Osseous plate alkaline phosphatase is anchored by GPI.
dc.contributor.author | Pizauro, J. M. [UNESP] | |
dc.contributor.author | Ciancaglini, P. [UNESP] | |
dc.contributor.author | Leone, F. A. [UNESP] | |
dc.contributor.institution | Universidade Estadual Paulista (UNESP) | |
dc.date.accessioned | 2022-04-28T19:54:02Z | |
dc.date.available | 2022-04-28T19:54:02Z | |
dc.date.issued | 1994-01-01 | |
dc.description.abstract | Alkaline phosphatase activity was released up to 100% from the membrane by using 0.1 U of phosphatidylinositol-specific phospholipase C from B. thuringiensis. The M(r) of solubilized enzyme was 145,000 by Sephacryl S-300 gel filtration and 66,000 by SDS-PAGE, suggesting a dimeric structure. Solubilization of the membrane-bound enzyme with phospholipase C did not destroy its ability to hydrolyze p-nitrophenyl phosphate (PNPP) (264.3 mumol min-1 mg-1),ATP (42.0 mumol min-1 mg-1) and pyrophosphate (28.4 mumol min-1 mg-1). The hydrolysis of ATP and PNPP by solubilized enzyme exhibited Michaelian kinetics with K0.5 = 70 and 979 microM, respectively. For pyrophosphate, K0.5 was 128 microM and site-site interactions were observed (n = 1.4). Magnesium ions were stimulatory (Kd = 1.5 mM) but zinc ions were powerful non-competitive inhibitors (Kd = 6.2 microM) of solubilized enzyme. Treatment of solubilized alkaline phosphatase with Chellex 100 reduced the original PNPPase activity to 5%. Cobalt (K0.5 = 10.1 microM), magnesium (K0.5 = 29.5 microM) and manganese ions (K0.5 = 5 microM) restored the activity of the apoenzyme with positive cooperativity, suggesting that phosphatidylinositol-specific phospholipase C-solubilized alkaline phosphatase is a metalloenzyme. The stimulation of the apoenzyme by calcium ions (K0.5 = 653 microM) was lower than that observed for the other ions (26%) and exhibited site-site interactions (n = 0.7). Zinc ions had no effect on the apoenzyme of the solubilized enzyme. | en |
dc.description.affiliation | Departamento de Technologia Faculdade de Ciências Agrárias e Veterinárias de Jaboticabal UNESP | |
dc.description.affiliationUnesp | Departamento de Technologia Faculdade de Ciências Agrárias e Veterinárias de Jaboticabal UNESP | |
dc.format.extent | 453-456 | |
dc.identifier.citation | Brazilian journal of medical and biological research = Revista brasileira de pesquisas médicas e biológicas / Sociedade Brasileira de Biofísica ... [et al.], v. 27, n. 2, p. 453-456, 1994. | |
dc.identifier.issn | 0100-879X | |
dc.identifier.scopus | 2-s2.0-0028379425 | |
dc.identifier.uri | http://hdl.handle.net/11449/223982 | |
dc.language.iso | eng | |
dc.relation.ispartof | Brazilian journal of medical and biological research = Revista brasileira de pesquisas médicas e biológicas / Sociedade Brasileira de Biofísica ... [et al.] | |
dc.source | Scopus | |
dc.title | Osseous plate alkaline phosphatase is anchored by GPI. | en |
dc.type | Artigo | |
unesp.department | Tecnologia - FCAV | pt |