bims-supasi Biomed News
on Sulfation pathways and signalling
Issue of 2025–02–16
thirteen papers selected by
Jonathan Wolf Mueller, University of Birmingham
  1. Expression of Versican in the Retina and Its Implication in Retinal Disease.
  2. Streamlining Sulfated Oligosaccharide and Glycan Synthesis with Engineered Mutant 6-SulfoGlcNAcases.
  3. Clinical Utility of Serum Dehydroepiandrosterone Sulfate (DHEA-S) and Serum Anti-Mullerian Hormone (AMH) Levels in Infertile Females.
  4. Investigation on the mechanism of anionic polysaccharides affecting the gastrointestinal digestion fate of sea cucumber collagen fibrils.
  5. Sulfated Undaria pinnatifida polysaccharides inhibit kidney stone formation through crystalline modulation and relieving cellular oxidative damage and inflammation.
  6. Intracrine Formation of Steroid Hormones in Breast Cancer, Epidermal Keratinocyte, Dermal Fibroblast, and Adipocyte Cell Lines Measured by LC-MS/MS.
  7. Biallelic SLC13A1 loss-of-function variants result in impaired sulfate transport and skeletal phenotypes, including short stature, scoliosis, and skeletal dysplasia.
  8. From Pre-Foaling to Late Pregnancy: Cortisol, DHEA(S), Progesterone, 17-β-Estradiol, and Allopregnanolone Hair Concentration Profiles in Standardbred Mares.
  9. mRNA Degradation as a Therapeutic Solution for Mucopolysaccharidosis Type IIIC: Use of Antisense Oligonucleotides to Promote Downregulation of Heparan Sulfate Synthesis.
  10. Lower Cortisol and Dehydroepiandrosterone Sulfate and Higher Food Addiction in Childhood Obesity: Associations With Stress and Dietary Parameters.
  11. Blood to serum concentration ratios for ethyl glucuronide and ethyl sulfate after five drinking episodes.
  12. Rational engineering of isoform-specific hSULT1E1 fluorogenic substrates for functional analysis and inhibitor screening.
  13. Non-Targeted Metabolome Analysis with Low-Dose Selenate-Treated Arabidopsis.
  1. Adv Exp Med Biol. 2025 ;1468 423-427
    Expression of Versican in the Retina and Its Implication in Retinal Disease.
    Andrea E Dillinger, Anja K Hoffmann, Ernst R Tamm.
      Chondroitin sulfate proteoglycans are characterized by their sulfated glycosaminoglycan chains covalently attached to the core protein. Versican, encoded by the CSPG2 gene, is a chondroitin sulfate proteoglycan highly expressed in the human and mouse retina. Due to alternative splicing, it exists in four different isoforms: V0, V1, V2 and V3. Interactions with a variety of proteins lead to its main functions, such as influencing cellular adhesion, migration, proliferation and regulating inflammatory processes. Versican expression is high during retinal development and decreases in the mature tissue, remaining mostly in the retinal pigment epithelium, Bruch's membrane and choroid blood vessel walls. Mutations in the CSPG2 gene resulting in an imbalance of Versican splice variants cause erosive vitreoretinopathy and Wagner disease. Its regulatory role in a variety of functions, especially those influencing extracellular matrix composition and inflammation, suggests a crucial role in the pathomechanisms of retinal degenerative disorders and should be the purpose of future research.
    Keywords:  Chondroitin sulfate proteoglycan; Extracellular matrix; Inflammation; Vitreoretinopathy; Wagner disease
    DOI:  https://doi.org/10.1007/978-3-031-76550-6_69
  2. J Am Chem Soc. 2025 Feb 10.
    Streamlining Sulfated Oligosaccharide and Glycan Synthesis with Engineered Mutant 6-SulfoGlcNAcases.
    Rajneesh K Bains, Feng Liu, Seyed A Nasseri, Jacob F Wardman, Stephen G Withers.
      Sulfation is a common, but poorly understood, post-glycosylational modification (PGM) used to modulate biological function. To deepen our understanding of the roles of various sulfated glycoforms and their relevant binding proteins, we must expand our enzymatic toolkit for their synthesis. Here, we bypass the need for both sulfotransferases and glycosyltransferases by engineering a series of mutants of a 6-SulfoGlcNAcase, from Streptococcus pneumoniae, to directly and efficiently synthesize not only the ubiquitous 6S-GlcNAc-β-1,3-Gal linkage prevalent within host glycans, but also the 6S-GlcNAc-β-1,6-GalNAc commonly observed within core-6 O-glycans, and the more exotic 6S-GlcNAc-β-1,4-GalNAc linkage. We further elaborate these into complex sulfated N-glycan and O-glycan structures of biological relevance. By utilizing the cost-effective activated donor pNP-6S-GlcNAc in conjunction with mutant GH185 6-SulfoGlcNAcases we demonstrate a simple yet powerful in vitro method for generating well-defined sulfated oligosaccharides and glycoforms for use in a variety of applications including glycan arrays, glycan remodeling, and specificity studies with carbohydrate binding proteins such as lectins.
