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Hydrogen polysulfides (H 2 S n ) have a higher number of sulfane sulfur atoms than hydrogen sulfide (H 2 S), which has various physiological roles. We recently found H 2 S n  in the brain. H 2 S n  induced some responses previously attributed to H 2 S but with much greater potency than H 2 S. However, the number of sulfur atoms in H 2 S n  and its producing enzyme were unknown. Here, we detected H 2 S 3  and H 2 S, which were produced from 3-mercaptopyruvate (3 MP) by 3-mercaptopyruvate sulfurtransferase (3MST), in the brain. High performance liquid chromatography with fluorescence detection (LC-FL) and tandem mass spectrometry (LC-MS/MS) analyses showed that H 2 S 3  and H 2 S were produced from 3 MP in the brain cells of wild-type mice but not 3MST knockout (3MST-KO) mice. Purified recombinant 3MST and lysates of COS cells expressing 3MST produced H 2 S 3  from 3 MP, while those expressing defective 3MST mutants did not. H 2 S 3  was localized in the cytosol of cells. H 2 S 3  was also produced from H 2 S by 3MST and rhodanese. H 2 S 2  was identified as a minor H 2 S n , and 3 MP did not affect the H 2 S 5  level. The present study provides new insights into the physiology of H 2 S 3  and H 2 S, as well as novel therapeutic targets for diseases in which these molecules are involved.

Identification of H2S3 and H2S produced by 3-mercaptopyruvate sulfurtransferase in the brain