Abstract
Phospholipase A2 (PLA2) activity of peroxiredoxin 6 (Prdx6) is necessary to activate NADPH oxidase (NOX2). P67phox is an essential component of the NOX2 enzyme complex that produces superoxide ions in response to agonist stimulation. It has been previously reported that p67phox 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 p67phox and his‐tagged p67phox pulled‐down Prdx6 and phosphorylated Prdx6. A monoclonal anti‐p67phox Ab immunoprecipitated Prdx6 and phosphoPrdx6, while a polyclonal anti‐Prdx6 Ab immunoprecipitated p67phox. Phosphorylation of Prdx6 increased its apparent binding to p67phox. Prdx6 and phosphoPrdx6 also were co‐immunoprecipitated from lysates of angiotensin II‐stimulated mouse pulmonary microvascular endothelial cells by anti‐ p67phox antibody. PLA2 activity, measured at pH 7 in Ca2+‐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 p67phox. Our data indicate that p67phox binds to Prdx6 and inhibits its PLA2 activity and thus could function to terminate the PLA2 ‐mediated NOX2 activation signal.
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a callDisclaimer: All third-party content on this website/platform is and will remain the property of their respective owners and is provided on "as is" basis without any warranties, express or implied. Use of third-party content does not indicate any affiliation, sponsorship with or endorsement by them. Any references to third-party content is to identify the corresponding services and shall be considered fair use under The CopyrightLaw.
