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Mitigation of NADPH Oxidase 2 Activity as a Strategy to Inhibit Peroxynitrite Formation. J Biol Chem 2016 Mar 25;291(13):7029-44

Date

02/04/2016

Pubmed ID

26839313

Pubmed Central ID

PMC4807286

DOI

10.1074/jbc.M115.702787

Scopus ID

2-s2.0-84964907042 (requires institutional sign-in at Scopus site)   56 Citations

Abstract

Using high throughput screening-compatible assays for superoxide and hydrogen peroxide, we identified potential inhibitors of the NADPH oxidase (Nox2) isoform from a small library of bioactive compounds. By using multiple probes (hydroethidine, hydropropidine, Amplex Red, and coumarin boronate) with well defined redox chemistry that form highly diagnostic marker products upon reaction with superoxide (O2 (̇̄)), hydrogen peroxide (H2O2), and peroxynitrite (ONOO(-)), the number of false positives was greatly decreased. Selected hits for Nox2 were further screened for their ability to inhibit ONOO(-)formation in activated macrophages. A new diagnostic marker product for ONOO(-)is reported. We conclude that the newly developed high throughput screening/reactive oxygen species assays could also be used to identify potential inhibitors of ONOO(-)formed from Nox2-derived O2 (̇̄)and nitric oxide synthase-derived nitric oxide.

Author List

Zielonka J, Zielonka M, VerPlank L, Cheng G, Hardy M, Ouari O, Ayhan MM, Podsiadły R, Sikora A, Lambeth JD, Kalyanaraman B

Authors

Gang Cheng PhD Assistant Professor in the Biophysics department at Medical College of Wisconsin
Micael Joel Hardy PhD Visiting Assistant Professor in the Biophysics department at Medical College of Wisconsin
Balaraman Kalyanaraman PhD Professor in the Biophysics department at Medical College of Wisconsin
Jacek M. Zielonka PhD Assistant Professor in the Biophysics department at Medical College of Wisconsin




MESH terms used to index this publication - Major topics in bold

Chromatography, High Pressure Liquid
Coumarins
Enzyme Inhibitors
Fluorometry
Gene Expression
HL-60 Cells
High-Throughput Screening Assays
Humans
Hydrogen Peroxide
Macrophage Activation
Membrane Glycoproteins
Molecular Probes
NADPH Oxidase 2
NADPH Oxidases
Oxazines
Oxidation-Reduction
Peroxynitrous Acid
Phenanthridines
Quaternary Ammonium Compounds
Small Molecule Libraries
Superoxides
Tetradecanoylphorbol Acetate