2,6-Dichlorophenolindophenol-reducing activity of phagocytosis-associated NADPH oxidase.
The 2,6-dichlorophenolindophenol (DCIP)-reducing activity of the phagocytosis-associated NADPH oxidase was investigated using homogenates and a membrane fraction (F2) of elicited guinea pig peritoneal macrophages stimulated by phorbol myristate acetate. Essentially all of the stimulation-specific DCIP reduction under aerobic conditions could be inhibited when high concentrations of superoxide dismutase (SOD), about 10 times those usually used to inhibit the superoxide (O-2)-mediated cytochrome c reduction, were used. SOD inhibited the DCIP reduction by chemically generated O2- in the same manner as the stimulation-specific DCIP reduction by the macrophage F2, and the concentration of SOD necessary for 50% inhibition was about 10 times that for the reduction of cytochrome c. Under anaerobic conditions, however, the NADPH oxidase could reduce DCIP, though the rate was slow because we could not use a sufficiently high DCIP concentration. The observations indicate that the NADPH oxidase preferentially reduces oxygen under aerobic conditions, though the oxidase can reduce DCIP in the anaerobic state.
D015086:2,6-Dichloroindophenol / Q000378:metabolism*; D000693:Anaerobiosis; D000818:Animals; D003574:Cytochrome c Group / Q000378:metabolism; D006168:Guinea Pigs; D066298:In Vitro Techniques; D007215:Indophenol / Q000031:analogs & derivatives*; D008264:Macrophages / Q000201:enzymology; D009247:NADH, NADPH Oxidoreductases / Q000378:metabolism*; D019255:NADPH Oxidases; D010084:Oxidation-Reduction; D010587:Phagocytosis; D013482:Superoxide Dismutase / Q000378:metabolism
D003574:Cytochrome c Group; D007215:Indophenol; D015086:2,6-Dichloroindophenol; D013482:Superoxide Dismutase; D009247:NADH, NADPH Oxidoreductases; D019255:NADPH Oxidases