13 family of zinc metallo-endopeptidases. Mutations in the PHEX gene are responsible for X-linked hypophosphataemia in humans. However, the mechanism by which loss of PHEX function results in the disease phenotype, and the endogenous PHEX substrate(s) remain unknown. in order to study PHEX substrate specificity, combinatorial fluorescent-quenched peptide libraries containing o-aminobenzoic acid (Abz) and 2,4-dinitrophenyl (Dnp) as the donor-acceptor pair were synthesized and tested as PHEX substrates. PHEX showed a strict requirement for acidic amino acid residues (aspartate or glutamate) in S-1' subsite, with a strong preference for aspartate. Subsites S-2', S-1 and S-2 exhibited less defined specificity requirements, but the presence of leucine, proline or glycine in P-2', or valine, isoleucine or histidine in P, precluded hydrolysis of the substrate by the enzyme. the peptide Abz-GFSDYK(Dnp)-OH, which contains the most favourable residues in the P-2 to P-2' positions, was hydrolysed by PHEX at the N-terminus of aspartate with a k(cat)/K-m of 167 mM(-1) . s(-1). in addition, using quenched fluorescence peptides derived from fibroblast growth factor-23 and matrix extracellular phosphoglycoprotein sequences flanked by Abz and N-(2,4-dinitrophenyl)ethylenediamine, we showed that these physiologically relevant proteins are potential PHEX substrates. Finally, our results clearly indicate that PHEX does not have neprilysin-like substrate specificity.en
dc.description.affiliationUniversidade Federal de São Paulo, Dept Biophys, Escola Paulista Med, BR-04044020 São Paulo, Brazil
dc.description.affiliationUniv Montreal, Dept Biochem, Montreal, PQ H3C 3J7, Canada
dc.description.affiliationBioMep Inc, Montreal, PQ H4B 2L5, Canada
dc.description.affiliationUnifespUniversidade Federal de São Paulo, Dept Biophys, Escola Paulista Med, BR-04044020 São Paulo, Brazil
dc.description.sourceWeb of Science
dc.format.extent271-279
dc.identifierhttp://dx.doi.org/10.1042/BJ20030287
dc.identifier.citationBiochemical Journal. London: Portland Press, v. 373, p. 271-279, 2003.
dc.identifier.doi10.1042/BJ20030287
dc.identifier.issn0264-6021
dc.identifier.urihttp://repositorio.unifesp.br/handle/11600/27296
dc.identifier.wosWOS:000184060200028
dc.language.isoeng
dc.publisherPortland Press
dc.relation.ispartofBiochemical Journal
dc.rightsinfo:eu-repo/semantics/openAccess
dc.subjectcombinatorial libraryen
dc.subjectinternally quenched fluorogenic substrateen
dc.subjectM13 endopeptidaseen
dc.subjectPHEX substrate specificityen
dc.titleHuman recombinant endopeptidase PHEX has a strict S-1 ' specificity for acidic residues and cleaves peptides derived from fibroblast growth factor-23 and matrix extracellular phosphoglycoproteinen
dc.typeinfo:eu-repo/semantics/article
Arquivos
Coleções
Em verificação - Geral