title: Coisolation of glutathione peroxidase, catalase and superoxide dismutase from human erythrocytes.
abstract: Glutathione peroxidase (GSH-Px; glutathione: hydrogen peroxide oxidoreductase; EC 1.11.1.9), catalase (H2O2: H2O2 oxidoreductase; EC 1.11.1.6) and superoxide dismutase (superoxide: superoxide oxidoreductase; EC 1.15.1.1) were coisolated from human erythrocyte lysate by chromatography on DEAE-cellulose. Glutathione peroxidase was separated from superoxide dismutase and catalase by thiol-disulfide exchange chromatography and then purified to approximately 90% homogeneity by gel permeation chromatography and dye-ligand affinity chromatography. Catalase and superoxide dismutase were separated from each other and purified further by gel permeation chromatography. Catalase was then purified to approximately 90% homogeneity by ammonium sulfate precipitation and superoxide dismutase was purified to apparent homogeneity by hydrophobic interaction chromatography. The results for glutathione peroxidase represent an improvement of approximately 10-fold in yield and 3-fold in specific activity compared with the established method for the purification of this enzyme. The yields for superoxide dismutase and catalase were high (45 mg and 232 mg, respectively, from 820 ml of washed packed cells), and the specific activities of both enzymes were comparable to values found in the literature.
authors.affiliation: Medical Biophysics Branch, Whiteshell Nuclear Research Establishment, Atomic Energy of Canada Ltd., Pinawa, Manitoba.
authors.forename: T M
authors.initials: TM
authors.lastname: Stepanik
authors.forename: D D
authors.initials: DD
authors.lastname: Ewing
mesh_terms: D002374:Catalase / Q000302:isolation & purification*; D002846:Chromatography, Affinity; D002850:Chromatography, Gel; D002851:Chromatography, High Pressure Liquid; D002852:Chromatography, Ion Exchange; D004912:Erythrocytes / Q000201:enzymology*; D005979:Glutathione Peroxidase / Q000302:isolation & purification*; D006801:Humans; D013482:Superoxide Dismutase / Q000302:isolation & purification*
chemical_list: D002374:Catalase; D005979:Glutathione Peroxidase; D013482:Superoxide Dismutase