    DOI:  https://doi.org/10.1021/jacs.4c14102
  3. J Coll Physicians Surg Pak. 2025 Feb;35(2): 157-161
    Clinical Utility of Serum Dehydroepiandrosterone Sulfate (DHEA-S) and Serum Anti-Mullerian Hormone (AMH) Levels in Infertile Females.
    Rimsha Sehar, Muhammad Younas, Zujaja Hina Haroon, Muhammad Anwar, Muhammad Usman Munir.
       OBJECTIVE: To determine the correlation of serum dehydroepiandrosterone sulphate and anti-mullerian hormone levels with infertility in females.
    STUDY DESIGN: Cross-sectional study. Place and Duration of the Study: Department of Chemical Pathology, the Armed Forces Institute of Pathology, Rawalpindi, Pakistan, from July 2022 to August 2023.
    METHODOLOGY: About 110 infertile females with infertility were enrolled in this study. Serum thyroid-stimulating hormone, luteinising hormone, testosterone, follicle-stimulating hormone (FSH), estradiol, serum Anti-Mullerian Hormone (AMH), and Dehydroepiandrosterone Sulphate (DHEA-S) were measured at 3rd menstruation day. Females with primary ovarian insufficiency, using androgen supplementation, and on hormonal therapy were excluded.
    RESULTS: Among the participants, 63.6% had primary infertility and 36.4% had secondary infertility. Both primary and secondary infertility had a positive correlation with serum AMH and DHEA-S levels (r = 0.685; p <0.001) and (r = 0.807; p <0.001), respectively. After ultrasound, 54 (49.1%) females were normal, 12 (10.9%) had PCOS, 12 (10.9%) had fibroids, 8 (7.3%) had fallopian tube defects, 10 (9.1%) had endometriosis, and 14 (12.7%) had low antral follicular count. The correlation between serum AMH and DHEA-S with different subgroups is as follows: Normal females (r = 0.731; p <0.001), PCOs (r = -0.232; p = 0.468), fibroids (r = 0.941, p <0.001), fallopian tube defects (r =-0.800; p = 0.017), endometriosis (r = -0.684, p = 0.013), and low antral follicular count (r = 0.643, p = 0.0130).
    CONCLUSION: This study demonstrated a positive correlation between serum AMH and DHEA-S levels in infertile females.
    KEY WORDS: Infertile women, Ovarian reserve, Anti-Mullerian Hormone, Dehydroepiandrosterone sulphate.
    DOI:  https://doi.org/10.29271/jcpsp.2025.02.157
  4. Int J Biol Macromol. 2025 Feb 06. pii: S0141-8130(25)01270-X. [Epub ahead of print]304(Pt 1): 140721
    Investigation on the mechanism of anionic polysaccharides affecting the gastrointestinal digestion fate of sea cucumber collagen fibrils.
    Xuanli Sha, Yuhan Zhang, Yaoguang Chang, Yanchao Wang.
      Previous research has demonstrated that fucosylated chondroitin sulfate (fCS) and fucoidan (FUC) enhance the gastrointestinal digestion of sea cucumber collagen fibrils, whereas kappa-carrageenan (K-car) and sodium alginate (SA) exhibit inhibitory effects. The objective of this study was to investigate the mechanisms by which these anionic polysaccharides modulate collagen fibril digestion. Upon the addition of fCS and FUC, the total hydroxyproline content in the insoluble precipitate was significantly reduced by 73.98% and 63.62%, respectively, after 2 h of gastric digestion. All four anionic polysaccharides were found to interact with the soluble digestion products of collagen fibrils, leading to fluorescence quenching. Notably, fCS and FUC displayed significantly stronger interactions with the digestion products compared to K-car and SA, as evidenced by fluorescence spectroscopy. Furthermore, heat treatment resulted in enhanced adsorption of polysaccharides onto the insoluble digestion products of collagen fibrils. Interestingly, the inclusion of polysaccharides, particularly fCS and FUC, substantially increased the turbidity of intestinal digestion products. These interactions were mediated by specific collagen, such as APA22677.1, PIK60696.1, PIK60691.1, PIK60692.1, PIK60693.1, and AYL88761.1 (NCBI). These findings provide crucial insights into how anionic polysaccharides influence the digestive behavior of collagen fibrils, offering potential applications in food science and nutrition.
