title: Nitric oxide prevents oxidative damage produced by tert-butyl hydroperoxide in erythroleukemia cells via nitrosylation of heme and non-heme iron. Electron paramagnetic resonance evidence.
abstract: We studied protective effects of NO against tert-butylhydroperoxide (t-BuOOH)-induced oxidations in a subline of human erythroleukemia K562 cells with different intracellular hemoglobin (Hb) concentrations. t-BuOOH-induced formation of oxoferryl-Hb-derived free radical species in cells was demonstrated by low temperature EPR spectroscopy. Intensity of the signals was proportional to Hb concentrations and was correlated with cell viability. Peroxidation of phosphatidylcholine, phosphatidylethanolamine, phosphatidylserine, phosphatidylinositol, and cardiolipin metabolically labeled with oxidation-sensitive cis-parinaric acid was induced by t-BuOOH. An NO donor, (Z)-1-[N-(3-ammoniopropyl)-N-(n-propyl)amino]-diazen-1-iu m-1, 2-diolate], produced non-heme iron dinitrosyl complexes and hexa- and pentacoordinated Hb-nitrosyl complexes in the cells. Nitrosylation of non-heme iron centers and Hb-heme protected against t-BuOOH-induced: (a) formation of oxoferryl-Hb-derived free radical species, (b) peroxidation of cis-parinaric acid-labeled phospholipids, and (c) cytotoxicity. Since NO did not inhibit peroxidation induced by an azo-initiator of peroxyl radicals, 2, 2'-azobis(2,4-dimethylvaleronitrile), protective effects of NO were due to formation of iron-nitrosyl complexes whose redox interactions with t-BuOOH prevented generation of oxoferryl-Hb-derived free radical species.
authors.affiliation: Department of Respiratory Research, Division of Medicine, Walter Reed Army Institute of Research, Washington, D. C. 20307, USA.
authors.forename: N V
authors.initials: NV
authors.lastname: Gorbunov
authors.forename: J C
authors.initials: JC
authors.lastname: Yalowich
authors.forename: A
authors.initials: A
authors.lastname: Gaddam
authors.forename: P
authors.initials: P
authors.lastname: Thampatty
authors.forename: V B
authors.initials: VB
authors.lastname: Ritov
authors.forename: E R
authors.initials: ER
authors.lastname: Kisin
authors.forename: N M
authors.initials: NM
authors.lastname: Elsayed
authors.forename: V E
authors.initials: VE
authors.lastname: Kagan
mesh_terms: D002470:Cell Survival; D004578:Electron Spin Resonance Spectroscopy; D005609:Free Radicals; D006418:Heme / Q000378:metabolism*; D006454:Hemoglobins / Q000378:metabolism; D006801:Humans; D007700:Kinetics; D004915:Leukemia, Erythroblastic, Acute / Q000378:metabolism*; D009569:Nitric Oxide / Q000494:pharmacology*; D018384:Oxidative Stress; D010545:Peroxides / Q000494:pharmacology*; D017382:Reactive Oxygen Species / Q000378:metabolism*; D014407:Tumor Cells, Cultured; D020122:tert-Butylhydroperoxide
chemical_list: D005609:Free Radicals; D006454:Hemoglobins; D010545:Peroxides; D017382:Reactive Oxygen Species; D009569:Nitric Oxide; D006418:Heme; D020122:tert-Butylhydroperoxide