Targeting NADPH-oxidase by reactive oxygen species reveals an initial sensitive step in the assembly process.
NADPH-oxidase (Nox) is a highly regulated dynamic complex comprising membrane and cytosolic proteins and is the major source of nonmitochondrial cellular reactive oxygen species (ROS). In phagocyte cells, in which ROS are produced in huge amounts, Nox is "naturally" assailed by the action of its own ROS. We have subjected each individual component of Nox or the whole complex at various times during the assembly process either to oxygen free radicals produced by radiolysis or to hydrogen peroxide. Membrane components presented the highest irradiation sensitivity. Irradiation of p67(phox) drastically decreased its interaction with arachidonic acid and destabilized the [p47(phox)-p67(phox)] complex. When the system was irradiated during its assembly process, we could identify an initial irradiation-sensitive phase followed by a resistant form when the complex was assembled.
Ostuni
Mariano A
MA
Laboratoire de Chimie Physique, UMR 8000, Université Paris-Sud, F-91405 Orsay, France. mariano.ostuni@upmc.fr
Gelinotte
Magali
M
Bizouarn
Tania
T
Baciou
Laura
L
Houée-Levin
Chantal
C
D000818:Animals; D002417:Cattle; D004351:Drug Resistance / Q000187:drug effects / Q000502:physiology; D004789:Enzyme Activation / Q000187:drug effects / Q000528:radiation effects; D006801:Humans; D006861:Hydrogen Peroxide / Q000494:pharmacology; D008956:Models, Chemical; D019255:NADPH Oxidases / Q000037:antagonists & inhibitors / Q000378:metabolism* / Q000528:radiation effects; D010750:Phosphoproteins / Q000378:metabolism; D055503:Protein Multimerization / Q000187:drug effects*; D021122:Protein Subunits / Q000187:drug effects / Q000378:metabolism / Q000528:radiation effects; D017382:Reactive Oxygen Species / Q000494:pharmacology*
D010750:Phosphoproteins; D021122:Protein Subunits; D017382:Reactive Oxygen Species; C079789:neutrophil cytosol factor 67K; D006861:Hydrogen Peroxide; D019255:NADPH Oxidases; C060505:neutrophil cytosolic factor 1