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Interaction of p67phox with Prdx6 regulates PLA2 activity
Author(s) -
Krishnaiah Saikumari Yegnisettipalli,
Dodia Chandra,
Chatterjee Shampa,
Feinstein Sheldon I,
Fisher Aron B
Publication year - 2011
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/fasebj.25.1_supplement.lb70
Subject(s) - nadph oxidase , superoxide , immunoprecipitation , polyclonal antibodies , chemistry , microbiology and biotechnology , phosphorylation , phospholipase a2 , angiotensin ii , in vitro , phospholipase , biochemistry , enzyme , biology , antibody , receptor , immunology , gene
Phospholipase A 2 (PLA 2 ) activity of peroxiredoxin 6 (Prdx6) is necessary to activate NADPH oxidase (NOX2). P67 phox is an essential component of the NOX2 enzyme complex that produces superoxide ions in response to agonist stimulation. It has been previously reported that p67 phox interacts with Prdx6 in PMN. We further investigated the interaction of these two proteins and its physiological role. In vitro assays used recombinant proteins and phosphoPrdx6 that was generated by incubation of Prdx6 with Erk2 plus ATP. His‐tagged Prdx6 pulled down p67 phox and his‐tagged p67 phox pulled‐down Prdx6 and phosphorylated Prdx6. A monoclonal anti‐p67 phox Ab immunoprecipitated Prdx6 and phosphoPrdx6, while a polyclonal anti‐Prdx6 Ab immunoprecipitated p67 phox . Phosphorylation of Prdx6 increased its apparent binding to p67 phox . Prdx6 and phosphoPrdx6 also were co‐immunoprecipitated from lysates of angiotensin II‐stimulated mouse pulmonary microvascular endothelial cells by anti‐ p67phox antibody. PLA 2 activity, measured at pH 7 in Ca 2+ ‐free buffer was 50 nmol/min/mg for Prdx6 and 1303 nmol/min/mg for phosphoPrdx6, but only 14–17 nmol/min/mg in the presence of p67 phox . Our data indicate that p67 phox binds to Prdx6 and inhibits its PLA 2 activity and thus could function to terminate the PLA 2 ‐mediated NOX2 activation signal.