    Keywords:  Collagen fibril; Digestion; Polysaccharide
    DOI:  https://doi.org/10.1016/j.ijbiomac.2025.140721
  5. Biomater Sci. 2025 Feb 10.
    Sulfated Undaria pinnatifida polysaccharides inhibit kidney stone formation through crystalline modulation and relieving cellular oxidative damage and inflammation.
    Xue-Wu Chen, Jun Long, Quan Zhang, Ling-Hong Huang, Xin-Yuan Sun.
      Background: Calcium oxalate (CaOx) crystal deposition and its resultant cellular oxidative damage and inflammation are important causes of renal stone formation. It is clinically important to conduct research on multifunctional anti-stone drugs targeting these predisposing factors. Methods: We modified natural Undaria pinnatifida polysaccharide (UPP0) by sulfation via the sulfur trioxide-pyridine method, resulting in four sulfated polysaccharides with varying sulfate group (-OSO3-) contents: 1.59% (UPP0), 6.03% (UPP1), 20.83% (UPP2), and 36.39% (UPP3), and compared their differences in the inhibition of crystalline formation, renal injury, and inflammation in the process of renal stone formation at chemical and cellular levels. Results: The UPPS were able to inhibit the nucleation, growth and aggregation of CaOx crystals in vitro. Among them, UPP3 with the maximum sulfate group content showed the greatest crystallization inhibition ability. The nucleation inhibition and aggregation inhibition of UPP3 at a concentration of 0.5 mg mL-1 were as high as 80.21% and 72.34%, respectively. The CaOx crystal size regulated by UPP3 was significantly reduced from 25.9 ± 2.8 μm to 5.9 ± 1.2 μm. Furthermore, UPPS were observed to up-regulate the expression of the antioxidant enzyme superoxide dismutase (SOD) in cells, reduce the levels of ROS and malonaldehyde (MDA), enhance lysosomal integrity, decrease intracellular Ca2+ levels, inhibit the decline in mitochondrial membrane potential, reduce the production of cellular inflammatory factors (TNF-α, MCP-1, IL-18, and IL-1β), and ultimately inhibit cell apoptosis. Conclusion: UPPS combine multiple biological functions of crystallization regulation, antioxidant and anti-inflammatory, and have important potential in the prevention of kidney stones. Sulfation modification can improve the biological activity of UPP0 and provide a reference for screening and optimization methods of stone drugs.
    DOI:  https://doi.org/10.1039/d4bm01362j
  6. Int J Mol Sci. 2025 Jan 30. pii: 1188. [Epub ahead of print]26(3):
    Intracrine Formation of Steroid Hormones in Breast Cancer, Epidermal Keratinocyte, Dermal Fibroblast, and Adipocyte Cell Lines Measured by LC-MS/MS.
    Emre Karakus, Andreas Schmid, Andreas Schäffler, Stefan A Wudy, Joachim Geyer.
      Peripheral tissues such as skin and adipose tissue play a crucial role in the intracrine formation of sex steroid hormones, complementing the endocrine and paracrine systems. These mechanisms involve the conversion of dehydroepiandrosterone (DHEA) and its sulfated form-DHEAS-into potent androgenic and estrogenic hormones. In vitro studies using tissue-specific cell lines are essential for unraveling the complex intracrine synthesis of these hormones. This study examined the formation of DHEA, androstenedione (A4), testosterone (T), dihydrotestosterone (DHT), and estradiol (E2) from DHEAS in four cell lines: MCF-7 breast cancer cells, HaCaT keratinocytes, human dermal fibroblasts (HDF), and 3T3-L1 preadipocytes and mature adipocytes, using liquid chromatography-mass spectrometry (LC-MS/MS). MCF-7 cells converted DHEAS to DHEA, A4, T, E2, and DHT, while HaCaT cells produced all these steroids except DHT. Mature 3T3-L1 adipocytes produced DHEA, A4, T, and DHT. By contrast, HDF and 3T3-L1 preadipocytes converted DHEAS only to DHEA and A4. This study highlights the vital role of peripheral tissues, such as skin and adipose tissue, for the intracrine formation of sex hormones and underlines the crucial role of in vitro cell culture models to analyze such effects. The data shed light on the significant impact of androgen metabolism in skin and adipose tissue, which is of great relevance for aging, wound healing, obesity, and lipid metabolism.
    Keywords:  3T3-L1 adipocytes; HaCaT; LC-MS/MS; MCF-7; human dermal fibroblast; intracrine signaling; sex steroids
    DOI:  https://doi.org/10.3390/ijms26031188
  7. Genet Med Open. 2025 ;3 101958
    Biallelic SLC13A1 loss-of-function variants result in impaired sulfate transport and skeletal phenotypes, including short stature, scoliosis, and skeletal dysplasia.
