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dc.contributor.authorSabbagh, Yves
dc.contributor.authorBoileau, Guy
dc.contributor.authorCampos, Marcelo [UNIFESP]
dc.contributor.authorCarmona, Adriana Karaoglanovic [UNIFESP]
dc.contributor.authorTenenhouse, Harriet S.
dc.date.accessioned2016-01-24T12:33:48Z
dc.date.available2016-01-24T12:33:48Z
dc.date.issued2003-05-01
dc.identifierhttp://dx.doi.org/10.1210/jc.2002-021809
dc.identifier.citationJournal of Clinical Endocrinology & Metabolism. Chevy Chase: Endocrine Soc, v. 88, n. 5, p. 2213-2222, 2003.
dc.identifier.issn0021-972X
dc.identifier.urihttp://repositorio.unifesp.br/handle/11600/27223
dc.description.abstractThe PHEX gene that is mutated in patients with X-linked hypophosphatemia (XLH) encodes a protein homologous to the M13 family of zinc metallopeptidases. the present study was undertaken to assess the impact of nine PHEX missense mutations on cellular trafficking, endopeptidase activity, and protein conformation. Secreted forms of wild-type and mutant PHEX proteins were generated by PCR mutagenesis; these included C85R, D237G, Y317F, G579R, G579V, S711R, A720T, and F731Y identified in XLH patients, and E581V, which in neutral endopeptidase 24.11 abolishes catalytic activity but not plasma membrane localization. the wild-type and D237G, Y317F, E581V, and F731Y proteins were terminally glycosylated and secreted into the medium, whereas the C85R, G579R, G579V, S711R, and A720T proteins were trapped inside the transfected cells. Growing the cells at 26 C permitted the secretion of G579V, S711R, and A720T proteins, although the yield of rescued G579V was insufficient for further analysis. Endopeptidase activity of secreted and rescued PHEX proteins, assessed using a novel internally quenched fluorogenic peptide substrate, revealed that E581V and S711R are completely inactive; D237G and Y317F exhibit 50-60% of wild-type activity; and A720T and F731Y retain full catalytic activity. Conformational analysis by limited proteolysis demonstrated that F731Y is more sensitive to trypsin and D237G is more resistant to endoproteinase Glu-c than the wild-type protein. Thus, defects in protein trafficking, endopeptidase activity, and protein conformation account for loss of PHEX function in XLH patients harboring these missense mutations.en
dc.format.extent2213-2222
dc.language.isoeng
dc.publisherEndocrine Soc
dc.relation.ispartofJournal of Clinical Endocrinology & Metabolism
dc.rightsAcesso aberto
dc.titleStructure and function of disease-causing missense mutations in the PHEX geneen
dc.typeArtigo
dc.contributor.institutionMcGill Univ
dc.contributor.institutionUniv Montreal
dc.contributor.institutionUniversidade Federal de São Paulo (UNIFESP)
dc.description.affiliationMcGill Univ, Montreal Childrens Hosp, Res Inst, Montreal, PQ H3Z 2Z3, Canada
dc.description.affiliationMcGill Univ, Dept Biol, Montreal, PQ H3Z 2Z3, Canada
dc.description.affiliationMcGill Univ, Dept Pediat, Montreal, PQ H3Z 2Z3, Canada
dc.description.affiliationMcGill Univ, Dept Human Genet, Montreal, PQ H3Z 2Z3, Canada
dc.description.affiliationUniv Montreal, Dept Biochem, Montreal, PQ H3C 3J7, Canada
dc.description.affiliationUniversidade Federal de São Paulo, Dept Biophys, Escola Paulista Med, BR-04044020 São Paulo, Brazil
dc.description.affiliationUnifespUniversidade Federal de São Paulo, Dept Biophys, Escola Paulista Med, BR-04044020 São Paulo, Brazil
dc.identifier.doi10.1210/jc.2002-021809
dc.description.sourceWeb of Science
dc.identifier.wosWOS:000182595500045


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