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Production of H2S5 from H2S (a) H2S5 production from Na2S with lysates of COS cells expressing 3MST or rhodanese as a source of the enzymes. Control: lysates of cells transfected with an empty vector. (b) Production of H2S5 in whole cells prepared from brains of wild-type (open bar) and 3MST-KO mice (filled bar) exposed to 500 μM Na2S for 15 min. Distilled water was applied as a control. A H2S5 standard was not available. For this reasons, relative values are shown for H2S5. *p < 0.05. All data represent the mean ± standard error of the mean (SEM) of at least three experiments.
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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 H2S n and its producing enzyme were...
Citations
... Although CSE levels are very low in the brain, it plays an important role under pathological conditions such as Alzheimer's disease and Huntington's disease [7][8][9]. 3MST produces not only H 2 S but also polysulfides, including H 2 S n [10][11][12]. Cysteine aminotransferase (CAT) metabolizes Lcysteine and α-ketoglutarate to produce 3-mercaptopyruvate (3MP), which is the substrate of 3MST. 3MP is also produced from D-cysteine by D-amino acid oxydase [13]. ...
... Endogenous H 2 S n have been identified in the brain [11,48]. 3MST produces H 2 S n and other S-sulfurated molecules, such as cysteine persulfide (Cys-SSH), glutathione persulfide (GSSH), and S-sulfurated cysteine residues of proteins (P-SSH) [12,71]. Two possible mechanisms have been proposed. ...
... Two possible mechanisms have been proposed. Sulfur is transferred from S-sulfurated active cysteine residues of 3MST to the acceptor molecules such as H 2 S, cysteine, glutathione, and cysteine residues, to produce corresponding S-sulfurated molecules ( Figure 4a) [12,71]. Alternatively, 3MST produces H 2 S 2 , which reacts with cysteine, glutathione, and cysteine residues to generate their S-sulfurated molecules ( Figure 4b) [12,71]. ...
Hydrogen sulfide (H2S) and polysulfides (H2Sn, n ≥ 2) produced by enzymes play a role as signalling molecules regulating neurotransmission, vascular tone, cytoprotection, inflammation, oxygen sensing, and energy formation. H2Sn, which have additional sulfur atoms to H2S, and other S-sulfurated molecules such as cysteine persulfide and S-sulfurated cysteine residues of proteins, are produced by enzymes including 3-mercaptopyruvate sulfurtransferase (3MST). H2Sn are also generated by the chemical interaction of H2S with NO, or to a lesser extent with H2O2. S-sulfuration (S-sulfhydration) has been proposed as a mode of action of H2S and H2Sn to regulate the activity of target molecules. Recently, we found that H2S/H2S2 regulate the release of neurotransmitters, such as GABA, glutamate, and D-serine, a co-agonist of N-methyl-D-aspartate (NMDA) receptors. H2S facilitates the induction of hippocampal long-term potentiation, a synaptic model of memory formation, by enhancing the activity of NMDA receptors, while H2S2 achieves this by activating transient receptor potential ankyrin 1 (TRPA1) channels in astrocytes, potentially leading to the activation of nearby neurons. The recent findings show the other aspects of TRPA1 channels—that is, the regulation of the levels of sulfur-containing molecules and their metabolizing enzymes. Disturbance of the signalling by H2S/H2Sn has been demonstrated to be involved in various diseases, including cognitive and psychiatric diseases. The physiological and pathophysiological roles of these molecules will be discussed.
... The cell growth was determined by the OD 600 using an ultraviolet spectrophotometer (Xinmao Instrument, Shanghai, China). For the determination of H 2 S, samples were derivatized by mBBr and analyzed using the high-performance liquid chromatography (HPLC, Ultimate 3000) equipped with a fluorescence detector and C18 reverse phase column according to the previous method of Kimura et al. (66). The intracellular sulfane sulfur was measured using Sulfane sulfur probe 4 (SSP4) following the reported method (15). ...
