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 (32Pi) or by the addition of [gamma-32P]ATP to electropermeabilized neutrophils. With either method, stimulation of the normal or CGD cells always resulted in an enhanced incorporation of 32p 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 -32P]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.
Heyworth
P G
PG
Department of Molecular and Experimental Medicine, Scripps Research Institute, La Jolla, CA, USA.
Ding
J
J
Erickson
R W
RW
Lu
D J
DJ
Curnutte
J T
JT
Badwey
J A
JA
D000595:Amino Acid Sequence; D015180:Electrophoresis, Gel, Two-Dimensional; D006105:Granulomatous Disease, Chronic / Q000378:metabolism* / Q000473:pathology; D006801:Humans; D008565:Membrane Proteins / Q000302:isolation & purification / Q000378:metabolism*; D008969:Molecular Sequence Data; D008970:Molecular Weight; D009504:Neutrophils / Q000378:metabolism* / Q000473:pathology; D010750:Phosphoproteins / Q000172:deficiency* / Q000302:isolation & purification / Q000378:metabolism*; D010766:Phosphorylation; D011499:Protein Processing, Post-Translational
D008565:Membrane Proteins; D010750:Phosphoproteins; C079789:neutrophil cytosol factor 67K