Purification and properties of a functional 47-kilodalton cytosolic factor required for NADPH-oxidase activation in bovine neutrophils.
A cytosolic factor of 47 kDa required for activation of the NADPH oxidase, and referred to as p47, has been purified in its functional form from the cytosol of resting bovine neutrophils. The purification was monitored by the determination of the activating potency of p47 in a cell-free system of oxidase activation. The recovery was around 10% and the purification factor greater than 1000. P47 was phosphorylated in vitro by protein kinase A and protein kinase C. [32P] labeled p47 was resolved by isoelectric focusing into two major labeled bands of pI 7.0 and 8.5. Polyclonal antibodies were used to demonstrate that p47 is localized specifically in the cytosol of resting neutrophils, and that, upon activation of neutrophils, p47 is translocated from the cytosol to the membrane.
Pilloud-Dagher
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MC
Départment de Biologie Moléculaire et Structurale, Centre d'Etudes Nucléaires, Grenoble, France.
Jouan
A
A
Vignais
P V
PV
D000818:Animals; D001798:Blood Proteins / Q000302:isolation & purification / Q000502:physiology*; D002417:Cattle; D002474:Cell-Free System; D002848:Chromatography, DEAE-Cellulose; D002852:Chromatography, Ion Exchange; D003600:Cytosol / Q000378:metabolism; D004789:Enzyme Activation; D007700:Kinetics; D008970:Molecular Weight; D009247:NADH, NADPH Oxidoreductases / Q000097:blood*; D019255:NADPH Oxidases; D009504:Neutrophils / Q000201:enzymology*; D010766:Phosphorylation; D011493:Protein Kinase C / Q000378:metabolism; D011494:Protein Kinases / Q000378:metabolism
D001798:Blood Proteins; D009247:NADH, NADPH Oxidoreductases; D019255:NADPH Oxidases; D011494:Protein Kinases; D011493:Protein Kinase C