Recovery of reduced thiol groups by superoxide-mediated denitrosation of nitrosothiols

Stefan Schildknecht, Alex Von Kriegsheim, Ksenija Vujacic-mirski, Fabio Di Lisa, Volker Ullrich, Andreas Daiber

Research output: Contribution to journalArticlepeer-review

Abstract

Nitrosation of critical thiols has been elaborated as reversible posttranslational modification with regulatory function in multiple disorders. Reversibility of S-nitrosation is generally associated with enzyme-mediated oneelectron reductions, catalyzed by the thioredoxin system, or by nitrosoglutathione reductase. In the present study, we confirm previous evidence for a non-enzymatic de-nitrosation of nitrosoglutathione (GSNO) by superoxide. The interaction leads to the release of nitric oxide that subsequently interacts with a second molecule of superoxide (O2•􀀀 ) to form peroxynitrite. Despite the formation of peroxynitrite, approximately 40–70% of GSNO yielded reduced glutathione (GSH), depending on the applied analytical assay. The concept of O2•􀀀 dependent denitrosation was then applied to S-nitrosated enzymes. S-nitrosation of isocitrate dehydrogenase (ICDH; NADP+-dependent) was accompanied by an inhibition of the enzyme and could be reversed by dithiothreitol. Treatment of nitrosated ICDH with O2•􀀀 indicated ca. 50% recovery of enzyme activity. Remaining inhibition was largely consequence of oxidative modifications evoked either by O2•􀀀 or by peroxynitrite. Recovery of activity in S-nitrosated enzymes by O2•􀀀 appears relevant only for selected examples. In contrast, recovery of reduced glutathione from the interaction of GSNO with O2•􀀀 could represent a mechanism to regain reducing equivalents in situations of excess O2•􀀀 formation, e.g. in the reperfusion phase after ischemia.
Original languageEnglish
Pages (from-to)102439
JournalRedox biology
Volume56
Early online date14 Aug 2022
DOIs
Publication statusE-pub ahead of print - 14 Aug 2022

Keywords

  • S-nitros(yl)ation
  • S-denitrosation
  • Superoxide
  • Nitric oxide
  • Ischemia/reperfusion

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