The Experts below are selected from a list of 630 Experts worldwide ranked by ideXlab platform
Csaba Szabo - One of the best experts on this subject based on the ideXlab platform.
-
novel aryl substituted pyrimidones as inhibitors of 3 Mercaptopyruvate Sulfurtransferase with antiproliferative efficacy in colon cancer
Journal of Medicinal Chemistry, 2021Co-Authors: Marina Bantzi, Fiona Augsburger, Csaba Szabo, Jeremie Loup, Yan Berset, Sofia Vasilakaki, Vassilios Myrianthopoulos, Emmanuel Mikros, Christian G BochetAbstract:The enzyme 3-Mercaptopyruvate Sulfurtransferase (3-MST) is one of the more recently identified mammalian sources of H2S. A recent study identified several novel 3-MST inhibitors with micromolar potency. Among those, (2-[(4-hydroxy-6-methylpyrimidin-2-yl)sulfanyl]-1-(naphthalen-1-yl)ethan-1-one) or HMPSNE was found to be the most potent and selective. We now took the central core of this compound and modified the pyrimidone and the arylketone sides independently. A 63-compound library was synthesized; compounds were tested for H2S generation from recombinant 3-MST in vitro. Active compounds were subsequently tested to elucidate their potency and selectivity. Computer modeling studies have delineated some of the key structural features necessary for binding to the 3-MST's active site. Six novel 3-MST inhibitors were tested in cell-based assays: they exerted inhibitory effects in murine MC38 and CT26 colon cancer cell proliferation; the antiproliferative effect of the compound with the highest potency and best cell-based activity (1b) was also confirmed on the growth of MC38 tumors in mice.
-
role of hydrogen sulfide and 3 Mercaptopyruvate Sulfurtransferase in the regulation of the endoplasmic reticulum stress response in hepatocytes
Biomolecules, 2020Co-Authors: Theodora Panagaki, Elisa B Randi, Csaba SzaboAbstract:It is estimated that over 1.5 billion people suffer from various forms of chronic liver disease worldwide. The emerging prevalence of metabolic syndromes and alcohol misuse, along with the lack of disease-modifying agents for the therapy of many severe liver conditions predicts that chronic liver disease will continue to be a major problem in the future. Better understanding of the underlying pathogenetic mechanisms and identification of potential therapeutic targets remains a priority. Herein, we explored the potential role of the 3-Mercaptopyruvate Sulfurtransferase/hydrogen sulfide (H2S) system in the regulation of the endoplasmic reticulum (ER) stress and of its downstream processes in the immortalized hepatic cell line HepG2 in vitro. ER stress suppressed endogenous H2S levels and pharmacological supplementation of H2S with sodium hydrogen sulfide (NaHS) mitigated many aspects of ER stress, culminating in improved cellular bioenergetics and prevention of autophagic arrest, thereby switching cells’ fate towards survival. Genetic silencing of 3-MST or pharmacological inhibition of the key enzymes involved in hepatocyte H2S biosynthesis exacerbated many readouts related to ER-stress or its downstream functional responses. Our findings implicate the 3-MST/H2S system in the intracellular network that governs proteostasis and ER-stress adaptability in hepatocytes and reinforce the therapeutic potential of pharmacological H2S supplementation.
-
role of 3 Mercaptopyruvate Sulfurtransferase in the regulation of proliferation and cellular bioenergetics in human down syndrome fibroblasts
Biomolecules, 2020Co-Authors: Theodora Panagaki, Elisa B Randi, Csaba SzaboAbstract:Down syndrome (trisomy of human chromosome 21) is a common genetic disorder. Overproduction of the gaseous mediator hydrogen sulfide (H2S) has been implicated in the pathogenesis of neurological and metabolic deficits associated with Down syndrome. Several lines of data indicate that an important enzyme responsible for H2S overproduction in Down syndrome is cystathionine-β-synthase (CBS), an enzyme localized on chromosome 21. The current study explored the possibility that a second H2S-producing enzyme, 3-Mercaptopyruvate Sulfurtransferase (3-MST), may also contribute to the development of functional deficits of Down syndrome cells. Western blotting analysis demonstrated a significantly higher level of 3-MST protein expression in human Down syndrome fibroblasts compared to cells from healthy control individuals; the excess 3-MST was mainly localized to the mitochondrial compartment. Pharmacological inhibition of 3-MST activity improved mitochondrial electron transport and oxidative phosphorylation parameters (but did not affect the suppressed glycolytic parameters) and enhanced cell proliferation in Down syndrome cells (but not in healthy control cells). The findings presented in the current report suggest that in addition to the indisputable role of CBS, H2S produced from 3-MST may also contribute to the development of mitochondrial metabolic and functional impairments in Down syndrome cells.
