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Nature Communications Nov 2023Sulfonation as one of the most important modification reactions in nature is essential for many biological macromolecules to function. Development of green sulfonate...
Sulfonation as one of the most important modification reactions in nature is essential for many biological macromolecules to function. Development of green sulfonate group donor regeneration systems to efficiently sulfonate compounds of interest is always attractive. Here, we design and engineer two different sulfonate group donor regeneration systems to boost the biosynthesis of sulfated compounds. First, we assemble three modules to construct a 3'-phosphoadenosine-5'-phosphosulfate (PAPS) regeneration system and demonstrate its applicability for living cells. After discovering adenosine 5'-phosphosulfate (APS) as another active sulfonate group donor, we engineer a more simplified APS regeneration system that couples specific sulfotransferase. Next, we develop a rapid indicating system for characterizing the activity of APS-mediated sulfotransferase to rapidly screen sulfotransferase variants with increased activity towards APS. Eventually, the active sulfonate group equivalent values of the APS regeneration systems towards trehalose and p-coumaric acid reach 3.26 and 4.03, respectively. The present PAPS and APS regeneration systems are environmentally friendly and applicable for scaling up the biomanufacturing of sulfated products.
Topics: Sulfates; Phosphoadenosine Phosphosulfate; Sulfotransferases; Kinetics
PubMed: 37949843
DOI: 10.1038/s41467-023-43195-1 -
Journal of Crohn's & Colitis Mar 2024Carbohydrate sulfotransferase 15 [CHST15] biosynthesizes sulphated matrix glycosaminoglycans and is implicated in intestinal inflammation and fibrosis. Here, we evaluate... (Randomized Controlled Trial)
Randomized Controlled Trial
BACKGROUND AND AIMS
Carbohydrate sulfotransferase 15 [CHST15] biosynthesizes sulphated matrix glycosaminoglycans and is implicated in intestinal inflammation and fibrosis. Here, we evaluate the efficacy and safety of the double-stranded RNA oligonucleotide GUT-1, a specific blocker of CHST15, as induction therapy in patients with ulcerative colitis [UC].
METHODS
In this randomized, double-blind, placebo-controlled, phase 2a study, we enrolled endoscopically active UC patients, refractory to conventional therapy, in five hospital centres across Germany. Patients were randomized 1:1:1 using a block randomized technique to receive a single dosing of 25 nM GUT-1, 250 nM GUT-1, or placebo by endoscopic submucosal injections. The primary outcome measure was improvement of endoscopic lesions at weeks 2 or 4. The secondary outcome measures included clinical and histological responses. Safety was assessed in all patients who received treatment.
RESULTS
Twenty-eight patients were screened, 24 were randomized, and 21 were evaluated. Endoscopic improvement at weeks 2 or 4 was achieved by 71.4% in the GUT-1 250 nM, 0% in the GUT-1 25 nM, and 28.6% in the placebo group. Clinical remission was shown by 57.1% in the GUT-1 250 nM, 0% in the GUT-1 25 nM, and 14.3% in the placebo groups. Histological improvement was shown by 42.9% in the GUT-1 250 nM, 0% in the GUT-1 25 nM, and 0% in the placebo groups. GUT-1 250 nM reduced CHST15 expression significantly and suppressed mucosal inflammation and fibrosis. GUT-1 application was well tolerated.
CONCLUSION
Single dosing by submucosal injection of GUT-1 repressed CHST15 mucosal expression and may represent a novel induction therapy by modulating tissue remodelling in UC.
Topics: Humans; Colitis, Ulcerative; RNA; Oligonucleotides; Fibrosis; Inflammation
PubMed: 37777210
DOI: 10.1093/ecco-jcc/jjad162 -
Plant Physiology and Biochemistry : PPB Oct 2023The present study investigated the effects regulating melatonin (MT) biosynthesis under methyl jasmonate (MeJA) treatment in mustard sprouts. The results revealed that...
