Improved tag-switch method reveals that thioredoxin acts as depersulfidase and controls the intracellular levels of protein persulfidation

Wedmann, Rudolf; Onderka, Constantin; Wei, Shengwei; Szijártó, István András; Miljković, Jan Lj.; Mitrović, Aleksandra; Lange, M.; Savitsky, Sergey; Yadav, Pramod Kumar; Torregrossa, Roberta; Harrer, E.; Harrer, Thomas; Ishii, Isao; Gollasch, Maik; Wood, Mark E.; Galardon, Erwan; Xian, Ming; Whiteman, Matthew; Banerjee, Ruma; Filipović, Miloš R.

doi:10.1039/c5sc04818d

Cited by 180 publications

(195 citation statements)

References 48 publications

(67 reference statements)

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“…Thioredoxin is postulated to play a role in H 2 S signaling and inhibition of thioredoxin reductase is associated with increased intracellular persulfide levels (46,47). The concentration of thioredoxin is reportedly ~8-10 µM in liver (48) (Fig.…”

Section: Discussionmentioning

confidence: 99%

“…For instance in rhodanese, which can catalyze many of the same reactions as TSTD1, the deeply set active site pocket likely promotes interactions with low molecular weight substrates like GSSH whereas in TSTD1, the shallow active site pocket might promote interactions with larger substrates, namely proteins. In addition, the sequestered active sites in rhodanese and MST likely contribute to stabilizing the Cys-SSH intermediate, which has been seen in their crystal structures (13,45).Thioredoxin is postulated to play a role in H 2 S signaling and inhibition of thioredoxin reductase is associated with increased intracellular persulfide levels (46,47). The concentration of thioredoxin is reportedly ~8-10 µM in liver (48) (Fig.…”

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confidence: 96%

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Thiosulfate sulfurtransferase-like domain–containing 1 protein interacts with thioredoxin

Libiad

Motl

Akey

et al. 2018

Journal of Biological Chemistry

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ǂ These authors contributed equally to this work Rhodanese domains are structural modules present in the sulfurtransferase superfamily. These domains can exist as single units, in tandem repeats or fused to domains with other activities. Despite their prevalence across species, the specific physiological roles of most sulfurtransferases are not known. Mammalian rhodanese and mercaptopyruvate sulfurtransferase are perhaps the best-studied members of this protein superfamily and are involved in hydrogen sulfide metabolism. The relatively unstudied human thiosulfate sulfurtransferase like domain-containing 1 (TSTD1) protein, a single-domain cytoplasmic sulfurtransferase, was also postulated to play a role in the sulfide oxidation pathway using thiosulfate to form glutathione persulfide, for subsequent processing in the mitochondrial matrix. Prior kinetic analysis of TSTD1 was performed at pH 9.2, raising questions about relevance and the proposed model for TSTD1 function. In this study, we report a 1.04 Å resolution crystal structure of human TSTD1, which displays an exposed active site that is distinct from that of rhodanese and mercaptopyruvate sulfurtransferase. Kinetic studies with a combination of sulfur donors and acceptors reveal that TSTD1 exhibits a low K M for thioredoxin as a sulfane sulfur acceptor and that it utilizes thiosulfate inefficiently as a sulfur donor. The active site exposure and its interaction with thioredoxin, suggest that TSTD1 might play a role in sulfide-based signaling. The apical localization of TSTD1 in human colonic crypts, which interfaces with sulfide-releasing microbes, and the overexpression of TSTD1 in colon cancer, provides potentially intriguing clues as to its role in sulfide metabolism. Hydrogen sulfide (H 2 S)1 is an important signaling molecule with effects on multiple physiological processes including neuromodulation, inflammation and cardiac function (1-7). Maintaining healthy levels of H 2 S in mammalian cells requires tight control of its biosynthesis and its catabolism (8,9). H 2 S is produced endogenously by two enzymes in the transsulfuration pathway, cystathionine β-synthase and γ-cystathionase as well as by mercaptopyruvate sulfurtransferase (MST) (10-13). H 2 S is oxidized via the sulfide oxidation pathway to generate the end-products thiosulfate Protein persulfidation, a posttranslational modification of cysteine residues, is postulated to be a major mechanism by which H 2 S signals (17). However, the mechanism by which target proteins acquire this modification and the sulfur source(s) used in persulfidation reactions are unclear (18). Sulfurtransferases like rhodanese or the single rhodanese domain containing TSTD1 could potentially play a role in protein persulfidation because of their capacity to form an active site cysteine persulfide during their reaction cycle (18).The rhodanese superfamily members are distinguished by a structural module with an α/β topology in which α-helices surround a central five-stranded β-sheet core (19). At least three variations...

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Section: Discussionmentioning

confidence: 99%

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confidence: 96%

Thiosulfate sulfurtransferase-like domain–containing 1 protein interacts with thioredoxin

Libiad

Motl

Akey

et al. 2018

Journal of Biological Chemistry

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“…Emerging evidence suggests that highly oxidized sulfur species, termed reactive sulfur species (RSS), are the actual sulfide-signaling species (13-16). These include HS•, cysteine sulfenic acids, cysteine persulfides, glutathione persulfides (GSSHs), and inorganic polysulfide species, which catalyze persulfidation of specific proteins and enzymes (13,(17)(18)(19)(20), although the functional impact of this protein modification is as yet not generally clear (17).Hydrogen sulfide inhibits respiration by poisoning cytochrome oxidase and binding and precipitating biologically required divalent transition metals, and is thus toxic at high concentrations (21). The ability to adapt to a sulfide-rich environment is therefore critical for the survival for some microorganisms.…”

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confidence: 99%

Sulfide-responsive transcriptional repressor SqrR functions as a master regulator of sulfide-dependent photosynthesis

Shimizu

Shen

Fang

et al. 2017

Proc. Natl. Acad. Sci. U.S.A.

