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

Kimura Y, Toyofuku Y, Koike S, Shibuya N, Nagahara N, Lefer D, Ogasawara Y, Kimura H - Sci Rep (2015)

Bottom Line: We recently found H2Sn in the brain.Purified recombinant 3MST and lysates of COS cells expressing 3MST produced H2S3 from 3 MP, while those expressing defective 3MST mutants did not.The present study provides new insights into the physiology of H2S3 and H2S, as well as novel therapeutic targets for diseases in which these molecules are involved.

View Article: PubMed Central - PubMed

Affiliation: Department of Molecular Pharmacology, National Institute of Neuroscience, National Center of Neurology and Psychiatry, 4-1-1 Ogawahigashi, Kodaira, Tokyo 187-8502, Japan.

ABSTRACT
Hydrogen polysulfides (H2Sn) have a higher number of sulfane sulfur atoms than hydrogen sulfide (H2S), which has various physiological roles. We recently found H2Sn in the brain. H2Sn induced some responses previously attributed to H2S but with much greater potency than H2S. However, the number of sulfur atoms in H2Sn and its producing enzyme were unknown. Here, we detected H2S3 and H2S, 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 H2S3 and H2S 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 H2S3 from 3 MP, while those expressing defective 3MST mutants did not. H2S3 was localized in the cytosol of cells. H2S3 was also produced from H2S by 3MST and rhodanese. H2S2 was identified as a minor H2Sn, and 3 MP did not affect the H2S5 level. The present study provides new insights into the physiology of H2S3 and H2S, as well as novel therapeutic targets for diseases in which these molecules are involved.

No MeSH data available.


Related in: MedlinePlus

Cellular localization of H2S3 produced from 3 MP in COS cells expressing 3MST and in primary neuronal cultures.(a) Localization of H2S3 in COS cells expressing 3MST. Forty-eight hours after transfection with the 3MST cDNA expression plasmid, COS cells were incubated with 50 μM SSP4 for 20 min, and then exposed to 500 μM 3 MP for 10 min. Cells transfected with an empty vector were used as controls. Scale bar = 100 μm. (b) Localization of H2S3 in primary neuronal cultures. Primary neuronal cultures were incubated with 50 μM SSP4, and then exposed to 500 μM 3 MP for 10 min. Scale bar = 10 μm.
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f4: Cellular localization of H2S3 produced from 3 MP in COS cells expressing 3MST and in primary neuronal cultures.(a) Localization of H2S3 in COS cells expressing 3MST. Forty-eight hours after transfection with the 3MST cDNA expression plasmid, COS cells were incubated with 50 μM SSP4 for 20 min, and then exposed to 500 μM 3 MP for 10 min. Cells transfected with an empty vector were used as controls. Scale bar = 100 μm. (b) Localization of H2S3 in primary neuronal cultures. Primary neuronal cultures were incubated with 50 μM SSP4, and then exposed to 500 μM 3 MP for 10 min. Scale bar = 10 μm.

Mentions: We examined the cellular localization of H2S3 produced by 3MST by using COS cells expressing 3MST and loaded with SSP4, a polysulfide-sensitive fluorescence probe25. H2S3 produced de novo from 3 MP incorporated into cells was localized in the cytosol (Fig. 4a). H2S3 was also localized in the cytosol in primary neuronal cultures (Fig. 4b).


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

Kimura Y, Toyofuku Y, Koike S, Shibuya N, Nagahara N, Lefer D, Ogasawara Y, Kimura H - Sci Rep (2015)

Cellular localization of H2S3 produced from 3 MP in COS cells expressing 3MST and in primary neuronal cultures.(a) Localization of H2S3 in COS cells expressing 3MST. Forty-eight hours after transfection with the 3MST cDNA expression plasmid, COS cells were incubated with 50 μM SSP4 for 20 min, and then exposed to 500 μM 3 MP for 10 min. Cells transfected with an empty vector were used as controls. Scale bar = 100 μm. (b) Localization of H2S3 in primary neuronal cultures. Primary neuronal cultures were incubated with 50 μM SSP4, and then exposed to 500 μM 3 MP for 10 min. Scale bar = 10 μm.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4594004&req=5

f4: Cellular localization of H2S3 produced from 3 MP in COS cells expressing 3MST and in primary neuronal cultures.(a) Localization of H2S3 in COS cells expressing 3MST. Forty-eight hours after transfection with the 3MST cDNA expression plasmid, COS cells were incubated with 50 μM SSP4 for 20 min, and then exposed to 500 μM 3 MP for 10 min. Cells transfected with an empty vector were used as controls. Scale bar = 100 μm. (b) Localization of H2S3 in primary neuronal cultures. Primary neuronal cultures were incubated with 50 μM SSP4, and then exposed to 500 μM 3 MP for 10 min. Scale bar = 10 μm.
Mentions: We examined the cellular localization of H2S3 produced by 3MST by using COS cells expressing 3MST and loaded with SSP4, a polysulfide-sensitive fluorescence probe25. H2S3 produced de novo from 3 MP incorporated into cells was localized in the cytosol (Fig. 4a). H2S3 was also localized in the cytosol in primary neuronal cultures (Fig. 4b).

Bottom Line: We recently found H2Sn in the brain.Purified recombinant 3MST and lysates of COS cells expressing 3MST produced H2S3 from 3 MP, while those expressing defective 3MST mutants did not.The present study provides new insights into the physiology of H2S3 and H2S, as well as novel therapeutic targets for diseases in which these molecules are involved.

View Article: PubMed Central - PubMed

Affiliation: Department of Molecular Pharmacology, National Institute of Neuroscience, National Center of Neurology and Psychiatry, 4-1-1 Ogawahigashi, Kodaira, Tokyo 187-8502, Japan.

ABSTRACT
Hydrogen polysulfides (H2Sn) have a higher number of sulfane sulfur atoms than hydrogen sulfide (H2S), which has various physiological roles. We recently found H2Sn in the brain. H2Sn induced some responses previously attributed to H2S but with much greater potency than H2S. However, the number of sulfur atoms in H2Sn and its producing enzyme were unknown. Here, we detected H2S3 and H2S, 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 H2S3 and H2S 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 H2S3 from 3 MP, while those expressing defective 3MST mutants did not. H2S3 was localized in the cytosol of cells. H2S3 was also produced from H2S by 3MST and rhodanese. H2S2 was identified as a minor H2Sn, and 3 MP did not affect the H2S5 level. The present study provides new insights into the physiology of H2S3 and H2S, as well as novel therapeutic targets for diseases in which these molecules are involved.

No MeSH data available.


Related in: MedlinePlus