title: Hydrogen-peroxide-induced heme degradation in red blood cells: the protective roles of catalase and glutathione peroxidase.
abstract: Catalase and glutathione peroxidase (GSHPX) react with red cell hydrogen peroxide. A number of recent studies indicate that catalase is the primary enzyme responsible for protecting the red cell from hydrogen peroxide. We have used flow cytometry in intact cells as a sensitive measure of the hydrogen-peroxide-induced formation of fluorescent heme degradation products. Using this method, we have been able to delineate a unique role for GSHPX in protecting the red cell from hydrogen peroxide. For extracellular hydrogen peroxide, catalase completely protected the cells, while the ability of GSHPX to protect the cells was limited by the availability of glutathione. The effect of endogenously generated hydrogen peroxide in conjunction with hemoglobin autoxidation was investigated by in vitro incubation studies. These studies indicate that fluorescent products are not formed during incubation unless the glutathione is reduced to at least 40% of its initial value as a result of incubation or by reacting the glutathione with iodoacetamide. Reactive catalase only slows down the depletion of glutathione, but does not directly prevent the formation of these fluorescent products. The unique role of GSHPX is attributed to its ability to react with hydrogen peroxide generated in close proximity to the red cell membrane in conjunction with the autoxidation of membrane-bound hemoglobin.
authors.affiliation: Molecular Dynamics Section, Gerontology Research Center, National Institute on Aging, National Institutes of Health, 5600 Nathan Shock Drive, Baltimore, MD 21224, USA.
authors.forename: Enika
authors.initials: E
authors.lastname: Nagababu
authors.forename: Francis J
authors.initials: FJ
authors.lastname: Chrest
authors.forename: Joseph M
authors.initials: JM
authors.lastname: Rifkind
mesh_terms: D002374:Catalase / Q000037:antagonists & inhibitors / Q000378:metabolism*; D004305:Dose-Response Relationship, Drug; D004905:Erythrocyte Aging; D004910:Erythrocyte Membrane / Q000378:metabolism; D004912:Erythrocytes / Q000201:enzymology / Q000378:metabolism*; D005296:Ferrous Compounds; D005434:Flow Cytometry; D005453:Fluorescence; D005979:Glutathione Peroxidase / Q000037:antagonists & inhibitors / Q000378:metabolism*; D006418:Heme / Q000737:chemistry / Q000378:metabolism*; D006801:Humans; D006861:Hydrogen Peroxide / Q000378:metabolism* / Q000494:pharmacology; D007460:Iodoacetamide; D015227:Lipid Peroxidation; D009573:Nitrites; D010084:Oxidation-Reduction; D018384:Oxidative Stress; D013696:Temperature; D013997:Time Factors
chemical_list: D005296:Ferrous Compounds; D009573:Nitrites; D006418:Heme; D006861:Hydrogen Peroxide; D002374:Catalase; D005979:Glutathione Peroxidase; D007460:Iodoacetamide