The cytosolic activating factors p47phox and p67phox have distinct roles in the regulation of electron flow in NADPH oxidase.
We have previously shown that the human neutrophil superoxide-generating NADPH oxidase possesses a novel dye reductase activity (Cross, A.R., Yarchover, J. L., and Curnutte, J.T. (1994) J. Biol. Chem. 269, 21448-21454). This activity exhibited an absolute requirement for the cytosolic activating factor p67phox but not for p47phox, suggesting that p67phox and p47phox have individual roles in controlling electron flow from NADPH to oxygen. Here, we provide direct evidence that p67phox alone can facilitate electron flow from NADPH to the flavin center of NADPH oxidase in the absence of p47phox, resulting in the reduction of enzyme FAD, whereas the presence of p47phox is required in order for electron transfer to proceed beyond the flavin center to the heme in cytochrome b-245 and thence to oxygen.
Cross
A R
AR
Department of Molecular and Experimental Medicine, Scripps Research Institute, La Jolla, California 92037.
Curnutte
J T
JT
D005415:Flavins / Q000378:metabolism; D005690:Galactose / Q000494:pharmacology; D005692:Galactose Oxidase / Q000494:pharmacology; D006418:Heme / Q000378:metabolism; D006801:Humans; D009247:NADH, NADPH Oxidoreductases / Q000737:chemistry*; D009252:NADPH Dehydrogenase / Q000502:physiology*; D019255:NADPH Oxidases; D009504:Neutrophils / Q000201:enzymology*; D010084:Oxidation-Reduction; D010750:Phosphoproteins / Q000502:physiology*
D005415:Flavins; D010750:Phosphoproteins; C079789:neutrophil cytosol factor 67K; D006418:Heme; D005692:Galactose Oxidase; D009247:NADH, NADPH Oxidoreductases; D019255:NADPH Oxidases; C060505:neutrophil cytosolic factor 1; D009252:NADPH Dehydrogenase; D005690:Galactose