UCL Discovery

Abstract

The NADPH oxidase generates microbicidal superoxide in phagocytes, and when defective it leads to chronic granulomatous disease (CGD), Oxidase specific proteins in the cytosol, p47(phox) and p67(phox), 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 he regulated by cytoskeletal proteins, we examined the cytosolic phox proteins for interactions with cytoskeletal elements, p67(phox) 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 p40(phox) a binding partner of p67(phox). 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 p47(phox) or p67(phox) 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.