TY - JOUR
T1 - Nox2 B-loop peptide, Nox2ds, specifically inhibits the NADPH oxidase Nox2
AU - Csányi, Gábor
AU - Cifuentes-Pagano, Eugenia
AU - Al Ghouleh, Imad
AU - Ranayhossaini, Daniel J.
AU - Egaña, Loreto
AU - Lopes, Lucia R.
AU - Jackson, Heather M.
AU - Kelley, Eric E.
AU - Pagano, Patrick J.
N1 - Funding Information:
This work was supported by National Institutes of Health Grants HL079207 and HL55425. We thank Sheila Frizzell for her critical review of the manuscript. P.J.P. is an Established Investigator of the American Heart Association. P.J.P. receives research support from the Vascular Medicine Institute, the Institute for Transfusion Medicine, and the Hemophilia Center of Western Pennsylvania. G.C. is a recipient of an American Heart Association Postdoctoral Fellowship. The authors wish to sincerely thank Dr. Edgar Pick for his critical review of our data as well as methodological advice.
PY - 2011/9/15
Y1 - 2011/9/15
N2 - In recent years, reactive oxygen species (ROS) derived from the vascular isoforms of NADPH oxidase, Nox1, Nox2, and Nox4, have been implicated in many cardiovascular pathologies. As a result, the selective inhibition of these isoforms is an area of intense current investigation. In this study, we postulated that Nox2ds, a peptidic inhibitor that mimics a sequence in the cytosolic B-loop of Nox2, would inhibit ROS production by the Nox2-, but not the Nox1- and Nox4-oxidase systems. To test our hypothesis, the inhibitory activity of Nox2ds was assessed in cell-free assays using reconstituted systems expressing the Nox2-, canonical or hybrid Nox1-, or Nox4-oxidase. Our findings demonstrate that Nox2ds, but not its scrambled control, potently inhibited superoxide (O 2 •-) production in the Nox2 cell-free system, as assessed by the cytochrome c assay. Electron paramagnetic resonance confirmed that Nox2ds inhibits O 2 •- production by Nox2 oxidase. In contrast, Nox2ds did not inhibit ROS production by either Nox1- or Nox4-oxidase. These findings demonstrate that Nox2ds is a selective inhibitor of Nox2-oxidase and support its utility to elucidate the role of Nox2 in organ pathophysiology and its potential as a therapeutic agent.
AB - In recent years, reactive oxygen species (ROS) derived from the vascular isoforms of NADPH oxidase, Nox1, Nox2, and Nox4, have been implicated in many cardiovascular pathologies. As a result, the selective inhibition of these isoforms is an area of intense current investigation. In this study, we postulated that Nox2ds, a peptidic inhibitor that mimics a sequence in the cytosolic B-loop of Nox2, would inhibit ROS production by the Nox2-, but not the Nox1- and Nox4-oxidase systems. To test our hypothesis, the inhibitory activity of Nox2ds was assessed in cell-free assays using reconstituted systems expressing the Nox2-, canonical or hybrid Nox1-, or Nox4-oxidase. Our findings demonstrate that Nox2ds, but not its scrambled control, potently inhibited superoxide (O 2 •-) production in the Nox2 cell-free system, as assessed by the cytochrome c assay. Electron paramagnetic resonance confirmed that Nox2ds inhibits O 2 •- production by Nox2 oxidase. In contrast, Nox2ds did not inhibit ROS production by either Nox1- or Nox4-oxidase. These findings demonstrate that Nox2ds is a selective inhibitor of Nox2-oxidase and support its utility to elucidate the role of Nox2 in organ pathophysiology and its potential as a therapeutic agent.
KW - Cardiovascular disease
KW - Free radicals
KW - NADPH oxidase
KW - Nox inhibitor
KW - Reactive oxygen species
KW - Superoxide
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U2 - 10.1016/j.freeradbiomed.2011.04.025
DO - 10.1016/j.freeradbiomed.2011.04.025
M3 - Article
C2 - 21586323
AN - SCOPUS:80051802409
SN - 0891-5849
VL - 51
SP - 1116
EP - 1125
JO - Free Radical Biology and Medicine
JF - Free Radical Biology and Medicine
IS - 6
ER -