Sulfane sulfur, a collective term for hydrogen polysulfide and organic persulfide, often damages cells at high concentrations. Cells can regulate intracellular sulfane sulfur levels through specific mechanisms, but these mechanisms are unclear in Corynebacterium glutamicum . OxyR is a transcription factor capable of sensing oxidative stress and is also responsive to sulfane sulfur. In this study, we found that OxyR functioned directly in regulating sulfane sulfur in C. glutamicum . OxyR binds to the promoter of katA and nrdH and regulates its expression, as revealed via in vitro electrophoretic mobility shift assay analysis, real-time quantitative PCR, and reporting systems. Overexpression of katA and nrdH reduced intracellular sulfane sulfur levels by over 30% and 20% in C. glutamicum , respectively. RNA-sequencing analysis showed that the lack of OxyR downregulated the expression of sulfur assimilation pathway genes and/or sulfur transcription factors, which may reduce the rate of sulfur assimilation. In addition, OxyR also affected the biosynthesis of L-cysteine in C. glutamicum . OxyR overexpression strain Cg-2 accumulated 183 mg/L of L-cysteine, increased by approximately 30% compared with the control (142 mg/L). In summary, OxyR not only regulated sulfane sulfur levels by controlling the expression of katA and nrdH in C. glutamicum but also facilitated the sulfur assimilation and L-cysteine synthesis pathways, providing a potential target for constructing robust cell factories of sulfur-containing amino acids and their derivatives.
IMPORTANCE
C. glutamicum is an important industrial microorganism used to produce various amino acids. In the production of sulfur-containing amino acids, cells inevitably accumulate a large amount of sulfane sulfur. However, few studies have focused on sulfane sulfur removal in C. glutamicum . In this study, we not only revealed the regulatory mechanism of OxyR on intracellular sulfane sulfur removal but also explored the effects of OxyR on the sulfur assimilation and L-cysteine synthesis pathways in C. glutamicum . This is the first study on the removal of sulfane sulfur in C. glutamicum . These results contribute to the understanding of sulfur regulatory mechanisms and may aid in the future optimization of C. glutamicum for biosynthesis of sulfur-containing amino acids.
... H 2 S is a reactive molecule and toxic in the higher concentrations; therefore, under physiological conditions, the level of free H 2 S is tightly controlled and kept at a low level, while its excess is stored as bioavailable sulfur, like bound sulfane sulfur. This kind of reactive sulfur includes mainly persulfides (RSSH), polysulfides (RS n R, n > 2), and inorganic polysulfides (H 2 S n ), which release H 2 S under reducing conditions [35]. increased locomotor activity in the BSO + GBR 12909induced rat model of schizophrenia [13,14]. ...
Preclinical and clinical studies have shown that the antipsychotic drug aripiprazole and the antioxidant N‐acetylcysteine have unique biological properties. The aim of the study was to investigate, in a rat model of schizophrenia, the effects of chronic administration of these drugs on schizophrenia‐like behaviors and anaerobic cysteine metabolism in the hippocampus (HIP). The schizophrenia‐type changes were induced in Sprague–Dawley rats by repeated administration of the glutathione synthesis inhibitor l‐butionine‐(S,R)‐sulfoximine in combination with the dopamine reuptake inhibitor GBR 12909 in the early postnatal period. Adult model rats were chronically treated with aripiprazole (0.3 mg·kg⁻¹, i.p.) or N‐acetylcysteine (30 mg·kg⁻¹, orally), and their effects on schizophrenia‐like behaviors were assessed using the social interaction test and novel object recognition test. In the HIP, the level of anaerobic cysteine metabolites, H2S, and bound sulfane sulfur were determined by a fluorescence method, while the expression of H2S‐synthetizing enzymes: cystathionine β‐synthase (CBS) and mercaptopyruvate sulfurtransferase (MST) by western blot. Long‐term treatment with aripiprazole or N‐acetylcysteine reversed social and cognitive deficits and reduced the exploratory behaviors. In the HIP of 16‐day‐old model pups, H2S levels and MST protein expression were significantly decreased. In adult model rats, H2S levels remained unchanged, bound sulfane sulfur significantly increased, and the expression of CBS and MST slightly decreased. The studied drugs significantly reduced the level of bound sulfane sulfur and the expression of tested enzymes. The reduction in bound sulfane sulfur level coincided with the attenuation of exploratory behavior, suggesting that modulation of anaerobic cysteine metabolism in the HIP may have therapeutic potential in schizophrenia.
... The high proliferation of cancer cells creates redox stress conditions inside the cells, which activates the CBS gene to produce H2S through the 272 CXXC 275 motif [200]. Although some studies showed that CBS expression is downregulated in glioma tumor cells, gastrointestinal cancer cells [186,187,201], and hepatocellular carcinoma, alternatively, reduced CBS expression upregulates the 3-MST gene in glioma tumor cells [202]. ...
