title: p67(phox) terminates the phospholipase A(2)-derived signal for activation of NADPH oxidase (NOX2).
abstract: The phospholipase A2 (PLA2)activity of phosphorylated peroxiredoxin 6 (Prdx6) is required for activation of NADPH oxidase (NOX2). We investigated the interaction of Prdx6 with p67(phox) and its effect on NOX2 activity. With the use of specific antibodies, coimmunoprecipitation of p67(phox) and phosphorylated Prdx6 was demonstrated with lysates of mouse pulmonary microvascular endothelial cells (MPMVECs) that were stimulated with angiotensin II; the interaction of p67(phox) with nonphosphorylated Prdx6 was relatively weak. Association of p67(phox) and phosphoPrdx6 in intact MPMVECs after angiotensin II stimulation was demonstrated by proximity ligation assay and was abolished by U0126, a MAP kinase inhibitor. By isothermal titration calorimetry, p67(phox) bound strongly to phosphoPrdx6 but bound poorly to Prdx6; phosphorylated p67(phox) did not bind to either Prdx6 or phosphoPrdx6. PLA2 activity of recombinant phosphoPrdx6 was decreased by >98% in the presence of p67(phox); the calculated dissociation constant (Kd) of the p67(phox): phosphoPrdx6 complex was 65 nM. PLA2 activity (MJ33 sensitive) in cell lysates following angiotensin II treatment of MPMVECs was increased by 85% following knockdown of p67(phox) with siRNA. These data indicate that p67(phox) binds to phosphoPrdx6 and inhibits its PLA2 activity, an interaction that could function to terminate the PLA2-mediated NOX2 activation signal.-Krishnaiah, S. Y., Dodia, C., Feinstein, S. I., and Fisher, A. B. p67(phox) terminates the phospholipase A2-derived signal for activation of NADPH oxidase (NOX2).
authors: Institute for Environmental Medicine, University of Pennsylvania Perelman School of Medicine, Philadelphia, PA 19104, USA.
authors: Saikumari Y
authors: SY
authors: Krishnaiah
authors: Chandra
authors: C
authors: Dodia
authors: Sheldon I
authors: SI
authors: Feinstein
authors: Aron B
authors: AB
authors: Fisher
mesh_terms: D000804:Angiotensin II; D000818:Animals; D002070:Butadienes / Q000494:pharmacology; D042783:Endothelial Cells / Q000502:physiology; D006801:Humans; D007700:Kinetics; D008562:Membrane Glycoproteins / Q000378:metabolism*; D051379:Mice; D000074662:NADPH Oxidase 2; D019255:NADPH Oxidases / Q000378:metabolism*; D009570:Nitriles / Q000494:pharmacology; D054465:Peroxiredoxin VI / Q000378:metabolism*; D064801:Phospholipase A2 Inhibitors; D010750:Phosphoproteins / Q000378:metabolism*; D010766:Phosphorylation
chemical_list: D002070:Butadienes; D008562:Membrane Glycoproteins; D009570:Nitriles; D064801:Phospholipase A2 Inhibitors; D010750:Phosphoproteins; C113580:U 0126; C079789:neutrophil cytosol factor 67K; D000804:Angiotensin II; D054465:Peroxiredoxin VI; C488842:Cybb protein, mouse; D000074662:NADPH Oxidase 2; D019255:NADPH Oxidases
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