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p67 phox terminates the phospholipase A 2 ‐derived signal for activation of NADPH oxidase (NOX2)
Author(s) -
Krishnaiah Saikumari Y.,
Dodia Chandra,
Feinstein Sheldon I.,
Fisher Aron B.
Publication year - 2013
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fj.12-222133
Subject(s) - nadph oxidase , chemistry , phospholipase a2 , phospholipase , angiotensin ii , microbiology and biotechnology , oxidase test , enzyme , biochemistry , biology , receptor
The phospholipase A 2 (PLA 2 )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. PLA 2 activity of recombinant phosphoPrdx6 was decreased by >98% in the presence of p67 phox ; the calculated dissociation constant ( K d ) of the p67 phox : phosphoPrdx6 complex was 65 nM. PLA 2 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 PLA 2 activity, an interaction that could function to terminate the PLA 2 ‐mediated NOX2 activation signal.—Krishnaiah, S. Y., Dodia, C., Feinstein, S. I., and Fisher, A. B. p67 phox terminates the phospholipase A 2 ‐derived signal for activation of NADPH oxidase (NOX2). FASEB J. 27, 2066–2073 (2013). www.fasebj.org