-
role of 3 Mercaptopyruvate Sulfurtransferase in the regulation of proliferation migration and bioenergetics in murine colon cancer cells
Biomolecules, 2020Co-Authors: Fiona Augsburger, Elisa B Randi, Mathieu Jendly, Kelly Ascencao, Nahzli Dilek, Csaba SzaboAbstract:3-Mercaptopyruvate Sulfurtransferase (3-MST) has emerged as one of the significant sources of biologically active sulfur species in various mammalian cells. The current study was designed to investigate the functional role of 3-MST's catalytic activity in the murine colon cancer cell line CT26. The novel pharmacological 3-MST inhibitor HMPSNE was used to assess cancer cell proliferation, migration and bioenergetics in vitro. Methods included measurements of cell viability (MTT and LDH assays), cell proliferation and in vitro wound healing (IncuCyte) and cellular bioenergetics (Seahorse extracellular flux analysis). 3-MST expression was detected by Western blotting; H2S production was measured by the fluorescent dye AzMC. The results show that CT26 cells express 3-MST protein and mRNA, as well as several enzymes involved in H2S degradation (TST, ETHE1). Pharmacological inhibition of 3-MST concentration-dependently suppressed H2S production and, at 100 and 300 µM, attenuated CT26 proliferation and migration. HMPSNE exerted a bell-shaped effect on several cellular bioenergetic parameters related to oxidative phosphorylation, while other bioenergetic parameters were either unaffected or inhibited at the highest concentration of the inhibitor tested (300 µM). In contrast to 3-MST, the expression of CBS (another H2S producing enzyme which has been previously implicated in the regulation of various biological parameters in other tumor cells) was not detectable in CT26 cells and pharmacological inhibition of CBS exerted no significant effects on CT26 proliferation or bioenergetics. In summary, 3-MST catalytic activity significantly contributes to the regulation of cellular proliferation, migration and bioenergetics in CT26 murine colon cancer cells. The current studies identify 3-MST as the principal source of biologically active H2S in this cell line.
-
3 Mercaptopyruvate Sulfurtransferase supports endothelial cell angiogenesis and bioenergetics
British Journal of Pharmacology, 2020Co-Authors: Armita Abdollahi Govar, Sofia Iris Bibli, Gabor Torő, Peter Szaniszlo, Athanasia Pavlidou, Ketan Thanki, Vicente A Resto, Celia Chao, Mark R Hellmich, Csaba SzaboAbstract:BACKGROUND AND PURPOSE During angiogenesis, quiescent endothelial cells (ECs) are activated by various stimuli to form new blood vessels from pre-existing ones in physiological and pathological conditions. Many research groups have shown that hydrogen sulfide (H2 S), the newest member of the gasotransmitter family, acts as a proangiogenic factor. To date, very little is known about the regulatory role of 3-Mercaptopyruvate Sulfurtransferase (3-MST), an important H2 S-producing enzyme in ECs. The aim of our study was to explore the potential role of 3-MST in human EC bioenergetics, metabolism, and angiogenesis. EXPERIMENTAL APPROACH To assess in vitro angiogenic responses, we used EA.hy926 human vascular ECs subjected to shRNA-mediated 3-MST attenuation and pharmacological inhibition of proliferation, migration, and tube-like network formation. To evaluate bioenergetic parameters, cell respiration, glycolysis, glucose uptake, and mitochondrial/glycolytic ATP production were measured. Finally, global metabolomic profiling was performed to determine the level of 669 metabolic compounds. KEY RESULTS 3-MST-attenuated ECs subjected to shRNA or pharmacological inhibition of 3-MST significantly reduced EC proliferation, migration, and tube-like network formation. 3-MST silencing also suppressed VEGF-induced EC migration. From bioenergetic and metabolic standpoints, 3-MST attenuation decreased mitochondrial respiration and mitochondrial ATP production, increased glucose uptake, and perturbed the entire EC metabolome. CONCLUSION AND IMPLICATIONS 3-MST regulates bioenergetics and morphological angiogenic functions in human ECs. The data presented in the current report support the view that 3-MST pathway may be a potential candidate for therapeutic modulation of angiogenesis. LINKED ARTICLES This article is part of a themed section on Hydrogen Sulfide in Biology & Medicine. To view the other articles in this section visit http://onlinelibrary.wiley.com/doi/10.1111/bph.v177.4/issuetoc.
Noriyuki Nagahara - One of the best experts on this subject based on the ideXlab platform.
