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Inhibition of NADPH oxidase 5 activity by NO
Author(s) -
Qian Jin,
Fulton David
Publication year - 2010
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.24.1_supplement.785.16
Subject(s) - peroxynitrite , nadph oxidase , chemistry , enos , biochemistry , enzyme , ionomycin , nitric oxide , enzyme activator , enzyme assay , microbiology and biotechnology , superoxide , nitric oxide synthase , biology , in vitro , organic chemistry
The NADPH oxidases (Nox) are a family of multi‐subunit enzymes that produce superoxide and other reactive oxygen species (ROS). Nox5 is a calcium‐dependent enzyme that does not depend on accessory subunits for activation and in comparison to other Nox isoforms; the molecular regulation of Nox5 is poorly understood. Recently we and others have documented the expression of Nox5 in human blood vessels and the goal of the current study was to determine whether NO can modulate Nox5 activity. Endogenous NO from iNOS was a potent inhibitor of basal and both ionomycin and PMA‐stimulated activity. This action of iNOS was reversible with chronic, but not acute, exposure to L‐NAME and was due to the diffusion of NO, as iNOS was able to inhibit Nox5 when transfected in separate cells. Nox5 activity was also inhibited by eNOS and PM‐targeted eNOS albeit to a lesser extent. The ability of NO to inhibit Nox5 activity was due to a direct modification of enzyme activity as Nox5 activity was inhibited in a disrupted/cell free enzyme activity assay replete with excess calcium and NADPH. Co‐expression of iNOS and Nox5 did not increase the levels of ONOO‐ or induce the tyrosine nitration of Nox5, excluding a role for peroxynitrite. In contrast, there was evidence for the increased nitrosylation of Nox5 using the biotin‐switch assay. Collectively, these results suggest that endogenous NO can directly nitrosylate and inhibit the activity of Nox5.