The membrane fraction and three cytosolic proteins of neutrophils, p47-phox, p67-phox and a G-protein, are involved in the cell-free activation of the O2--generating NADPH oxidase in the presence of SDS, though it has been controversial whether the G-protein is required or just enhancing the activity. We have used the three cytosolic factors, the solubilized membrane fraction, GTPγS and SDS, and found that both G-protein and GTPγS are essential for the activation of the NADPH oxidase. The effect of GTPγS is modified by Mg2+: the cations enhance the O2- generation at low concentrations of GTPγS, whereas they attenuate the activity at higher concentrations of GTPγS. In the presence of 10 μM GTPγS, the maximal activity is observed at 0.1 μM Mg2+, which is several-fold higher than that at 1 mM Mg2+. The omission of Mg2+ followed by the chelation with EDTA results in loss of the activation, which is completely restored by the addition of Mg2+. Thus, Mg2+ seems to modulate the activation of the NADPH oxidase at the level of the G-protein.
|Number of pages||7|
|Journal||Biochemical and Biophysical Research Communications|
|Publication status||Published - Jul 15 1992|
All Science Journal Classification (ASJC) codes
- Molecular Biology
- Cell Biology