-
the h2s generating enzyme 3 Mercaptopyruvate Sulfurtransferase regulates pulmonary vascular smooth muscle cell migration and proliferation but does not impact normal or aberrant lung development
Nitric Oxide, 2021Co-Authors: Noriyuki Nagahara, Ettore Lignelli, Francesco Palumbo, Selahattin Gorkem Bayindir, Istvan Vadasz, Susanne Herold, Werner SeegerAbstract:Abstract Along with nitric oxide (NO), the gasotransmitters carbon monoxide (CO) and hydrogen sulfide (H2S) are emerging as potentially important players in newborn physiology, as mediators of newborn disease, and as new therapeutic modalities. Several recent studies have addressed H2S in particular in animal models of bronchopulmonary dysplasia (BPD), a common complication of preterm birth where oxygen toxicity stunts lung development. In those studies, exogenous H2S attenuated the impact of oxygen toxicity on lung development, and two H2S-generating enzymes were documented to affect pulmonary vascular development. H2S is directly generated endogenously by three enzymes, one of which, 3-Mercaptopyruvate Sulfurtransferase (MPST), has not been studied in the lung. In a hyperoxia-based animal model of BPD, oxygen exposure deregulated MPST expression during post-natal lung development, where MPST was localized to the smooth muscle layer of the pulmonary vessels in developing lungs. siRNA-mediated abrogation of MPST expression in human pulmonary artery smooth muscle cells in vitro limited baseline cell migration and cell proliferation, without affecting apoptosis or cell viability. In vivo, MPST was dispensable for normal lung development in Mpst-/-mice, and MPST did not contribute to stunted lung development driven by hyperoxia exposure, assessed by design-based stereology. These data demonstrate novel roles for MPST in pulmonary vascular smooth muscle cell physiology. The potential caveats of using Mpst-/- mice to study normal and aberrant lung development are also discussed, highlighting the possible confounding, compensatory effects of other H2S-generating enzymes that are present alongside MPST in the smooth muscle compartment of developing pulmonary vessels.
-
Activation of 3-Mercaptopyruvate Sulfurtransferase by Glutaredoxin Reducing System.
Biomolecules, 2020Co-Authors: Noriyuki NagaharaAbstract:Glutaredoxin (EC 1.15–1.21) is known as an oxidoreductase that protects cysteine residues within proteins against oxidative stress. Glutaredoxin catalyzes an electron transfer reaction that donates an electron to substrate proteins in the reducing system composed of glutaredoxin, glutathione, glutathione reductase, and nicotinamide-adenine dinucleotide phosphate (reduced form). 3-Mercaptopyruvate Sulfurtransferase (EC 2.8.1.2) is a cysteine enzyme that catalyzes transsulfuration, and glutaredoxin activates 3-Mercaptopyruvate Sulfurtransferase in the reducing system. Interestingly, even when glutathione or glutathione reductase was absent, 3-Mercaptopyruvate Sulfurtransferase activity increased, probably because reduced glutaredoxin was partly present and able to activate 3-Mercaptopyruvate Sulfurtransferase until depletion. A study using mutant Escherichia coli glutaredoxin1 (Cys14 is the binding site of glutathione and was replaced with a Ser residue) confirmed these results. Some inconsistency was noted, and glutaredoxin with higher redox potential than either 3-Mercaptopyruvate Sulfurtransferase or glutathione reduced 3-Mercaptopyruvate Sulfurtransferase. However, electron-transfer enzymatically proceeded from glutaredoxin to 3-Mercaptopyruvate Sulfurtransferase.
-
the protective role of the 3 Mercaptopyruvate Sulfurtransferase 3 mst hydrogen sulfide h2s pathway against experimental osteoarthritis
Arthritis Research & Therapy, 2020Co-Authors: Sonia Nasi, Noriyuki Nagahara, Athanasia Chatzianastasiou, Andreas Papapetropoulos, Giuseppe Cirino, Driss Ehirchiou, J Bertrand, Nathalie BussoAbstract:Osteoarthritis (OA) is characterized by the formation and deposition of calcium-containing crystals in joint tissues, but the underlying mechanisms are poorly understood. The gasotransmitter hydrogen sulfide (H2S) has been implicated in mineralization but has never been studied in OA. Here, we investigated the role of the H2S-producing enzyme 3-Mercaptopyruvate Sulfurtransferase (3-MST) in cartilage calcification and OA development. 3-MST expression was analyzed in cartilage from patients with different OA degrees, and in cartilage stimulated with hydroxyapatite (HA) crystals. The modulation of 3-MST expression in vivo was studied in the meniscectomy (MNX) model of murine OA, by comparing sham-operated to MNX knee cartilage. The role of 3-MST was investigated by quantifying joint calcification and cartilage degradation in WT and 3-MST−/− meniscectomized knees. Chondrocyte mineralization in vitro was measured in WT and 3-MST−/− cells. Finally, the effect of oxidative stress on 3-MST expression and chondrocyte mineralization was investigated. 3-MST expression in human cartilage negatively correlated with calcification and OA severity, and diminished upon HA stimulation. In accordance, cartilage from menisectomized OA knees revealed decreased 3-MST if compared to sham-operated healthy knees. Moreover, 3-MST−/− mice showed exacerbated joint calcification and OA severity if compared to WT mice. In vitro, genetic or pharmacologic inhibition of 3-MST in chondrocytes resulted in enhanced mineralization and IL-6 secretion. Finally, oxidative stress decreased 3-MST expression and increased chondrocyte mineralization, maybe via induction of pro-mineralizing genes. 3-MST-generated H2S protects against joint calcification and experimental OA. Enhancing H2S production in chondrocytes may represent a potential disease modifier to treat OA.