... The high proliferation of cancer cells creates redox stress conditions inside the cells, which activates the CBS gene to produce H 2 S through the 272 CXXC 275 motif [200]. Although some studies showed that CBS expression is downregulated in glioma tumor cells, gastrointestinal cancer cells [186,187,201], and hepatocellular carcinoma, alternatively, reduced CBS expression upregulates the 3-MST gene in glioma tumor cells [202]. ...
A high level of homocysteine (Hcy) is associated with oxidative/ER stress, apoptosis, and impairment of angiogenesis, whereas hydrogen sulfide (H2S) has been found to reverse this condition. Recent studies have shown that cancer cells need to produce a high level of endogenous H2S to maintain cell proliferation, growth, viability, and migration. However, any novel mechanism that targets this balance of Hcy and H2S production has yet to be discovered or exploited. Cells require homocysteine metabolism via the methionine cycle for nucleotide synthesis, methylation, and reductive metabolism, and this pathway supports the high proliferative rate of cancer cells. Although the methionine cycle favors cancer cells for their survival and growth, this metabolism produces a massive amount of toxic Hcy that somehow cancer cells handle very well. Recently, research showed specific pathways important for balancing the antioxidative defense through H2S production in cancer cells. This review discusses the relationship between Hcy metabolism and the antiapoptotic, antioxidative, anti-inflammatory, and angiogenic effects of H2S in different cancer types. It also summarizes the historical understanding of targeting antioxidative defense systems, angiogenesis, and other protective mechanisms of cancer cells and the role of H2S production in the genesis, progression, and metastasis of cancer. This review defines a nexus of diet and precision medicine in targeting the delicate antioxidative system of cancer and explores possible future therapeutics that could exploit the Hcy and H2S balance.
... Studies from Diwakar in 2007 have demonstrated that even though the activity of the CSE in the mouse brain represents only 1 % of the liver, CSE inhibition by oxidative stress alters GSH levels since CSE is a rate-limiting enzyme for Cys synthesis from cystathionine; thus CSE activity maintains GSH homeostasis in the brain and preserves mitochondrial function, which is altered in PD patients (Diwakar and Ravindranath 2007). In addition, H 2 S is also produced in the brain by the activity of 3-MPST (Kimura et al. 2015). Shibuya et al. (2009) have demonstrated that brain homogenates of CBS-knockout mice can produce H 2 S via 3-MPST enzyme in similar levels as wildtype mice (Shibuya et al. 2009). ...
The transsulfuration pathway (TSP) is a metabolic pathway involving sulfur transfer from homocysteine to cysteine. Transsulfuration pathway leads to many sulfur metabolites, principally glutathione, H2S, taurine, and cysteine. Key enzymes of the TSP, such as cystathionine β-synthase and cystathionine γ-lyase, are essential regulators at multiple levels in this pathway. TSP metabolites are implicated in many physiological processes in the central nervous system and other tissues. TSP is important in controlling sulfur balance and optimal cellular functions such as glutathione synthesis. Alterations in the TSP and related pathways (transmethylation and remethylation) are altered in several neurodegenerative diseases, including Parkinson's disease, suggesting their participation in the pathophysiology and progression of these diseases. In Parkinson's disease many cellular processes are comprised mainly those that regulate redox homeostasis, inflammation, reticulum endoplasmic stress, mitochondrial function, oxidative stress, and sulfur content metabolites of TSP are involved in these damage processes. Current research on the transsulfuration pathway in Parkinson's disease has primarily focused on the synthesis and function of certain metabolites, particularly glutathione. However, our understanding of the regulation of other metabolites of the transsulfuration pathway, as well as their relationships with other metabolites, and their synthesis regulation in Parkinson´s disease remain limited. Thus, this paper highlights the importance of studying the molecular dynamics in different metabolites and enzymes that affect the transsulfuration in Parkinson's disease.
... Subsequent studies using various model systems further established the supporting role of H 2 S in cognitive functions mediated by NMDA receptors, which included increased sulfhydration of NMDA receptor subunits (Li et al., 2017;Tu et al., 2016;Xu et al., 2015). These reports prompted detailed studies of the localization of H 2 S-generating enzymes in the brain, and it is now wellestablished that CSE is localized to neurons, CBS is preferentially expressed in Bergman glia and astrocytes (Enokido et al., 2005;Linden et al., 2008;Morikawa et al., 2012), and MPST is present in both neurons and astrocytes (Kimura et al., 2015;Shibuya et al., 2009). ...
