The reduction of cytochrome b558 and the activity of the respiratory burst oxidase from human neutrophils.
It is known that in respiratory burst oxidase preparations engaged in O2- production, cytochrome b558, a characteristic oxidase component, is partly reduced. This result has been interpreted in terms of a mechanism in which cytochrome b558 functions as an electron-carrying component of the respiratory burst oxidase, its level of reduction reflecting a steady-state partitioning of the cytochrome between reduced and oxidized forms as it ferries electrons from NADPH to oxygen. Kinetic arguments based on this interpretation have supported the proposal that the cytochrome is reduced at a rate sufficient to account for the rate of O2- production by activated neutrophils. We have confirmed the partial reduction of cytochrome b558 in neutrophil cytoplasts and in oxidase preparations exposed to NADPH, but have found that the reduction of the cytochrome bears no apparent relation to the activity of the oxidase, and can occur when NADPH is added to neutrophil membrane preparations that are unable to manufacture O2-. We therefore conclude that the NADPH-dependent reduction of cytochrome b558 seen in these preparations is unlikely to be a reflection of a catalysis-related steady state and that inferences drawn from such observations regarding the kinetic competence of the cytochrome may need to be reconsidered.
Foroozan
R
R
Scripps Research Institute, La Jolla, California 92037.
Ruedi
J M
JM
Babior
B M
BM
D000469:Alkaline Phosphatase / Q000097:blood; D002462:Cell Membrane / Q000378:metabolism; D002474:Cell-Free System; D003573:Cytochrome b Group / Q000097:blood* / Q000172:deficiency; D003594:Cytoplasmic Granules / Q000378:metabolism; D003600:Cytosol / Q000378:metabolism; D006801:Humans; D007700:Kinetics; D009240:N-Formylmethionine Leucyl-Phenylalanine / Q000494:pharmacology; D009247:NADH, NADPH Oxidoreductases / Q000097:blood*; D009249:NADP / Q000378:metabolism; D019255:NADPH Oxidases; D009504:Neutrophils / Q000201:enzymology*; D010084:Oxidation-Reduction; D010703:Phorbol Esters / Q000494:pharmacology; D013053:Spectrophotometry; D013481:Superoxides / Q000097:blood; D014155:Transcobalamins / Q000378:metabolism
D003573:Cytochrome b Group; D010703:Phorbol Esters; D014155:Transcobalamins; D013481:Superoxides; D009249:NADP; D009240:N-Formylmethionine Leucyl-Phenylalanine; C036888:cytochrome b558; D009247:NADH, NADPH Oxidoreductases; D019255:NADPH Oxidases; C022709:superoxide-forming enzyme; D000469:Alkaline Phosphatase