The present study investigated the effects regulating melatonin (MT) biosynthesis under methyl jasmonate (MeJA) treatment in mustard sprouts. The results revealed that MeJA significantly increased the MT content in the sprouts to 11.43 times that of the control. However, MeJA treatment had an inhibitory effect on growth. Tryptophan decarboxylase and tryptamine 5-hydroxylase gene expression were significantly induced by MeJA. Moreover, 156 differential abundance proteins (DAPs) were detected in 4-day-old sprouts using quantitative proteomic methods. These DAPs were divided into 13 functional groups, and the vast majority of DAPs involved in defense/stress, energy, signal transduction, and secondary metabolism increased. MeJA treatment significantly enriched 15 pathways, including glutathione metabolism, biosynthesis of secondary metabolites, and tryptophan metabolism. In particular, the abundance of three DAPs (myrosinase 1, cytosolic sulfotransferase 16, and glutamate-glyoxylate aminotransferase 2) in the tryptophan metabolism pathway, a substrate for MT biosynthesis, increased significantly. In summary, MeJA induces endogenous MT biosynthesis in mustard sprouts by promoting the genes expression of MT synthetase and increasing the abundance of tryptophan-related proteins.
PubMed: 37751654
DOI: 10.1016/j.plaphy.2023.108055 -
Nature Communications Feb 2024Heparan sulfate (HS) polysaccharides are major constituents of the extracellular matrix, which are involved in myriad structural and signaling processes. Mature HS...
Heparan sulfate (HS) polysaccharides are major constituents of the extracellular matrix, which are involved in myriad structural and signaling processes. Mature HS polysaccharides contain complex, non-templated patterns of sulfation and epimerization, which mediate interactions with diverse protein partners. Complex HS modifications form around initial clusters of glucosamine-N-sulfate (GlcNS) on nascent polysaccharide chains, but the mechanistic basis underpinning incorporation of GlcNS itself into HS remains unclear. Here, we determine cryo-electron microscopy structures of human N-deacetylase-N-sulfotransferase (NDST)1, the bifunctional enzyme primarily responsible for initial GlcNS modification of HS. Our structures reveal the architecture of both NDST1 deacetylase and sulfotransferase catalytic domains, alongside a non-catalytic N-terminal domain. The two catalytic domains of NDST1 adopt a distinct back-to-back topology that limits direct cooperativity. Binding analyses, aided by activity-modulating nanobodies, suggest that anchoring of the substrate at the sulfotransferase domain initiates the NDST1 catalytic cycle, providing a plausible mechanism for cooperativity despite spatial domain separation. Our data shed light on key determinants of NDST1 activity, and describe tools to probe NDST1 function in vitro and in vivo.
Topics: Humans; Cryoelectron Microscopy; Heparitin Sulfate; Catalytic Domain; Sulfotransferases; Extracellular Matrix
PubMed: 38351061
DOI: 10.1038/s41467-024-45419-4 -
Acta Pharmaceutica Sinica. B Jun 2024Although sulfonation plays crucial roles in various biological processes and is frequently utilized in medicinal chemistry to improve water solubility and chemical...
Although sulfonation plays crucial roles in various biological processes and is frequently utilized in medicinal chemistry to improve water solubility and chemical diversity of drug leads, it is rare and underexplored in ribosomally synthesized and post-translationally modified peptides (RiPPs). Biosynthesis of RiPPs typically entails modification of hydrophilic residues, which substantially increases their chemical stability and bioactivity, albeit at the expense of reducing water solubility. To explore sulfonated RiPPs that may have improved solubility, we conducted co-occurrence analysis of RiPP class-defining enzymes and sulfotransferase (ST), and discovered two distinctive biosynthetic gene clusters (BGCs) encoding both lanthipeptide synthetase (LanM) and ST. Upon expressing these BGCs, we characterized the structures of novel sulfonated lanthipeptides and determined the catalytic details of LanM and ST. We demonstrate that SslST-catalyzed sulfonation is leader-independent but relies on the presence of A ring formed by LanM. Both LanM and ST are promiscuous towards residues in the A ring, but ST displays strict regioselectivity toward Tyr5. The recognition of cyclic peptide by ST was further discussed. Bioactivity evaluation underscores the significance of the ST-catalyzed sulfonation. This study sets up the starting point to engineering the novel lanthipeptide STs as biocatalysts for hydrophobic lanthipeptides improvement.
