Nitrosopersulfide (SSNO(-)) accounts for sustained NO bioactivity of S-nitrosothiols following reaction with sulfide.
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.
Cardiovascular Research Laboratory, Department of Cardiology, Pneumology and Angiology, Medical Faculty, Heinrich Heine University of Düsseldorf, Düsseldorf, Germany.
Miriam M
MM
Cortese-Krott
Clinical and Experimental Sciences, Faculty of Medicine, University of Southampton, Southampton General Hospital, Tremona Road, Southampton, UK.
Bernadette O
BO
Fernandez
Clinical and Experimental Sciences, Faculty of Medicine, University of Southampton, Southampton General Hospital, Tremona Road, Southampton, UK.
José L T
JL
Santos
Institute for Pharmacology and Toxicology, Ruhr-University Bochum, Bochum, Germany.
Evanthia
E
Mergia
Institute of Molecular Physiology and Genetics, Slovak Academy of Sciences, Bratislava, Slovak Republic.
Marian
M
Grman
Department of Molecular Immunology and Toxicology, National Institute of Oncology, Ráth György utca 7-9, Budapest, Hungary.
Péter
P
Nagy
Cardiovascular Research Laboratory, Department of Cardiology, Pneumology and Angiology, Medical Faculty, Heinrich Heine University of Düsseldorf, Düsseldorf, Germany.
Malte
M
Kelm
Medical School, University of St-Andrews, St-Andrews, Fife, Scotland.
Anthony
A
Butler
Clinical and Experimental Sciences, Faculty of Medicine, University of Southampton, Southampton General Hospital, Tremona Road, Southampton, UK.
Martin
M
Feelisch
D000818:Animals; D002460:Cell Line; D006152:Cyclic GMP / Q000378:metabolism; D006162:Guanylate Cyclase / Q000378:metabolism*; D009025:Morpholines / Q000494:pharmacology; D009569:Nitric Oxide / Q000378:metabolism*; D020030:Nitric Oxide Donors / Q000494:pharmacology; D009603:Nitroso Compounds / Q000494:pharmacology; D009946:Organothiophosphorus Compounds / Q000494:pharmacology; D010101:Oxygen Consumption / Q000187:drug effects; D051381:Rats; D018160:Receptors, Cytoplasmic and Nuclear / Q000378:metabolism*; D026403:S-Nitrosothiols / Q000378:metabolism*; D000071756:Soluble Guanylyl Cyclase; D013440:Sulfides / Q000378:metabolism*
C529376:GYY 4137; D009025:Morpholines; D020030:Nitric Oxide Donors; D009603:Nitroso Compounds; D009946:Organothiophosphorus Compounds; D018160:Receptors, Cytoplasmic and Nuclear; D026403:S-Nitrosothiols; D013440:Sulfides; C051552:persulfides; D009569:Nitric Oxide; C024942:S-nitrosopenicillamine; D006162:Guanylate Cyclase; D000071756:Soluble Guanylyl Cyclase; D006152:Cyclic GMP
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