Purification of the 28.5 kDa cytosolic protein involved in the activation of NADPH oxidase from guinea pig neutrophils.
We tried to purify a new protein component required for the activation of NADPH oxidase from the guinea pig neutrophil cytosolic fraction which did not contain p47phox and p67phox, using HAC-5CP, IEC-QA and Superose 12HR columns. The NADPH oxidase-activating activity was separated into three fractions on IEC-QA anion-exchange HPLC. However, when each of the fractions was purified by Superose 12HR gel filtration, the active fraction eluted at the same position, and was found to contain a common protein with a molecular weight of 28.5 kDa on SDS-PAGE. These results suggest that the 28.5 kDa protein is a novel NADPH oxidase activating protein.
Someya
A
A
Department of Biochemistry, Juntendo University, School of Medicine, Tokyo, Japan.
Yomogida
S
S
Nagaoka
I
I
Iwabuchi
K
K
Yamashita
T
T
D000818:Animals; D002851:Chromatography, High Pressure Liquid; D003600:Cytosol / Q000378:metabolism*; D004591:Electrophoresis, Polyacrylamide Gel; D004789:Enzyme Activation; D006168:Guinea Pigs; D015151:Immunoblotting; D009247:NADH, NADPH Oxidoreductases / Q000302:isolation & purification / Q000378:metabolism*; D019255:NADPH Oxidases; D009504:Neutrophils / Q000201:enzymology*
D009247:NADH, NADPH Oxidoreductases; D019255:NADPH Oxidases