The gaseous neurotransmitter hydrogen sulfide (H2 S) exerts neuroprotective efficacy in the brain via post-translational modification of cysteine residues by sulfhydration, also known as persulfidation. This process is comparable in biological impact to phosphorylation and mediates a variety of signaling events. Unlike conventional neurotransmitters, H2 S cannot be stored in vesicles due to its gaseous nature. Instead, it is either locally synthesized or released from endogenous stores. Sulfhydration affords both specific and general neuroprotective effects and is critically diminished in several neurodegenerative disorders. Conversely, some forms of neurodegenerative disease are linked to excessive cellular H2 S. Here, we review the signaling roles of H2 S across the spectrum of neurodegenerative diseases, including Huntington's disease, Parkinson's disease, Alzheimer's disease, Down syndrome, traumatic brain injury, the ataxias, and amyotrophic lateral sclerosis, as well as neurodegeneration generally associated with aging.
... For H 2 S n and elemental sulfur (S 8 ) analysis, the sample was derivatized with methyl trifluoromethanesulfonate in methanol to convert H 2 S n to dimethylpolysulfide and solubilize S 8 ; both dimethylpolysulfide and S 8 can be detected using HPLC with UV detection (33,34). For sulfide, GSH, and GSSH analysis, the sample was centrifuged at 13,000 Â g for 2 min, and 50 ml of the supernatant was derivatized with mBBr and detected using HPLC, according to a published method (65). For GS n H, GS n G, and H 2 S n analysis, the supernatant was derivatized with HPE-IAM and detected using LC-MS, according to a published method (31). ...
Sulfur-oxidizing bacteria can oxidize hydrogen sulfide (H 2 S) to produce sulfur globules. Although the process is common, the pathway is unclear. In recombi-nant Escherichia coli and wild-type Corynebacterium vitaeruminis DSM 20294 with sul-fide:quinone oxidoreductase (SQR) but no enzymes to oxidize zero valence sulfur, SQR oxidized H 2 S into short-chain inorganic polysulfide (H 2 S n , n $ 2) and organic polysul-fide (RS n H, n $ 2), which reacted with each other to form long-chain GS n H (n $ 2) and H 2 S n before producing octasulfur (S 8), the main component of elemental sulfur. GS n H also reacted with glutathione (GSH) to form GS n G (n $ 2) and H 2 S; H 2 S was again oxidized by SQR. After GSH was depleted, SQR simply oxidized H 2 S to H 2 S n , which spontaneously generated S 8. S 8 aggregated into sulfur globules in the cyto-plasm. The results highlight the process of sulfide oxidation to S 8 globules in the bacterial cytoplasm and demonstrate the potential of using heterotrophic bacteria with SQR to convert toxic H 2 S into relatively benign S 8 globules. IMPORTANCE Our results provide evidence of H 2 S oxidation producing octasulfur globules via sulfide:quinone oxidoreductase (SQR) catalysis and spontaneous reactions in the bacterial cytoplasm. Since the process is an important event in geo-chemical cycling, a better understanding facilitates further studies and provides theoretical support for using heterotrophic bacteria with SQR to oxidize toxic H 2 S into sulfur globules for recovery.
... Disulfane (HSSH), trisulfane, (HSSSH) and their conjugate bases hydrodisulfide or disulfanide (HS 2 ‾) and hydrotrisulfide (HS 3 ‾), respectively, are the lower members of the family of polysulfanes (HSS n H, 1 ≤ n ≤ 34) [1][2][3], and have been recognized as endogenous diffusible species in different tissues [4][5][6][7][8][9]. These inorganic reactive sulfur species (RSS) appear as byproducts of the activity of the 3-mercaptopyruvate sulfuryl transferase, 3-MST, which canonically delivers hydrogen sulfide, H 2 S [10,11]. The regulated formation of HSS n H, and their conjugate bases polysulfides, HSS n ‾, by the 3-MST remains unknown [12,13]. ...