-
abstract 631 complementary roles for 3 Mercaptopyruvate Sulfurtransferase and cystathionine γ lyase in angiogenesis
Circulation Research, 2019Co-Authors: Athanasia Pavlidou, Noriyuki Nagahara, Csaba Szabo, Sofia Iris Bibli, Katalin Modis, Ingrid Fleming, Andreas PapapetropoulosAbstract:Introduction: Hydrogen sulfide (H2S) is a signaling molecule with important actions in the cardiovascular system. The major endogenous sources for H2S in the endothelium are cystathionine γ-lyase (...
-
Novel Characterization of Antioxidant Enzyme, 3-Mercaptopyruvate Sulfurtransferase-Knockout Mice: Overexpression of the Evolutionarily-Related Enzyme Rhodanese
Antioxidants, 2019Co-Authors: Noriyuki Nagahara, Mio Tanaka, Yukichi Tanaka, Takaaki ItoAbstract:The antioxidant enzyme, 3-Mercaptopyruvate Sulfurtransferase (MST, EC 2.8.1.2) is localized in the cytosol and mitochondria, while the evolutionarily-related enzyme, rhodanese (thiosulfate Sulfurtransferase, TST, EC 2.8.1.1) is localized in the mitochondria. Recently, both enzymes have been shown to produce hydrogen sulfide and polysulfide. Subcellular fractionation of liver mitochondria revealed that the TST activity ratio of MST-knockout (KO)/wild-type mice was approximately 2.5; MST activity was detected only in wild-type mice, as expected. The ratio of TST mRNA expression of KO/wild-type mice, as measured by real-time quantitative polymerase chain reaction analysis, was approximately 3.3. It is concluded that TST is overexpressed in MST-KO mice.
Hideo Kimura - One of the best experts on this subject based on the ideXlab platform.
-
signaling by hydrogen sulfide h2s and polysulfides h2sn in the central nervous system
Neurochemistry International, 2019Co-Authors: Hideo KimuraAbstract:Hydrogen sulfide (H2S) is a signaling molecule used to modify neuronal transmission, regulate vascular tone, protect tissues from oxidative stress, sense oxygen, and generate ATP. Hydrogen polysulfides (H2Sn) have recently been identified as signaling molecules that mediate the activation of ion channels, regulation of tumor growth, and the transcriptional regulation of oxidative stress; some of which were previously ascribed to H2S. Cystathionine β-synthetase (CBS), cystathionine γ-lyase (CSE), and 3-Mercaptopyruvate Sulfurtransferase (3MST) are known as H2S-producing enzymes. 3MST also produces H2Sn and other persulfurated molecules such as cysteine persulfide, glutathione persulfide, and persulfurated proteins. The chemical interaction of H2S and nitric oxide (NO) also produces H2Sn, which may be the mechanism underlying the synergistic effect of H2S and NO that was initially reported on vascular relaxation. H2Sn and other persulfurated molecules elicit their effect via S-sulfuration (S-sulfhydration) of specific cysteine residues of the target proteins. This review article focuses on the production and roles of H2Sn as well as H2S in the central nervous system.
-
alternative pathway of h2s and polysulfides production from sulfurated catalytic cysteine of reaction intermediates of 3 Mercaptopyruvate Sulfurtransferase
Biochemical and Biophysical Research Communications, 2018Co-Authors: Noriyuki Nagahara, Hideo Kimura, Shin Koike, Takashi Nirasawa, Yuki OgasawaraAbstract:Abstract It has been known that hydrogen sulfide and/or polysulfides are produced from a (poly)sulfurated sulfur-acceptor substrate of 3-Mercaptopyruvate Sulfurtransferase (MST) via thioredoxin (Trx) reduction in vitro. In this study, we used thiosulfate as the donor substrate and the catalytic reaction was terminated on the formation of a persulfide or polysulfides. We can present alternative pathway of production of hydrogen sulfide and/or polysulfides from (poly)sulfurated catalytic-site cysteine of reaction intermediates of MST via Trx reduction. Matrix-assisted laser desorption ionization tandem time-of-flight mass spectrometric analysis revealed that after prolonged incubation of MST with thiosulfate, a trisulfide adduct becomes predominant at the sulfurated catalytic-site cysteine. When these adducts were reduced by Trx with reducing system (MST:Escherichia coli Trx:E. coli Trx reductase:NADPH = 1:5:0.02:12.5 molar ratio), liquid chromatography with tandem mass spectrometric analysis for monobromobimane-derivatized H2Sn revealed that H2S2 first appeared, and then H2S and H2S3 did later. The results were confirmed by high-performance liquid chromatography-fluorescence analysis.