PubMed: 38828142
DOI: 10.1016/j.apsb.2024.02.016 -
The Science of the Total Environment Sep 2023As the herbicide acetochlor (ACT) affects the plankton food web, this study investigated the effects of ACT and exocrine infochemicals from daphnids (after ACT exposure...
As the herbicide acetochlor (ACT) affects the plankton food web, this study investigated the effects of ACT and exocrine infochemicals from daphnids (after ACT exposure and/or starvation) on Scenedesmus obliquus growth, as well as the effects of ACT and starvation on life-history traits of Daphnia magna. Filtered secretions from daphnids increased algal ACT tolerance, dependent on different ACT exposure histories and food intake. Endogenous and secretory metabolite profiles of daphnids following ACT and/or starvation appear to be regulated by the fatty acid synthesis pathway and sulfotransferases and were related to energy allocation trade-offs. Oleic acid (OA) and octyl sulfate (OS), screened by secreted and somatic metabolomics, affected algal growth and ACT behavior in the algal culture in opposite ways. Trophic and non-trophic interspecific effects caused by ACT were observed in microalgae-daphnid microcosms, including algal growth inhibition, daphnid starvation, OA down-regulation, and OS up-regulation. Based on these findings, risk assessment of ACT on the freshwater plankton communities should take species interactions into account.
Topics: Animals; Microalgae; Food Chain; Nutritional Status; Daphnia
PubMed: 37301391
DOI: 10.1016/j.scitotenv.2023.164718 -
Molecules (Basel, Switzerland) Oct 2023The aim of this study was to develop and optimize a chiral HPLC-MS/MS method for quantitative analysis of ()-/()-salbutamol and ()-/()-salbutamol-4'--sulfate in human...
Development of an HPLC-MS/MS Method for Chiral Separation and Quantitation of ()- and ()-Salbutamol and Their Sulfoconjugated Metabolites in Urine to Investigate Stereoselective Sulfonation.
The aim of this study was to develop and optimize a chiral HPLC-MS/MS method for quantitative analysis of ()-/()-salbutamol and ()-/()-salbutamol-4'--sulfate in human urine to allow for bioanalytical quantitation of the targeted analytes and investigations of stereoselectivity in the sulfonation pathway of human phase Ⅱ metabolism. For analytical method development, a systematic screening of columns and mobile phases to develop a separation via enantiomerically selective high performance liquid chromatography was performed. Electrospray ionization settings were optimized via multiple-step screening and a full factorial design-of-experiment. Both approaches were performed matrix-assisted and the predicted values were compared. The full factorial design was superior in terms of prediction power and knowledge generation. Performing a longitudinal excretion study in one healthy volunteer allowed for the calculation of excretion rates for all four targeted analytes. For this proof-of-concept, either racemic salbutamol or enantiopure levosalbutamol was administered perorally or via inhalation, respectively. A strong preference for sulfonation of ()-salbutamol for inhalation and peroral application was found in experiments. In previous studies phenol sulfotransferase 1A3 was described to be mainly responsible for salbutamol sulfonation in humans. Thus, in vitro and in silico investigations of the stereoselectivity of sulfotransferase 1A3 complemented the study and confirmed these findings.
Topics: Humans; Albuterol; Chromatography, High Pressure Liquid; Tandem Mass Spectrometry; Levalbuterol; Administration, Inhalation; Stereoisomerism
PubMed: 37894685
DOI: 10.3390/molecules28207206 -
Apoptosis : An International Journal on... Jun 2024The cytosolic sulfotransferases (SULTs) are phase II conjugating enzymes, which are widely expressed in the liver and mainly mediate the sulfation of numerous...
