title: Regeneration of lipophilic antioxidants by NAD(P)H:quinone oxidoreductase 1.
abstract: The aim of this work was to study the activity of NAD(P)H:(quinone acceptor) oxidoreductase 1 (EC 1.6.99.2) in the regeneration of lipophilic antioxidants, alpha-tocopherol, and reduced-coenzyme Q analogs. First, we tested whether or not two isoforms of the NAD(P)H:(quinone acceptor) oxidoreductase 1 designated as "hydrophilic" and "hydrophobic" (H. J. Prochaska and P. Talalay, Journal of Biological Chemistry 261: 1372-1378, 1986) show differential enzyme activities towards hydrophilic or hydrophobic ubiquinone homologs. By chromatography on phenyl Sepharose, we purified the two isoforms from pig liver cytosol and measured their reduction of several ubiquinone homologs of different side chain length. We also studied by electron paramagnetic resonance the effect of NAD(P)H:(quinone acceptor) oxidoreductase 1 on steady-state levels of chromanoxyl radicals generated by linoleic acid and lipooxygenase and confirmed the enzyme's ability to protect alpha-tocopherol against oxidation induced with H(2)O(2)-Fe(2+). Our results demonstrated that the different hydrophobicities of the isoforms do not reflect different reactivities towards ubiquinones of different side chain length. In addition, electron paramagnetic resonance studies showed that in systems containing the reductase plus NADH, levels of chromanoxyl radicals were dramatically reduced. Morever, in the presence of oxidants, alpha-tocopherol was preserved by NAD(P)H:(quinone acceptor) oxidoreductase 1, supporting our hypothesis that regeneration of alpha-tocopherol may be one of the physiologic functions of this enzyme.
authors.affiliation: Departamento de Biología Celular, Fisiología e Inmunología, Universidad de Córdoba, Córdoba, Spain.
authors.forename: Rosario I
authors.initials: RI
authors.lastname: Bello
authors.forename: Valerian E
authors.initials: VE
authors.lastname: Kagan
authors.forename: Vladimir
authors.initials: V
authors.lastname: Tyurin
authors.forename: Francisco
authors.initials: F
authors.lastname: Navarro
authors.forename: Francisco J
authors.initials: FJ
authors.lastname: Alcaín
authors.forename: José M
authors.initials: JM
authors.lastname: Villalba
mesh_terms: D000818:Animals; D000975:Antioxidants / Q000378:metabolism*; D002384:Catalysis; D003600:Cytosol / Q000201:enzymology; D004578:Electron Spin Resonance Spectroscopy; D057927:Hydrophobic and Hydrophilic Interactions; D007527:Isoenzymes / Q000302:isolation & purification / Q000378:metabolism*; D008099:Liver / Q000201:enzymology*; D016660:NAD(P)H Dehydrogenase (Quinone) / Q000302:isolation & purification / Q000378:metabolism*; D009249:NADP / Q000378:metabolism; D010084:Oxidation-Reduction; D013552:Swine; D024502:alpha-Tocopherol / Q000378:metabolism
chemical_list: D000975:Antioxidants; D007527:Isoenzymes; D009249:NADP; D016660:NAD(P)H Dehydrogenase (Quinone); D024502:alpha-Tocopherol