Respiratory response of phagocytes: terminal NADPH oxidase and the mechanisms of its activation.
The chemical composition, properties and activation mechanism of the O2(-)-forming NADPH oxidase of phagocytes were investigated, using partially purified enzyme preparations. Highly active NADPH oxidase was extracted as an aggregate of high Mr from the membranes of neutrophils and macrophages. The enzyme complex contained phospholipids and cytochrome b-245, very little FAD and almost no quinones or NAD(P)H-dye reductase activity. The purification of a polypeptide with a relative molecular mass of 31 500 strictly paralleled the purification of NADPH oxidase, suggesting that this polypeptide is a component of the enzyme. This protein was identified as cytochrome b -245 after dissociation of the proteolipid complex and purification of the cytochrome moiety. The 31 500 Mr protein was phosphorylated in enzyme preparations from activated but not from resting cells. The results indicate that: cytochrome b-245 is a major component of NADPH oxidase; the involvement of NAD(P)H dye reductases in the O2(-)-forming activity is questionable; the cytochrome b-245: FAD ratio in the enzyme complex is much higher than that indicated in crude preparations; the Mr of pig neutrophil cytochrome b-245 is 31 500; the activation of the O-2-forming system involves a process of phosphorylation of cytochrome b-245.
D000818:Animals; D003573:Cytochrome b Group / Q000378:metabolism; D004579:Electron Transport; D004789:Enzyme Activation; D005420:Flavoproteins / Q000378:metabolism; D006168:Guinea Pigs; D008264:Macrophages / Q000378:metabolism; D008956:Models, Chemical; D009247:NADH, NADPH Oxidoreductases / Q000302:isolation & purification / Q000378:metabolism*; D019255:NADPH Oxidases; D009504:Neutrophils / Q000378:metabolism; D010586:Phagocytes / Q000378:metabolism*; D010766:Phosphorylation; D011809:Quinones / Q000378:metabolism; D013481:Superoxides / Q000378:metabolism; D013552:Swine
D003573:Cytochrome b Group; D005420:Flavoproteins; D011809:Quinones; C035130:cytochrome b245; D013481:Superoxides; D009247:NADH, NADPH Oxidoreductases; D019255:NADPH Oxidases