Superoxide production of reisolated membranes in the cell-free system
| . | Membranes From Patient With Control Cytosol . | Cytosol From Patient With Control Membranes . | Membranes and Cytosol From Control . |
|---|---|---|---|
| JW (T341K) | 0.06 (±0.06) | 4.17 (±0.37) | 4.11 (±0.18) |
| CE (C369R) | 0.21 (±0.01) | 3.13 (±0.18) | 2.89 (±0.34) |
| MK (G408E) | 0.02 (±0.03) | 2.77 (±0.21) | 2.99 (±0.61) |
| DG (E568K) | 0.08 (±0.05) | 3.35 (±0.25) | 3.39 (±0.32) |
| . | Membranes From Patient With Control Cytosol . | Cytosol From Patient With Control Membranes . | Membranes and Cytosol From Control . |
|---|---|---|---|
| JW (T341K) | 0.06 (±0.06) | 4.17 (±0.37) | 4.11 (±0.18) |
| CE (C369R) | 0.21 (±0.01) | 3.13 (±0.18) | 2.89 (±0.34) |
| MK (G408E) | 0.02 (±0.03) | 2.77 (±0.21) | 2.99 (±0.61) |
| DG (E568K) | 0.08 (±0.05) | 3.35 (±0.25) | 3.39 (±0.32) |
Values in μmol /min (mean ± SD; n = 3). Patients' or control neutrophil membranes (20 μg) were incubated with control cytosol (400 μg protein) in oxidase buffer at room temperature. After 10 minutes of incubation in the presence of SDS and GTPγS, membranes were reisolated from each incubation mixture by sucrose gradient centrifugation. NADPH oxidase activity was determined in one-fifth part of the membrane fractions by incubation in oxidase buffer in the presence of SDS (100 μmol/L), GTPγS (10 μmol/L), and NADPH (250 μmol/L) by the rate of cytochrome c reduction.