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  • Title: Immunoaffinity purification of an oxidase-activating cytosolic complex from bovine neutrophils.
    Author: Jouan A, Dagher MC, Fuchs A, Foucaud-Gamen J, Vignais PV.
    Journal: Biochem Biophys Res Commun; 1993 Dec 30; 197(3):1296-302. PubMed ID: 8280146.
    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.
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