title: Immunoaffinity purification of an oxidase-activating cytosolic complex from bovine neutrophils.
abstract: An oxidase activating complex from the cytosol of bovine neutrophils was purified by immunoaffinity using a monoclonal antibody specific for the 67 kDa cytosolic factor of oxidase activation (p67) and assayed for production of superoxide O2- in a cell-free system. The complex comprised not only p67, but also the second cytosolic factor of 47 kDa (p47) in equivalent amounts. The p47-p67 complex showed a good oxidase activating potency when added to neutrophil membranes in the presence of GTP-gamma-S and arachidonic acid. A ras-related small G protein could not be immunodetected in the p47-p67 activating complex, indicating that the GTP required for oxidase activation in the cell free system bound to a protein that was either present in catalytic amounts in the cytosolic complex or present in sufficient amount in the membrane fraction.
authors.affiliation: Laboratoire de Biochimie (URA 1130/CNRS), Département de Biologie Moléculaire et Structurale Grenoble, France.
authors.forename: A
authors.initials: A
authors.lastname: Jouan
authors.forename: M C
authors.initials: MC
authors.lastname: Dagher
authors.forename: A
authors.initials: A
authors.lastname: Fuchs
authors.forename: J
authors.initials: J
authors.lastname: Foucaud-Gamen
authors.forename: P V
authors.initials: PV
authors.lastname: Vignais
mesh_terms: D000818:Animals; D015153:Blotting, Western; D002417:Cattle; D002474:Cell-Free System; D002846:Chromatography, Affinity; D003600:Cytosol / Q000201:enzymology; D004591:Electrophoresis, Polyacrylamide Gel; D008970:Molecular Weight; D009247:NADH, NADPH Oxidoreductases / Q000097:blood* / Q000302:isolation & purification; D019255:NADPH Oxidases; D009504:Neutrophils / Q000201:enzymology*; D013481:Superoxides / Q000097:blood
chemical_list: D013481:Superoxides; D009247:NADH, NADPH Oxidoreductases; D019255:NADPH Oxidases