Please use this identifier to cite or link to this item: http://hdl.handle.net/20.500.12779/4021
DC FieldValueLanguage
dc.contributor.authorDEL BELLO, Barbaraen_us
dc.contributor.authorMaellaro, Emiliaen_us
dc.contributor.authorSugherini, L.en_us
dc.contributor.authorSantucci, Annalisaen_us
dc.contributor.authorComporti, M.en_us
dc.contributor.authorCasini, A. F.en_us
dc.date.accessioned2021-03-30T14:41:10Z-
dc.date.available2021-03-30T14:41:10Z-
dc.date.issued1994-
dc.identifier.issn0264-6021en_US
dc.identifier.urihttp://hdl.handle.net/20.500.12779/4021-
dc.description25060en_US
dc.description.abstractRat liver cytosol has been found to reduce dehydroascorbic acid (DHAA) to ascorbic acid in the presence of NADPH. The enzyme responsible for such activity has been purified by ammonium sulphate fractionation, DEAE-Sepharose, Sephadex G-100 SF and Reactive Red column chromatography, with an overall recovery of 27%. SDS/PAGE of the purified enzyme showed one single protein band with an M(r) of 37,500. A similar value (36,800) was found by gel filtration on a Sephadex G-100 SF column. The results indicate that the enzyme is a homogeneous monomer. The Km for DHAA was 4.6 mM and the Vmax. was 1.55 units/mg of protein; for NADPH Km and Vmax. were 4.3 microM and 1.10 units/mg of protein respectively. The optimum pH was around 6.2. Several typical substrates and inhibitors of the aldo-keto reductase superfamily have been tested. The strong inhibition of DHAA reductase effected by steroidal and non-steroidal anti-inflammatory drugs, together with the ability to reduce 5 alpha-androstane-3,17-dione strongly, suggest the possibility that DHAA reductase corresponds to 3 alpha-hydroxysteroid dehydrogenase. Microsequence analysis performed on the electro-transferred enzyme band shows that the N-terminus is blocked. Internal primary structure data were obtained from CNBr-derived fragments and definitely proved the identity of NADPH-dependent DHAA reductase with 3 alpha-hydroxysteroid dehydrogenase.en_US
dc.language.isoenen_US
dc.relationNoneen_US
dc.relation.ispartofBIOCHEMICAL JOURNALen_US
dc.titlePurification of NADPH-dependent dehydroascorbate reductase from rat liver and its identificationen_US
dc.typeArticleen_US
dc.identifier.pmid7998972en_US
dc.identifier.scopus2-s2.0-0028007003en_US
dc.identifier.isiWOS:A1994PW11500011en_US
dc.identifier.urlhttp://www.biochemj.org/bj/304/0385/bj3040385.pdfen_US
dc.relation.volume304 (Pt 2)en_US
dc.description.firstpage385en_US
dc.description.lastpage390en_US
dc.description.thirdmissionNot applicableen_US
item.cerifentitytypePublications-
item.grantfulltextnone-
item.languageiso639-1en-
item.openairecristypehttp://purl.org/coar/resource_type/c_18cf-
item.openairetypeArticle-
item.fulltextNo Fulltext-
crisitem.author.orcid0000-0001-6976-9086-
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