-
3 Mercaptopyruvate Sulfurtransferase produces potential redox regulators cysteine and glutathione persulfide cys ssh and gssh together with signaling molecules h 2 s 2 h 2 s 3 and h 2 s
Scientific Reports, 2017Co-Authors: Yuka Kimura, Norihiro Shibuya, David J. Lefer, Yuki Ogasawara, Shin Koike, Hideo KimuraAbstract:Cysteine-persulfide (Cys-SSH) is a cysteine whose sulfhydryl group is covalently bound to sulfur (sulfane sulfur). Cys-SSH and its glutathione (GSH) counterpart (GSSH) have been recognized as redox regulators, some of which were previously ascribed to cysteine and GSH. However, the production of Cys-SSH and GSSH is not well understood. Here, we show that 3-Mercaptopyruvate Sulfurtransferase (3MST) produces Cys-SSH and GSSH together with the potential signaling molecules hydrogen per- and tri-sulfide (H2S2 and H2S3). Cys-SSH and GSSH are produced in the brain of wild-type mice but not in those of 3MST-KO mice. The levels of total persulfurated species in the brain of 3MST-KO mice are less than 50% of that in the brain of wild-type mice. Purified recombinant 3MST and lysates of COS cells expressing 3MST showed that Cys-SSH and GSSH were produced in the presence of physiological concentrations of cysteine and glutathione, while those with longer sulfur chains, Cys-SSnH and GSSnH, were produced in the presence of lower than physiological concentrations of cysteine and glutathione. The present study provides new insights into the production and physiological roles of these persulfurated species as well as the therapeutic targets for diseases in which these molecules are involved.
-
identification of h2s3 and h2s produced by 3 Mercaptopyruvate Sulfurtransferase in the brain
Scientific Reports, 2015Co-Authors: Yuka Kimura, Norihiro Shibuya, Noriyuki Nagahara, David J. Lefer, Yuki Ogasawara, Yukiko Toyofuku, Shin Koike, Hideo KimuraAbstract: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.
-
hydrogen sulfide and polysulfides as signaling molecules
Nitric Oxide, 2015Co-Authors: Hideo KimuraAbstract:Hydrogen sulfide (H2S) is a familiar toxic gas that smells of rotten eggs. After the identification of endogenous H2S in the mammalian brain two decades ago, studies of this molecule uncovered physiological roles in processes such as neuromodulation, vascular tone regulation, cytoprotection against oxidative stress, angiogenesis, anti-inflammation, and oxygen sensing. Enzymes that produce H2S, such as cystathionine β-synthase, cystathionine γ-lyase, and 3-Mercaptopyruvate Sulfurtransferase have been studied intensively and well characterized. Polysulfides, which have a higher number of inner sulfur atoms than that in H2S, were recently identified as potential signaling molecules that can activate ion channels, transcription factors, and tumor suppressors with greater potency than that of H2S. This article focuses on our contribution to the discovery of these molecules and their metabolic pathways and mechanisms of action.
Andreas Papapetropoulos - One of the best experts on this subject based on the ideXlab platform.
-
the protective role of the 3 Mercaptopyruvate Sulfurtransferase 3 mst hydrogen sulfide h2s pathway against experimental osteoarthritis
Arthritis Research & Therapy, 2020Co-Authors: Sonia Nasi, Noriyuki Nagahara, Athanasia Chatzianastasiou, Andreas Papapetropoulos, Giuseppe Cirino, Driss Ehirchiou, J Bertrand, Nathalie BussoAbstract:Osteoarthritis (OA) is characterized by the formation and deposition of calcium-containing crystals in joint tissues, but the underlying mechanisms are poorly understood. The gasotransmitter hydrogen sulfide (H2S) has been implicated in mineralization but has never been studied in OA. Here, we investigated the role of the H2S-producing enzyme 3-Mercaptopyruvate Sulfurtransferase (3-MST) in cartilage calcification and OA development. 3-MST expression was analyzed in cartilage from patients with different OA degrees, and in cartilage stimulated with hydroxyapatite (HA) crystals. The modulation of 3-MST expression in vivo was studied in the meniscectomy (MNX) model of murine OA, by comparing sham-operated to MNX knee cartilage. The role of 3-MST was investigated by quantifying joint calcification and cartilage degradation in WT and 3-MST−/− meniscectomized knees. Chondrocyte mineralization in vitro was measured in WT and 3-MST−/− cells. Finally, the effect of oxidative stress on 3-MST expression and chondrocyte mineralization was investigated. 3-MST expression in human cartilage negatively correlated with calcification and OA severity, and diminished upon HA stimulation. In accordance, cartilage from menisectomized OA knees revealed decreased 3-MST if compared to sham-operated healthy knees. Moreover, 3-MST−/− mice showed exacerbated joint calcification and OA severity if compared to WT mice. In vitro, genetic or pharmacologic inhibition of 3-MST in chondrocytes resulted in enhanced mineralization and IL-6 secretion. Finally, oxidative stress decreased 3-MST expression and increased chondrocyte mineralization, maybe via induction of pro-mineralizing genes. 3-MST-generated H2S protects against joint calcification and experimental OA. Enhancing H2S production in chondrocytes may represent a potential disease modifier to treat OA.
