MECHANISMS OF NADPH OXIDASE ACTIVATION - TRANSLOCATION OF P40(PHOX), RAC1 AND RAC2 FROM THE CYTOSOL TO THE MEMBRANES IN HUMAN NEUTROPHILS LACKING P47(PHOX) OR P67(PHOX)

On neutrophil stimulation, the cytosolic components of NADPH oxidase, p67(phox), p47(phox), p40(phox), as well as the Ras-related G-proteins Rac1 and Rac2, are translocated from the cytosol to cell membranes wh ere they associate with a flavocytochrome b, forming a functional comp lex responsible for the production of oxygen radicals in phagocytes. I n this paper we show that (a) in neutrophils from a patient with a for m of chronic granulomatous disease (CGD) in which p67(phox) is absent, p47(phox) and Rac2, but not p40(phox) and Rac1 were translocated from the cytosol to the membrane on stimulation with formylmethionyl-leucy lphenylalanine (fMLP) or phorbol 12-myristate 13-acetate (PMA), (b) in neutrophils from a patient with a form of CGD in which p47(phox) is a bsent, p67(phox), p40(phox) and Rac1 failed to associate with the memb rane on stimulation with fMLP or PMA, whereas Rac2 was translocated as in normal neutrophils. We also show that in neutrophils from a patien t lacking p67(phox), the amount of cytosolic p40(phox) was decreased b y about 40 %. These findings indicate that, on neutrophil stimulation, p67(phox) mediates the translocation of p40(phox) and Rac1 from the c ytosol to cell membranes and that Rac2 associates with the membranes i ndependently of p47(phox) and p67(phox).