Stabilities of Three Key Biological Trisulfides with Implications for Their Roles in the Release of Hydrogen Sulfide and Bioaccumulation of Sulfane Sulfur

[Image: see text] Trisulfides and higher polysulfides are important in the body due to their function as key reservoirs of sulfane sulfur and their rapid reactions to release persulfides. Recent work has shown that persulfides act as powerful antioxidants and release hydrogen sulfide, an emerging gasotransmitter with numerous therapeutic effects. Despite the important role of polysulfides, there is a lack of understanding of their stabilities in aqueous systems. To investigate the reactivity of trisulfides and polysulfides, three key biologically important trisulfides were synthesized from cysteine, glutathione, and N-acetylcysteine, and the tetrasulfide of N-acetylcysteine was synthesized as a representative polysulfide. The stabilities of sulfides were monitored in buffered D(2)O using (1)H NMR spectroscopy under a range of conditions including high temperatures and acidic and alkaline environments. The tri- and tetrasulfides degraded rapidly in the presence of primary and tertiary amines to the corresponding disulfide and elemental sulfur. The half-lives of N-acetylcysteine tri- and tetrasulfides in the presence of butylamine were 53 and 1.5 min, respectively. These results were important because they suggest that tri- and tetrasulfide linkages are short-lived species in vivo due to the abundance of amines in the body. Under basic conditions, cysteine and glutathione trisulfides were unstable due to the deprotonation of the ammonium group, exposing an amine; however, N-acetylcysteine trisulfide was stable at all pH values tested. Hydrogen sulfide release of each polysulfide in the presence of cysteine was quantified using a hydrogen sulfide-sensitive electrode and (1)H NMR spectroscopy.