-
abstract 631 complementary roles for 3 Mercaptopyruvate Sulfurtransferase and cystathionine γ lyase in angiogenesis
Circulation Research, 2019Co-Authors: Athanasia Pavlidou, Noriyuki Nagahara, Csaba Szabo, Sofia Iris Bibli, Katalin Modis, Ingrid Fleming, Andreas PapapetropoulosAbstract:Introduction: Hydrogen sulfide (H2S) is a signaling molecule with important actions in the cardiovascular system. The major endogenous sources for H2S in the endothelium are cystathionine γ-lyase (...
-
Mercaptopyruvate acts as endogenous vasodilator independently of 3 Mercaptopyruvate Sulfurtransferase activity
Nitric Oxide, 2018Co-Authors: Emma Mitidieri, Noriyuki Nagahara, Antonia Katsouda, Teresa Tramontano, Danila Gurgone, Valentina Citi, Vincenzo Calderone, Vincenzo Brancaleone, Andreas Papapetropoulos, Giuseppe CirinoAbstract:Abstract Hydrogen sulfide (H2S) is produced by the action of cystathionine-β-synthase (CBS), cystathionine-γ-lyase (CSE) or 3-Mercaptopyruvate Sulfurtransferase (3-MST). 3-MST converts 3-Mercaptopyruvate (MPT) to H2S and pyruvate. H2S is recognized as an endogenous gaseous mediator with multiple regulatory roles in mammalian cells and organisms. In the present study we demonstrate that MPT, the endogenous substrate of 3-MST, acts also as endogenous H2S donor. Colorimetric, amperometric and fluorescence based assays demonstrated that MPT releases H2S in vitro in an enzyme-independent manner. A functional study was performed on aortic rings harvested from C57BL/6 (WT) or 3-MST-knockout (3-MST−/−) mice with and without endothelium. MPT relaxed mouse aortic rings in endothelium-independent manner and at the same extent in both WT and 3-MST−/− mice. N5-(1-Iminoethyl)- l -ornithine dihydrochloride (L-NIO, an inhibitor of endothelial nitric oxide synthase) as well as 1H-[1,2,4]oxadiazolo [4,3-a]quinoxalin-1-one (ODQ, a soluble guanylyl cyclase inhibitor) did not affect MPT relaxant action. Conversely, hemoglobin (as H2S scavenger), as well as glybenclamide (an ATP-dependent potassium channel blocker) markedly reduced MPT-induced relaxation. The functional data clearly confirmed a non enzymatic vascular effect of MPT. In conclusion, MPT acts also as an endogenous H2S donor and not only as 3-MST substrate. MPT could, thus, be further investigated as a means to increase H2S in conditions where H2S bioavailability is reduced such as hypertension, coronary artery disease, diabetes or urogenital tract disease.
-
regulation of vascular tone angiogenesis and cellular bioenergetics by the 3 Mercaptopyruvate Sulfurtransferase h2s pathway functional impairment by hyperglycemia and restoration by dl α lipoic acid
Molecular Medicine, 2015Co-Authors: Ciro Coletta, Andreas Papapetropoulos, Katalin Modis, Bartosz Szczesny, Attila Brunyánszki, Gábor Oláh, Kazunori Yanagi, Akbar Ahmad, Ester Rios, Csaba SzaboAbstract:Hydrogen sulfide (H2S), as a reducing agent and an antioxidant molecule, exerts protective effects against hyperglycemic stress in the vascular endothelium. The mitochondrial enzyme 3-Mercaptopyruvate Sulfurtransferase (3-MST) is an important biological source of H2S. We have recently demonstrated that 3-MST activity is inhibited by oxidative stress in vitro and speculated that this may have an adverse effect on cellular homeostasis. In the current study, given the importance of H2S as a vasorelaxant, angiogenesis stimulator and cellular bioenergetic mediator, we first determined whether the 3-MST/H2S system plays a physiological regulatory role in endothelial cells. Next, we tested whether a dysfunction of this pathway develops during the development of hyperglycemia and μmol/L to diabetes-associated vascular complications. Intraperitoneal (IP) 3-MP (1 mg/kg) raised plasma H2S levels in rats. 3-MP (10 1 mmol/L) promoted angiogenesis in vitro in bEnd3 microvascular endothelial cells and in vivo in a Matrigel assay in mice (0.3–1 mg/kg). In vitro studies with bEnd3 cell homogenates demonstrated that the 3-MP-induced increases in H2S production depended on enzymatic activity, although at higher concentrations (1–3 mmol/L) there was also evidence for an additional nonenzymatic H2S production by 3-MP. In vivo, 3-MP facilitated wound healing in rats, induced the relaxation of dermal microvessels and increased mitochondrial bioenergetic function. In vitro hyperglycemia or in vivo streptozotocin diabetes impaired angiogenesis, attenuated mitochondrial function and delayed wound healing; all of these responses were associated with an impairment of the proangiogenic and bioenergetic effects of 3-MP. The antioxidants dl-α-lipoic acid (LA) in vivo, or dihydrolipoic acid (DHLA) in vitro restored the ability of 3-MP to stimulate angiogenesis, cellular bioenergetics and wound healing in hyperglycemia and diabetes. We conclude that diabetes leads to an impairment of the 3-MST/H2S pathway, and speculate that this may contribute to the pathogenesis of hyperglycemic endothelial cell dysfunction. We also suggest that therapy with H2S donors, or treatment with the combination of 3-MP and lipoic acid may be beneficial in improving angiogenesis and bioenergetics in hyperglycemia.
