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dc.contributor.authorIerardi, D. F.
dc.contributor.authorPizauro, J. M.
dc.contributor.authorCiancaglini, P.
dc.identifier.citationBiochimica Et Biophysica Acta-biomembranes. Amsterdam: Elsevier B.V., v. 1567, n. 1-2, p. 183-192, 2002.
dc.description.abstractAlkaline phosphatase is required for the mineralization of bone and cartilage. This enzyme is localized in the matrix vesicle, which plays a role key in calcifying cartilage. In this paper we standardize a method to construction a resealed ghost cell-alkaline phosphatase system to mimic matrix vesicles and examine the kinetic behavior of the incorporated enzyme. Polidocanol-solubilized alkaline phosphatase, free of detergent, was incorporated into resealed ghost cells. This process was time-dependent and practically 50% of the enzyme was incorporated into the vesicles in 40 h of incubation, at 25 degreesC. Alkaline phosphatase-ghost cell systems were relatively homogeneous with diameters of about 300 nm and were more stable when stored at -20 degreesC.Alkaline phosphatase was completely released from the resealed ghost cell-system using only phospholipase C. These experiments confirm that the interaction between alkaline phosphatase and the lipid bilayer of resealed ghost cell is exclusively via glycosylphosphatidylinositol (GPI) anchor of the enzyme.An important point shown is that an enzyme bound to resealed ghost cell does not lose the ability to hydrolyze ATP, pyrophosphate and p-nitrophenyl phosphate (PNPP), but the presence of a ghost membrane, as a support of the enzyme, affects its kinetic properties. Moreover, calcium ions stimulate and phosphate ions inhibit the PNPPase activity of alkaline phosphatase present in resealed ghost cells. (C) 2002 Elsevier B.V. B.V. All rights reserved.en
dc.publisherElsevier B.V.
dc.relation.ispartofBiochimica et Biophysica Acta: Biomembranes
dc.sourceWeb of Science
dc.subjectAlkaline phosphatasept
dc.subjectOsseous platept
dc.subjectglycosylphosphatidylinositol (GPI) anchorpt
dc.subjectghost cellpt
dc.subjectendochondral ossificationpt
dc.titleErythrocyte ghost cell-alkaline phosphatase: construction and characterization of a vesicular system for use in biomineralization studiesen
dcterms.rightsHolderElsevier B.V.
dc.contributor.institutionUniversidade de São Paulo (USP)
dc.contributor.institutionUniversidade Estadual Paulista (Unesp)
dc.description.affiliationUSP, Fac Filosofia Ciências & Letras Ribeirao Pret, Dept Quim, BR-14040901 Ribeirao Preto, SP, Brazil
dc.description.affiliationUNESP, Fac Ciências Agr & Vet Jaboticabal, Dept Tecnol, BR-14884900 Jaboticabal, SP, Brazil
dc.description.affiliationUnespUNESP, Fac Ciências Agr & Vet Jaboticabal, Dept Tecnol, BR-14884900 Jaboticabal, SP, Brazil
dc.rights.accessRightsAcesso restrito
unesp.campusUniversidade Estadual Paulista (Unesp), Faculdade de Ciências Agrárias e Veterinárias, Jaboticabalpt[3][2]
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