Protein phosphorylation in neutrophils from patients with p67-phox-deficient chronic granulomatous disease

Neutrophils are known to contain a major 67-kD protein that undergoes enhanced phosphorylation and translocation to the membrane during cell stimulation, Recent studies have assumed that this 67-kD phosphoprotein is the 67-kD subunit of the phagocyte oxidase (p67-phox), We compare here the protein phosphorylation patterns in lysates of normal neutrophils and neutrophils from patients with chronic granulomatous disease (CGD) that are completely deficient in p67-phox, The phosphoproteins were labeled by incubation of the cells with radioactive inorganic phosphate (P-32(i)) or by the addition of [gamma-P-32]ATP to electropermeabilized neutrophils. With either method, stimulation of the normal or CGD cells always resulted in an enhanced incorporation of P-32 into two proteins in the 67-kD area, The extent of phosphorylation of these two proteins was very similar in the normal and CGD cells when permeabilized neutrophils loaded with [gamma-P-32]ATP were compared, Moreover, no overall differences in the protein phosphorylation patterns were observed between the normal and CGD cells, Our data indicate that the major 67-kD phosphoproteins observed in stimulated neutrophils are clearly different from p67-phox. (C) 1996 by The American Society of Hematology.