-
Role of the cystathionine gamma lyase/hydrogen sulfide pathway in human melanoma progression
'Wiley', 2015Co-Authors: Panza Elisabetta, Andreas Papapetropoulos, Paola De Cicco, Chiara Armogida, Giosue Scognamiglio, Vincenzo Gigantino, Gerardo Botti, Domenico Germano, Maria Napolitano, Bucci MariarosariaAbstract:In humans, two main metabolic enzymes synthesize hydrogen sulfide (H2 S): cystathionine γ lyase (CSE) and cystathionine β synthase (CBS). A third enzyme, 3-Mercaptopyruvate Sulfurtransferase (3-MST), synthesizes H2 S in the presence of the substrate 3-Mercaptopyruvate (3-MP). The immunohistochemistry analysis performed on human melanoma samples demonstrated that CSE expression was highest in primary tumors, decreased in the metastatic lesions and was almost silent in non-lymph node metastases. The primary role played by CSE was confirmed by the finding that the overexpression of CSE induced spontaneous apoptosis of human melanoma cells. The same effect was achieved using different H2 S donors, the most active of which was diallyl trisulfide (DATS). The main pro-apoptotic mechanisms involved were suppression of nuclear factor-κB activity and inhibition of AKT and extracellular signal-regulated kinase pathways. A proof of concept was obtained in vivo using a murine melanoma model. In fact, either l-cysteine, the CSE substrate, or DATS inhibited tumor growth in mice. In conclusion, we have determined that the l-cysteine/CSE/H2 S pathway is involved in melanoma progression
Norihiro Shibuya - One of the best experts on this subject based on the ideXlab platform.
-
production of h 2 s h 2 s n and persulfide species cysssh and gssh by 3 Mercaptopyruvate Sulfurtransferase
Folia Pharmacologica Japonica, 2018Co-Authors: Norihiro ShibuyaAbstract:Accumulating evidence shows that hydrogen sulfide (H2S) has physiological roles in various tissues and organs, including the regulation of neuronal activity, vascular tension, a release of insulin, and protection of the heart, kidney, and brain from ischemic insult. H2S is produced from l-cysteine by pyridoxal 5'-phosphate (PLP)-dependent enzymes, cystathionine β-synthase (CBS) and cystathionine γ-lyase (CSE). 3-Mercaptopyruvate Sulfurtransferase (3MST) is the third H2S-producing enzyme, and its substrate 3-Mercaptopyruvate (3MP) is provided from l-cysteine and α-ketoglutarate (α-KG) by a PLP-dependent cysteine aminotransferase (CAT). An additional pathway for the production of H2S from d-cysteine metabolized by d-amino acid oxidase (DAO) together with 3MST has been identified. Recent studies have shown that hydrogen polysulfides (H2Sn) have been found to stimulate transient receptor potential ankyrin1 (TRPA1) channel, much more potently than does H2S. 3MST produces cysteine-persulfide (CysSSH) and its glutathione counterpart (GSSH), potential redox regulators, together with the potential signaling molecules H2Sn. In addition, the interaction between H2S and nitric oxide (NO) also generates H2Sn. These observations provide new insights into the production and physiological roles of these molecules.