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Sulfide was used as an electron donor early in the evolution of photosynthesis, with many extant photosynthetic bacteria still capable of using sulfur compounds such as hydrogen sulfide (H 2 S) as a photosynthetic electron donor. Although enzymes involved in H 2 S oxidation have been characterized, mechanisms of regulation of sulfide-dependent photosynthesis have not been elucidated. In this study, we have identified a sulfide-responsive transcriptional repressor, SqrR, that functions as a master regulator of sulfide-dependent gene expression in the purple photosynthetic bacterium Rhodobacter capsulatus. SqrR has three cysteine residues, two of which, C41 and C107, are conserved in SqrR homologs from other bacteria. Analysis with liquid chromatography coupled with an electrospray-interface tandem-mass spectrometer reveals that SqrR forms an intramolecular tetrasulfide bond between C41 and C107 when incubated with the sulfur donor glutathione persulfide. SqrR is oxidized in sulfidestressed cells, and tetrasulfide-cross-linked SqrR binds more weakly to a target promoter relative to unmodified SqrR. C41S and C107S R. capsulatus SqrRs lack the ability to respond to sulfide, and constitutively repress target gene expression in cells. These results establish that SqrR is a sensor of H 2 S-derived reactive sulfur species that maintain sulfide homeostasis in this photosynthetic bacterium and reveal the mechanism of sulfide-dependent transcriptional derepression of genes involved in sulfide metabolism.sulfide sensor | photosynthesis regulation | reactive sulfur species | purple bacteria | Rhodobacter T he discovery of ∼550 deep-sea hydrothermal vents more than 30 y ago (1) has led to the theory that energy metabolism in early ancestral organisms may have arisen from deep-sea hydrothermal vents where simple inorganic molecules such as hydrogen sulfide or hydrogen gas, as well as methane, exist (2-4). Such ancient energy metabolism has been assumed to be similar to that of extant chemolithotrophs, which obtain energy from these molecules. Indeed, various chemolithoautotrophic microbes thrive in deep-sea hydrothermal vents and are capable of oxidizing sulfides, methane, and/or hydrogen gas for use as energy sources and electron donors (5). Some photosynthetic bacteria have also been isolated from deep-sea hydrothermal vents that can grow photosynthetically using sulfide as an electron donor and geothermal radiation as an energy source instead of solar radiation (6), as hypothesized for ancestral phototrophs.Many purple photosynthetic bacteria have remarkable metabolic versatility required to meet the energy demands of sulfidedependent and -independent photosynthesis as well as aerobic and anaerobic respiration. These bacteria tightly control the synthesis of their electron transfer proteins involved in each growth mode in response to a specific electron donor, oxygen tension, and light intensity (7, 8). Among these regulatory systems, oxygen-and lightsensing mechanisms have been well-studied; however, mechanisms used to sen...

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“…22 Similar to NO and CO, H2S may directly regulate target proteins by S-sulfhydration to elicit its biological and pharmacological responses. 23,24 In this protein post-translational modification, H2S reacts with a free thiol group in active cysteine residue of target protein to form hydropersulfide group (-SSH). 23 It also has been reported that sulfane sulfur-containing compounds have more reactive activity in mediating protein S-sulfhydration in comparison with H2S.…”

Section: H2s and Protein S-sulfhydrationmentioning

confidence: 99%

“…23 It also has been reported that sulfane sulfur-containing compounds have more reactive activity in mediating protein S-sulfhydration in comparison with H2S. 24,25 The modified biotin switch assay, mleimide assay, and mass spectrometry are often used to detect protein S-sulfhydration. 21 Thus far, a handful of proteins have been demonstrated to be targeted by H2S for S-sulfhydration, including Keap1, GAPDH, NF-κB, MEK1, Parkin, PTP-1B, pyruvate carboxylase, and many others.…”

Section: H2s and Protein S-sulfhydrationmentioning

confidence: 99%

Gasotransmitters and Protein Post-Translational Modifications

Yang¹

2017

MOJPB

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Nitric oxide (NO), carbon monoxide (CO) and hydrogen sulfide (H2S), the three members in gasotransmitter family, have emerged as important regulators of cellular functions and pathophysiological responses. In this mini-review, the current understanding on the roles of these gasotransmitters in regulating cellular events via post-translational modification proteins is summarized. NO chemically reacts with specific cysteine residue(s) in target proteins via S-nitrosylation. CO alters protein conformation and activity by forming carbonylation in unique amino acids. H2S binds with the free thiol group in active cysteine residue of target protein to form hydropersulfide group, termed as S-sulfhydration. The mechanisms for gasotransmitter modification of proteins and their reversible process are also highlighted.

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Improved tag-switch method reveals that thioredoxin acts as depersulfidase and controls the intracellular levels of protein persulfidation

Abstract: H2S signals via protein persulfidation. To be regulatory the modification will have to be reversible. Using a new method for persulfide detection, we discover this missing link and show that thioredoxin system acts as depersulfidase in vivo.

Cited by 180 publications

References 48 publications

Thiosulfate sulfurtransferase-like domain–containing 1 protein interacts with thioredoxin

Thiosulfate sulfurtransferase-like domain–containing 1 protein interacts with thioredoxin

Sulfide-responsive transcriptional repressor SqrR functions as a master regulator of sulfide-dependent photosynthesis

Gasotransmitters and Protein Post-Translational Modifications

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