The critical interaction of the metallopeptidase PHEX with heparan sulfate proteoglycans

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dc.contributor.author Barros, Nilana M. T. [UNIFESP]
dc.contributor.author Nascimento, Fabio D. [UNIFESP]
dc.contributor.author Oliveira, Vitor [UNIFESP]
dc.contributor.author Juliano, Maria Aparecida [UNIFESP]
dc.contributor.author Juliano, Luiz [UNIFESP]
dc.contributor.author Loisel, Thomas
dc.contributor.author Nader, Helena B. [UNIFESP]
dc.contributor.author Boileau, Guy
dc.contributor.author Tersariol, Ivarne L. S.
dc.contributor.author Carmona, Adriana K. [UNIFESP]
dc.date.accessioned 2016-01-24T13:49:22Z
dc.date.available 2016-01-24T13:49:22Z
dc.date.issued 2008-01-01
dc.identifier http://dx.doi.org/10.1016/j.biocel.2008.05.021
dc.identifier.citation International Journal of Biochemistry & Cell Biology. Oxford: Pergamon-Elsevier B.V., v. 40, n. 12, p. 2781-2792, 2008.
dc.identifier.issn 1357-2725
dc.identifier.uri http://repositorio.unifesp.br/handle/11600/30273
dc.description.abstract The PHEX gene (phosphate-regulating gene with homologies to endopeptidase on the X chromosome) identified as a mutated gene in patients with X-linked hypophosphatemia (XLH), encodes a protein (PHEX) that shows striking homologies to members of the M13 family of zinc metallopeptidases. in the present work the interaction of glycosaminoglycans with PHEX has been investigated by affinity chromatography, circular dichroism, protein intrinsic fluorescence analysis, hydrolysis of FRET substrates flow cytometry and confocal microscopy. PHEX was eluted from a heparin-Sepharose chromatography column at 0.8 M NaCl showing a strong interaction with heparin. Circular dichroism spectra and intrinsic fluorescence analysis showed that PHEX is protected by glycosaminoglycans against thermal denaturation. Heparin, heparan sulfate and chondroitin sulfate inhibited PHEX catalytic activity, however among them; heparin presented the highest inhibitory activity (K-i = 2.5 +/- 0.2 nM). Flow cytometry analysis showed that PHEX conjugated to Alexa Fluor 488 binds to the cell surface of CHO-K1, but did not bind to glycosaminoglycans defective cells CHO-745. Endogenous PHEX was detected at the cell surface of CHO-K1 colocalized with heparan sulfate proteoglycans, but was not found at the cell surface of glycosaminoglycans defective cells CHO-745. in permeabilized cells, PHEX was detected in endoplasmic reticulum of both cells. in addition, we observed that PHEX colocalizes with heparan sulfate at the cell surface of osteoblasts. This is the first report that the metallopeptidase PHEX is a heparin binding protein and that the interaction with GAGs modulates its enzymatic activity, protein stability and cellular trafficking. (c) 2008 Elsevier B.V. All rights reserved. en
dc.description.sponsorship Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)
dc.description.sponsorship Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)
dc.format.extent 2781-2792
dc.language.iso eng
dc.publisher Elsevier B.V.
dc.relation.ispartof International Journal of Biochemistry & Cell Biology
dc.rights Acesso restrito
dc.subject Phosphate-regulating gene with homologies to endopeptidase on the X chromosome (PHEX) en
dc.subject Metallopeptidase en
dc.subject Heparan sulfate en
dc.subject Heparin en
dc.subject Proteoglycans en
dc.title The critical interaction of the metallopeptidase PHEX with heparan sulfate proteoglycans en
dc.type Artigo
dc.rights.license http://www.elsevier.com/about/open-access/open-access-policies/article-posting-policy
dc.contributor.institution Universidade Federal de São Paulo (UNIFESP)
dc.contributor.institution Enobia Pharma Inc
dc.contributor.institution Univ Montreal
dc.contributor.institution Univ Mogi das Cruzes
dc.description.affiliation Universidade Federal de São Paulo, Dept Biofis, BR-04044020 São Paulo, Brazil
dc.description.affiliation Universidade Federal de São Paulo, Dept Bioquim, BR-04044020 São Paulo, Brazil
dc.description.affiliation Enobia Pharma Inc, Montreal, PQ H1W 4A4, Canada
dc.description.affiliation Univ Montreal, Dept Biochem, Montreal, PQ H3C 3J7, Canada
dc.description.affiliation Univ Mogi das Cruzes, CIIB, BR-08780210 Mogi Das Cruzes, SP, Brazil
dc.description.affiliationUnifesp Universidade Federal de São Paulo, Dept Biofis, BR-04044020 São Paulo, Brazil
dc.description.affiliationUnifesp Universidade Federal de São Paulo, Dept Bioquim, BR-04044020 São Paulo, Brazil
dc.identifier.doi 10.1016/j.biocel.2008.05.021
dc.description.source Web of Science
dc.identifier.wos WOS:000260125400014



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