Immunoaffinity Purification of an Oxidase-Activating Cytosolic Complex from Bovine Neutrophils
- Resource Type
- Authors
- Alain H. Fuchs; Alain Jouan; Pierre V. Vignais; J. Foucaudgamen; Marie-Claire Dagher
- Source
- Biochemical and Biophysical Research Communications
Biochemical and Biophysical Research Communications, Elsevier, 1993, 197 (3), pp.1296-302
Biochemical and Biophysical Research Communications, 1993, 197 (3), pp.1296-302
- Subject
- GTP'
Neutrophils
Blotting, Western
Biophysics
Small G Protein
MESH: Neutrophils
MESH: Superoxides
Biology
[SDV.IMM.II]Life Sciences [q-bio]/Immunology/Innate immunity
Biochemistry
Chromatography, Affinity
Cell-free system
03 medical and health sciences
chemistry.chemical_compound
Cytosol
MESH: Cytosol
Superoxides
MESH: Cell-Free System
Animals
MESH: Blotting, Western
NADH, NADPH Oxidoreductases
MESH: Animals
MESH: NADPH Oxidase
[SDV.IMM.II] Life Sciences [q-bio]/Immunology/Innate immunity
Molecular Biology
030304 developmental biology
chemistry.chemical_classification
0303 health sciences
Oxidase test
MESH: NADH, NADPH Oxidoreductases
Cell-Free System
Superoxide
MESH: Molecular Weight
030302 biochemistry & molecular biology
NADPH Oxidases
Biological activity
Cell Biology
MESH: Chromatography, Affinity
Molecular biology
Molecular Weight
MESH: Cattle
Enzyme
chemistry
Cattle
Electrophoresis, Polyacrylamide Gel
MESH: Electrophoresis, Polyacrylamide Gel
- Language
- ISSN
- 0006-291X
1090-2104
International audience; 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.