A novel cytosolic component, p40phox, of respiratory burst oxidase associates with p67phox and is absent in patients with chronic granulomatous disease who lack p67phox.
Cytosolic components of human neutrophils, p47phox and p67phox, deficiencies of which lead to chronic granulomatous disease (CGD), potentiate respiratory burst oxidase translocating from cytosol to membrane upon cell stimulation. In this report we describe a novel cytosolic component, p40phox, which consistently behaves with p67phox through immunoprecipitation and column works, and is missing in patients with CGD who lack p67phox. Although actin has been reported to be involved in O2- generation, the p40phox profile did not correspond to that of actin. The tight association between p40phox and p67phox was not affected by treatment with a mixture of deoxycholate and Nonidet P-40, until subjected to SDS-PAGE. Addition of recombinant p67phox to cytosol did not produce any additional p40phox in the immunoprecipitate, unlike the additive increment in the band of p67phox. These results suggest that p40phox forms a complex with p67phox in a molar ratio of 1:1, without any free p40phox in the cytosol.
Tsunawaki
S
S
National Children's Medical Research Center, Tokyo, Japan.
Mizunari
H
H
Nagata
M
M
Tatsuzawa
O
O
Kuratsuji
T
T
D002462:Cell Membrane / Q000201:enzymology; D002852:Chromatography, Ion Exchange; D003600:Cytosol / Q000201:enzymology; D004591:Electrophoresis, Polyacrylamide Gel; D006105:Granulomatous Disease, Chronic / Q000097:blood / Q000201:enzymology*; D006801:Humans; D015151:Immunoblotting; D009247:NADH, NADPH Oxidoreductases / Q000097:blood*; D019255:NADPH Oxidases; D009504:Neutrophils / Q000201:enzymology*; D010750:Phosphoproteins / Q000097:blood* / Q000172:deficiency* / Q000302:isolation & purification; D012016:Reference Values
D010750:Phosphoproteins; C085072:neutrophil cytosol factor 40K; C079789:neutrophil cytosol factor 67K; D009247:NADH, NADPH Oxidoreductases; D019255:NADPH Oxidases; C022709:superoxide-forming enzyme