Analysis of glutathione mediated S-(de)nitrosylation in complex biological matrices by immuno-spin trapping and identification of two novel substrates

Esha Sircar, Detcho A. Stoyanovsky, Timothy R. Billiar, Arne Holmgren, Rajib Sengupta*

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

3 Scopus citations

Abstract

The intracellular concentration of reduced glutathione (GSH) lies in the range of 1–10 mM, thereby indisputably making it the most abundant intracellular thiol. Such a copious amount of GSH makes it the most potent and robust cellular antioxidant that plays a crucial role in cellular defence against redox stress. The role of GSH as a denitrosylating agent is well established; in this study, we demonstrate GSH mediated denitrosylation of HepG2 cell-derived protein nitrosothiols (PSNOs), by a unique spin-trapping mechanism, using 5,5-dimethyl-1-pyrroline N-oxide (DMPO) as the spin trapping agent, followed by a western blot analysis. We also report our findings of two, hitherto unidentified substrates of GSH mediated S-denitrosylation, namely S-nitrosoglutaredoxin 1 (Grx1-SNO) and S-nitrosylated R1 subunit of ribonucleotide reductase (R1-SNO).

Original languageEnglish
Pages (from-to)26-30
Number of pages5
JournalNitric Oxide - Biology and Chemistry
Volume118
DOIs
StatePublished - 1 Jan 2022
Externally publishedYes

Keywords

  • Denitrosylation
  • Glutaredoxin
  • Glutathione
  • Nitric oxide
  • S-nitrosylation
  • Thioredoxin

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