The cytosolic sulfotransferases (SULTs) are phase II conjugating enzymes, which are widely expressed in the liver and mainly mediate the sulfation of numerous xenobiotics and endogenous compounds. However, the role of various SULTs genes has not been reported in hepatocellular carcinoma (HCC). This study aims to analyze the expression and potential functional roles of SULTs genes in HCC and to identify the role of SULT2A1 in HCC stemness as well as the possible mechanism. We found that all of the 12 SULTs genes were differentially expressed in HCC. Moreover, clinicopathological features and survival rates were also investigated. Multivariate regression analysis showed that SULT2A1 and SULT1C2 could be used as independent prognostic factors in HCC. SULT1C4, SULT1E1, and SULT2A1 were significantly associated with immune infiltration. SULT2A1 deficiency in HCC promoted chemotherapy resistance and stemness maintenance. Mechanistically, silencing of SULT2A1 activated the AKT signaling pathway, on the one hand, promoted the expression of downstream stemness gene c-Myc, on the other hand, facilitated the NRF2 expression to reduce the accumulation of ROS, and jointly increased HCC stemness. Moreover, knockdown NR1I3 was involved in the transcriptional regulation of SULT2A1 in stemness maintenance. In addition, SULT2A1 knockdown HCC cells promoted the proliferation and activation of hepatic stellate cells (HSCs), thereby exerting a potential stroma remodeling effect. Our study revealed the expression and role of SULTs genes in HCC and identified the contribution of SULT2A1 to the initiation and progression of HCC.
Topics: Carcinoma, Hepatocellular; Liver Neoplasms; Sulfotransferases; Gene Knockdown Techniques; Humans; Animals; Mice; Mice, Inbred BALB C; Mutation; Gene Expression Regulation, Neoplastic; DNA Methylation; Drug Resistance, Neoplasm; Embryonic Stem Cells; Prognosis; Cell Line, Tumor
PubMed: 38411862
DOI: 10.1007/s10495-024-01938-5 -
Journal of Integrative Plant Biology Aug 2023Phytohormone abscisic acid (ABA) plays vital roles in stress tolerance, while long-term overactivation of ABA signaling suppresses plant growth and development. However,...
Phytohormone abscisic acid (ABA) plays vital roles in stress tolerance, while long-term overactivation of ABA signaling suppresses plant growth and development. However, the braking mechanism of ABA responses is not clear. Protein tyrosine sulfation catalyzed by tyrosylprotein sulfotransferase (TPST) is a critical post-translational modification. Through genetic screening, we identified a tpst mutant in Arabidopsis that was hypersensitive to ABA. In-depth analysis revealed that TPST could interact with and sulfate SnRK2.2/2.3/2.6, which accelerated their degradation and weakened the ABA signaling. Taken together, these findings uncovered a novel mechanism of desensitizing ABA responses via protein sulfation.
Topics: Arabidopsis; Plant Growth Regulators; Abscisic Acid; Sulfotransferases; Arabidopsis Proteins; Protein Serine-Threonine Kinases
PubMed: 37052306
DOI: 10.1111/jipb.13493 -
Chemical Biology & Drug Design Nov 2023Human sulfotransferases 1A3 (SULT1A3) has received particular interest, due to their functions of catalyzing the sulfonation of numerous phenolic substrates, including...
Human sulfotransferases 1A3 (SULT1A3) has received particular interest, due to their functions of catalyzing the sulfonation of numerous phenolic substrates, including bioactive endogenous molecules and therapeutic agents. However, the regulation of SULT1A3 expression and the underlying mechanism remain unclear. Here, we aimed to investigate the regulation effects of bile acid-activated farnesoid X receptor (FXR) on SULT1A3 expression, and to shed light on the mechanism thereof. Our results demonstrated that FXR agonists (CDCA and GW4064) significantly inhibit the expression of SULT1A3 at mRNA and protein levels. In addition, overexpression of FXR led to decrease in SULT1A3 expression and knockdown of FXR significantly induced the expression of SULT1A3 in protein and mRNA levels, confirming that FXR expression manifestly showed negative regulatory effect on basal SULT1A3 expression. Furthermore, a combination of luciferase reporter gene and CHIP assays showed that FXR repressed SULT1A3 transcription through direct binding to the region at base pair positions -664 to -654. In conclusion, this study for the first time confirmed FXR was a negative transcriptional regulator of human SULT1A3 enzyme.
Topics: Humans; Chenodeoxycholic Acid; Promoter Regions, Genetic; Receptors, Cytoplasmic and Nuclear; RNA, Messenger; Sulfotransferases
PubMed: 37487659
DOI: 10.1111/cbdd.14312