Cytosolic phox proteins interact with and regulate the assembly of coronin in neutrophils.
The NADPH oxidase generates microbicidal superoxide in phagocytes, and when defective it leads to chronic granulomatous disease (CGD). Oxidase specific proteins in the cytosol, p47phox and p67phox, as well as the small GTP binding protein p21rac are important for activation of superoxide production. Because the activity of this oxidase is normally tightly restricted to the phagocytic vacuole, and its temporal and spatial organisation might be regulated by cytoskeletal proteins, we examined the cytosolic phox proteins for interactions with cytoskeletal elements. p67phox copurified with a 57 kDa protein, identified as coronin, an actin binding protein that is important for movement and phagocytosis in Dictyostelium. Binding studies revealed that coronin attaches to the C-terminal half of p40phox, a binding partner of p67phox. The phox proteins and coronin had a similar distribution in the cell, and both accumulated around the phagocytic vacuole. PMA activation of adherent neutrophils resulted in a major rearrangement of these proteins, and of actin, which were lost from the periphery of the cell and condensed around the nucleus. The rearrangement of F-actin and coronin in adherent cells, were absent, or markedly diminished, in cells from patients lacking p47phox or p67phox in which an abnormally large proportion of the coronin was present as part of a large complex. The cytosolic phox proteins might play a regulatory role in the reorganisation of the cytoskeleton accompanying superoxide generation.
Grogan
A
A
Department of Medicine, University College London, London WC1E 6JJ, UK.
Reeves
E
E
Keep
N
N
Wientjes
F
F
Totty
N F
NF
Burlingame
A L
AL
Hsuan
J J
JJ
Segal
A W
AW
D000199:Actins / Q000378:metabolism; D003600:Cytosol / Q000378:metabolism*; D004789:Enzyme Activation; D006801:Humans; D008840:Microfilament Proteins / Q000378:metabolism*; D019255:NADPH Oxidases / Q000378:metabolism; D009504:Neutrophils / Q000276:immunology / Q000378:metabolism*; D010587:Phagocytosis; D010750:Phosphoproteins / Q000378:metabolism*; D011485:Protein Binding; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology
D000199:Actins; D008840:Microfilament Proteins; D010750:Phosphoproteins; C079789:neutrophil cytosol factor 67K; C072061:coronin proteins; D019255:NADPH Oxidases; C060505:neutrophil cytosolic factor 1; D013755:Tetradecanoylphorbol Acetate