-
3 Mercaptopyruvate Sulfurtransferase produces potential redox regulators cysteine and glutathione persulfide cys ssh and gssh together with signaling molecules h 2 s 2 h 2 s 3 and h 2 s
Scientific Reports, 2017Co-Authors: Yuka Kimura, Norihiro Shibuya, David J. Lefer, Yuki Ogasawara, Shin Koike, Hideo KimuraAbstract:Cysteine-persulfide (Cys-SSH) is a cysteine whose sulfhydryl group is covalently bound to sulfur (sulfane sulfur). Cys-SSH and its glutathione (GSH) counterpart (GSSH) have been recognized as redox regulators, some of which were previously ascribed to cysteine and GSH. However, the production of Cys-SSH and GSSH is not well understood. Here, we show that 3-Mercaptopyruvate Sulfurtransferase (3MST) produces Cys-SSH and GSSH together with the potential signaling molecules hydrogen per- and tri-sulfide (H2S2 and H2S3). Cys-SSH and GSSH are produced in the brain of wild-type mice but not in those of 3MST-KO mice. The levels of total persulfurated species in the brain of 3MST-KO mice are less than 50% of that in the brain of wild-type mice. Purified recombinant 3MST and lysates of COS cells expressing 3MST showed that Cys-SSH and GSSH were produced in the presence of physiological concentrations of cysteine and glutathione, while those with longer sulfur chains, Cys-SSnH and GSSnH, were produced in the presence of lower than physiological concentrations of cysteine and glutathione. The present study provides new insights into the production and physiological roles of these persulfurated species as well as the therapeutic targets for diseases in which these molecules are involved.
-
discovery and mechanistic characterization of selective inhibitors of h2s producing enzyme 3 Mercaptopyruvate Sulfurtransferase 3mst targeting active site cysteine persulfide
Scientific Reports, 2017Co-Authors: Kenjiro Hanaoka, Norihiro Shibuya, Yusuke Suwanai, Kazuhito Shimamoto, Kiyoshi Sasakura, Sachiko Tomafukai, Yoko Takano, Takuya Terai, Toru KomatsuAbstract:Very recent studies indicate that sulfur atoms with oxidation state 0 or −1, called sulfane sulfurs, are the actual mediators of some physiological processes previously considered to be regulated by hydrogen sulfide (H2S). 3-Mercaptopyruvate Sulfurtransferase (3MST), one of three H2S-producing enzymes, was also recently shown to produce sulfane sulfur (H2Sn). Here, we report the discovery of several potent 3MST inhibitors by means of high-throughput screening (HTS) of a large chemical library (174,118 compounds) with our H2S-selective fluorescent probe, HSip-1. Most of the identified inhibitors had similar aromatic ring-carbonyl-S-pyrimidone structures. Among them, compound 3 showed very high selectivity for 3MST over other H2S/sulfane sulfur-producing enzymes and rhodanese. The X-ray crystal structures of 3MST complexes with two of the inhibitors revealed that their target is a persulfurated cysteine residue located in the active site of 3MST. Precise theoretical calculations indicated the presence of a strong long-range electrostatic interaction between the persulfur anion of the persulfurated cysteine residue and the positively charged carbonyl carbon of the pyrimidone moiety of the inhibitor. Our results also provide the experimental support for the idea that the 3MST-catalyzed reaction with 3-Mercaptopyruvate proceeds via a ping-pong mechanism.
-
discovery and mechanistic characterization of selective inhibitors of h 2 s producing enzyme 3 Mercaptopyruvate Sulfurtransferase 3mst targeting active site cysteine persulfide
Scientific Reports, 2017Co-Authors: Kenjiro Hanaoka, Norihiro Shibuya, Yusuke Suwanai, Kazuhito Shimamoto, Kiyoshi Sasakura, Sachiko Tomafukai, Yoko Takano, Takuya Terai, Toru KomatsuAbstract:Very recent studies indicate that sulfur atoms with oxidation state 0 or -1, called sulfane sulfurs, are the actual mediators of some physiological processes previously considered to be regulated by hydrogen sulfide (H2S). 3-Mercaptopyruvate Sulfurtransferase (3MST), one of three H2S-producing enzymes, was also recently shown to produce sulfane sulfur (H2Sn). Here, we report the discovery of several potent 3MST inhibitors by means of high-throughput screening (HTS) of a large chemical library (174,118 compounds) with our H2S-selective fluorescent probe, HSip-1. Most of the identified inhibitors had similar aromatic ring-carbonyl-S-pyrimidone structures. Among them, compound 3 showed very high selectivity for 3MST over other H2S/sulfane sulfur-producing enzymes and rhodanese. The X-ray crystal structures of 3MST complexes with two of the inhibitors revealed that their target is a persulfurated cysteine residue located in the active site of 3MST. Precise theoretical calculations indicated the presence of a strong long-range electrostatic interaction between the persulfur anion of the persulfurated cysteine residue and the positively charged carbonyl carbon of the pyrimidone moiety of the inhibitor. Our results also provide the experimental support for the idea that the 3MST-catalyzed reaction with 3-Mercaptopyruvate proceeds via a ping-pong mechanism.
-
identification of h2s3 and h2s produced by 3 Mercaptopyruvate Sulfurtransferase in the brain
Scientific Reports, 2015Co-Authors: Yuka Kimura, Norihiro Shibuya, Noriyuki Nagahara, David J. Lefer, Yuki Ogasawara, Yukiko Toyofuku, Shin Koike, Hideo KimuraAbstract: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.