Nitrosopersulfide (SSNO(-)) accounts for sustained NO bioactivity of S-nitrosothiols following reaction with sulfide

Redox Biol. 2014 Jan 11;2:234-44. doi: 10.1016/j.redox.2013.12.031. eCollection 2014.


Sulfide salts are known to promote the release of nitric oxide (NO) from S-nitrosothiols and potentiate their vasorelaxant activity, but much of the cross-talk between hydrogen sulfide and NO is believed to occur via functional interactions of cell regulatory elements such as phosphodiesterases. Using RFL-6 cells as an NO reporter system we sought to investigate whether sulfide can also modulate nitrosothiol-mediated soluble guanylyl cyclase (sGC) activation following direct chemical interaction. We find a U-shaped dose response relationship where low sulfide concentrations attenuate sGC stimulation by S-nitrosopenicillamine (SNAP) and cyclic GMP levels are restored at equimolar ratios. Similar results are observed when intracellular sulfide levels are raised by pre-incubation with the sulfide donor, GYY4137. The outcome of direct sulfide/nitrosothiol interactions also critically depends on molar reactant ratios and is accompanied by oxygen consumption. With sulfide in excess, a 'yellow compound' accumulates that is indistinguishable from the product of solid-phase transnitrosation of either hydrosulfide or hydrodisulfide and assigned to be nitrosopersulfide (perthionitrite, SSNO(-); λ max 412 nm in aqueous buffers, pH 7.4; 448 nm in DMF). Time-resolved chemiluminescence and UV-visible spectroscopy analyses suggest that its generation is preceded by formation of the short-lived NO-donor, thionitrite (SNO(-)). In contrast to the latter, SSNO(-) is rather stable at physiological pH and generates both NO and polysulfides on decomposition, resulting in sustained potentiation of SNAP-induced sGC stimulation. Thus, sulfide reacts with nitrosothiols to form multiple bioactive products; SSNO(-) rather than SNO(-) may account for some of the longer-lived effects of nitrosothiols and contribute to sulfide and NO signaling.

Keywords: CysNO, S-nitrosocysteine; DMF, dimetylformamide; DMSO, dimethylsulfoxide; GSNO, S-nitrosoglutathione; HSNO; Hydrogen sulfide; IPN, isopentyl nitrite; NO+, nitrosonium; NO, nitric oxide; Nitric oxide; Nitroxyl; Polysulfides; RFL-6, rat fibroblastoid-like cell line; SNAP, S-nitrosopenicillamine; SNO−, thionitrite; SSNO−, nitrosopersulfide, perthionitrite, PDE, phopsphodiesterase; cGMP; sGC, soluble guanylyl cyclase.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Cell Line
  • Cyclic GMP / metabolism
  • Guanylate Cyclase / metabolism*
  • Morpholines / pharmacology
  • Nitric Oxide / metabolism*
  • Nitric Oxide Donors / pharmacology
  • Nitroso Compounds / pharmacology
  • Organothiophosphorus Compounds / pharmacology
  • Oxygen Consumption / drug effects
  • Rats
  • Receptors, Cytoplasmic and Nuclear / metabolism*
  • S-Nitrosothiols / metabolism*
  • Soluble Guanylyl Cyclase
  • Sulfides / metabolism*


  • GYY 4137
  • Morpholines
  • Nitric Oxide Donors
  • Nitroso Compounds
  • Organothiophosphorus Compounds
  • Receptors, Cytoplasmic and Nuclear
  • S-Nitrosothiols
  • Sulfides
  • persulfides
  • Nitric Oxide
  • S-nitrosopenicillamine
  • Guanylate Cyclase
  • Soluble Guanylyl Cyclase
  • Cyclic GMP