Conversion of NOX2 into a constitutive enzyme in vitro and in living cells, after its binding with a chimera of the regulatory subunits.

Conversion of NOX2 into a constitutive enzyme in vitro and in living cells, after its binding with a chimera of the regulatory subunits.
Free Radic Biol Med. 2017 Oct 24;:
Authors: Masoud R, Serfaty X, Erard M, Machillot P, Karimi G, Hudik E, Wien F, Baciou L, Levin CH, Bizouarn T
Abstract
During the phagocytosis of pathogens by phagocyte cells, the NADPH oxidase complex is activated to produce superoxide anion, a precursor of microbial oxidants. The activated NADPH oxidase complex from phagocytes consists in two transmembrane proteins (Nox2 and p22(phox)) and four cytosolic proteins (p40(phox), p47(phox), p67(phox) and Rac1-2). In the resting state of the cells, these proteins are dispersed in the cytosol, the membrane of granules and the plasma membrane. In order to synchronize the assembly of the cytosolic subunits on the membrane components of the oxidase, a fusion of the cytosolic proteins p47(phox), p67(phox) and Rac1 named trimera was constructed. The trimera investigated in this paper is composed of the p47(phox) segment 1-286, the p67(phox) segment 1-212 and the mutated Rac1(Q61L). We demonstrate that the complex trimera-cyt b558 is functionally comparable to the one containing the separated subunits. Each of the subunits p47(phox), p67(phox) and Rac1Q61L has kept its own activating property. The trimera is produced in an activated conformation as seen by circular dichroism. However, the presence of amphiphile is still neces...

Source: Free Radical Biology and Medicine - October 24, 2017 Category: Biology Authors: Masoud R, Serfaty X, Erard M, Machillot P, Karimi G, Hudik E, Wien F, Baciou L, Levin CH, Bizouarn T Tags: Free Radic Biol Med Source Type: research