Redox modulation of endothelial nitric oxide synthase by glutaredoxin-1 through reversible oxidative post-translational modification

Chun An Chen, Francesco De Pascali, Ariel Basye, Craig Hemann, Jay L. Zweier

Research output: Contribution to journalArticle

Abstract

S-Glutathionylation is a redox-regulated modification that uncouples endothelial nitric oxide synthase (eNOS), switching its function from nitric oxide (NO) synthesis to O2- generation, and serves to regulate vascular function. While in vitro or in vivo eNOS S-glutathionylation with modification of Cys689 and Cys908 of its reductase domain is triggered by high levels of glutathione disulfide (GSSG) or oxidative thiyl radical formation, it remains unclear how this process may be reversed. Glutaredoxin-1 (Grx1), a cytosolic and glutathione-dependent enzyme, can reverse protein S-glutathionylation; however, its role in regulating eNOS S-glutathionylation remains unknown. We demonstrate that Grx1 in the presence of glutathione (GSH) (1 mM) reverses GSSG-mediated eNOS S-glutathionylation with restoration of NO synthase activity. Because Grx1 also catalyzes protein S-glutathionylation with an increased [GSSG]/[GSH] ratio, we measured its effect on eNOS S-glutathionylation when the [GSSG]/[GSH] ratio was >0.2, which can occur in cells and tissues under oxidative stress, and observed an increased level of eNOS S-glutathionylation with a marked decrease in eNOS activity without uncoupling. This eNOS S-glutathionylation was reversed with a decrease in the [GSSG]/[GSH] ratio to

Original languageEnglish (US)
Pages (from-to)6712-6723
Number of pages12
JournalBiochemistry®
Volume52
Issue number38
DOIs
StatePublished - Sep 24 2013
Externally publishedYes

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Glutaredoxins
Nitric Oxide Synthase Type III
Post Translational Protein Processing
Oxidation-Reduction
Glutathione Disulfide
Modulation
Protein S
Glutathione
Switching functions
Oxidative stress
Nitric Oxide Synthase
Restoration
Blood Vessels
Oxidoreductases
Nitric Oxide
Oxidative Stress
Tissue

ASJC Scopus subject areas

  • Biochemistry

Cite this

Redox modulation of endothelial nitric oxide synthase by glutaredoxin-1 through reversible oxidative post-translational modification. / Chen, Chun An; De Pascali, Francesco; Basye, Ariel; Hemann, Craig; Zweier, Jay L.

In: Biochemistry®, Vol. 52, No. 38, 24.09.2013, p. 6712-6723.

Research output: Contribution to journalArticle

Chen, Chun An ; De Pascali, Francesco ; Basye, Ariel ; Hemann, Craig ; Zweier, Jay L. / Redox modulation of endothelial nitric oxide synthase by glutaredoxin-1 through reversible oxidative post-translational modification. In: Biochemistry®. 2013 ; Vol. 52, No. 38. pp. 6712-6723.
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