The mechanism of the metal centered reduction of metmyoglobin (MbFeIII) by inorganic disulfide species has been studied by combined spectroscopic and kinetic analyses, under argon atmosphere. The process is kinetically characterized by biexponential time traces, for variable ratios of excess disulfide to protein, in the pH interval 6.6-8.0. Using UV-vis and resonance Raman spectroscopies, we observed that MbFeIII is converted into a low spin hexacoordinated ferric complex, tentatively assigned as MbFeIII(HSS-)/MbFeIII(SS2-), in an initial fast step. The complex is slowly converted into a pentacoordinated ferrous form, assigned as MbFeII according to the resonance Raman records. The reduction is a pH-dependent process, but independent of the initial disulfide concentration, suggesting the unimolecular decomposition of the intermediate complex following a reductive homolysis. We estimated the rate of the fast formation of the complex at pH 7.4 (kon = 3.7 × 103 M-1 s-1), and a pKa2 = 7.5 for the equilibrium MbFeIII(HSS-)/MbFeIII(SS2-). Also, we estimated the rate for the slow reduction at the same pH (kred = 10-2 s-1). A reaction mechanism compliant with the experimental results is proposed. This mechanistic study provides a differential kinetic signature for the reactions of disulfide compared to sulfide species on metmyoglobin, which may be considered in other hemeprotein systems.
... However, it has been observed previously that overexpression of MPST increases intracellular 'bound' sulfane sulfur (S 0 ) content, potentially indicating a direct role for MPST in general protein persulfidation [20][21][22][23] . Nonetheless, this possibility has not been tested so far. ...
... While the above-described experiments indicated direct proteinto-protein transpersulfidation, they did not rule out an alternative possibility. Previous studies have suggested that MPST can generate inorganic polysulfides (H 2 S 2 , H 2 S 3 ) 22,23 , which in principle could facilitate target protein persulfidation (for example, P-SH + H 2 S 2 → P-SSH + H 2 S). ...
... Previously, it has been suggested that MPST is capable of generating inorganic polysulfides, including H 2 S 2 (ref. 22 ). We therefore asked whether the observed sulfur transfer from MPST to roGFP2 is indeed a direct one or whether it may be mediated through a diffusible LMW inorganic polysulfide. ...
Protein S-persulfidation (P-SSH) is recognized as a common posttranslational modification. It occurs under basal conditions and is often observed to be elevated under stress conditions. However, the mechanism(s) by which proteins are persulfidated inside cells have remained unclear. Here we report that 3-mercaptopyruvate sulfur transferase (MPST) engages in direct protein-to-protein transpersulfidation reactions beyond its previously known protein substrates thioredoxin and MOCS3/Uba4, associated with H2S generation and transfer RNA thiolation, respectively. We observe that depletion of MPST in human cells lowers overall intracellular protein persulfidation levels and identify a subset of proteins whose persulfidation depends on MPST. The predicted involvement of these proteins in the adaptation to stress responses supports the notion that MPST-dependent protein persulfidation promotes cytoprotective functions. The observation of MPST-independent protein persulfidation suggests that other protein persulfidases remain to be identified.
... In contrast, CysSSH may also be formed by the reaction between CysSH and bound sulfane sulfur species (BSS) [66]. BSS, which encompass sulfur-derivatives with a sulfur formal oxidation state of -I and 0 [66], are notably produced in cells by hydrogen sulfide [67] or 3-mercaptopyruvic acid [68] metabolism. Although their exact speciation is challenging to establish, their global concentrations (from high nM to 50-100 µM, depending on the studies) [49,66] are orders of magnitude higher than those detected for hydrogen sulfide or H2O2 in various biological media under physiological conditions [66]. ...
DJ-1 (also called PARK7) is a ubiquitously expressed protein involved in the etiology of Parkinson disease and cancers. At least one of its three cysteine residues is functionally essential, and its oxidation state determines the specific function of the enzyme. DJ-1 was recently reported to be persulfidated in mammalian cell lines, but the implications of this post-translational modification have not yet been analyzed. Here, we report that recombinant DJ-1 is reversibly persulfidated at cysteine 106 by reaction with various sulfane donors and subsequently inhibited. Strikingly, this reaction is orders of magnitude faster than C106 oxidation by H2O2, and persulfidated DJ-1 behaves differently than sulfinylated DJ-1. Both these PTMs most likely play a dedicated role in DJ-1 signaling or protective pathways.