    Christina G Tise, Katie Ashton, Lachlan de Hayr, Kun-Di Lee, Omkar L Patkar, Emma Krzesinski, Jennifer A Bassetti, Erin M Carter, Cathleen Raggio, Andreas Zankl, Anas M Khanshour, Kristhen N Atala, Jonathan J Rios, Carol A Wise, Ying Zhu, Futao Zhang, Tony Roscioli, Michael Buckley, Robert J Harvey, Paul A Dawson.
       Purpose: Sulfate is vital for many physiological processes, including the structural and functional maintenance of macromolecules and formation of sulfur-containing compounds essential for cartilage and bone development. SLC13A1 is a sodium-sulfate cotransporter primarily expressed in the kidney, where it mediates sulfate reabsorption and maintenance of circulating sulfate levels. In this study, we characterized the clinical, biochemical, and functional impact of biallelic SLC13A1 nonsense and/or missense variants in individuals presenting with a skeletal phenotype.
    Methods: Probands were identified by exome or genome sequencing and GeneMatcher. Sulfate levels were quantified using ion chromatography. SLC13A1 missense variants p.(Arg237Cys), p.(Gly448Asp), p.(Leu516Pro), and p.(Tyr582His) were characterized using bioinformatics, molecular modeling, and [35S]-sulfate uptake assays in Madin-Darby canine kidney cells.
    Results: All probands presented with concern for short stature and were found to have scoliosis and/or skeletal dysplasia. A reduction in plasma sulfate level and/or increase in urinary sulfate excretion was detected in 2 of 2 probands evaluated. Functional studies were consistent with SLC13A1 variants resulting in the complete loss of sulfate transport activity.
    Conclusion: Biallelic loss-of-function variants in SLC13A1 are a novel cause of skeletal phenotypes in humans with a measurable biomarker. Sulfate measurements should be considered in the clinical interpretation of variants identified in SLC13A1.
    Keywords:  Hyposulfatemia; Scoliosis; Short stature; Skeletal dysplasia; Sulfate transporter
    DOI:  https://doi.org/10.1016/j.gimo.2024.101958
  8. Animals (Basel). 2025 Jan 23. pii: 324. [Epub ahead of print]15(3):
    From Pre-Foaling to Late Pregnancy: Cortisol, DHEA(S), Progesterone, 17-β-Estradiol, and Allopregnanolone Hair Concentration Profiles in Standardbred Mares.
    Maria Cristina Veronesi, Alessio Cotticelli, Isabella Pividori, Matilde Giombolini, Mirco Corazzin, Letizia Ellero, Tanja Peric.
      Pregnancy and parturition represent two important physiologic phases in female mammals, in which metabolic, behavioral, and endocrinologic changes should occur in perfect timing and interaction, leading to a normal course of gestation and the occurrence of parturition at term, allowing the birth of mature and viable offspring. The present study aimed to describe the hormonal changes recorded in the hair occurring in mares from pre-foaling to late pregnancy. The hair cortisol (C) concentrations did not show any significant variations throughout the study, while dehydroepiandrosterone (sulfate) (DHEA(S)) hair concentrations showed an increase from ST-1 to ST4 (p < 0.01) followed by a significant decrease at ST6 (p < 0.01). From ST7 to ST8, hair DHEA(S) concentrations increased significantly (p < 0.01). In ST7 the cortisol/DHEA(S) ratio*100 was higher than in the other sampling times (p < 0.01) except for ST0, in which the ratio was similar to ST7. Hair progesterone (P4), 17-β-estradiol (E2), and allopregnanolone (AlloP) concentrations showed similar patterns through time with the exception of some differences between them at the end of the sampling period. The results showed that hormones fluctuations in the hair mimicked those observed previously in plasma making hair interesting for retrospective studies both on mares and foals during gestation without invasiveness and adding a complementary tool in studying the feto-maternal relationship.
    Keywords:  17-β-estradiol; DHEA(S); allopregnanolone; cortisol; hair; horse; late pregnancy; progesterone
    DOI:  https://doi.org/10.3390/ani15030324
  9. Int J Mol Sci. 2025 Feb 01. pii: 1273. [Epub ahead of print]26(3):
    mRNA Degradation as a Therapeutic Solution for Mucopolysaccharidosis Type IIIC: Use of Antisense Oligonucleotides to Promote Downregulation of Heparan Sulfate Synthesis.
    Juliana Inês Santos, Mariana Gonçalves, Matilde Barbosa Almeida, Hugo Rocha, Ana Joana Duarte, Liliana Matos, Luciana Vaz Moreira, Marisa Encarnação, Paulo Gaspar, Maria João Prata, Maria Francisca Coutinho, Sandra Alves.
      Mucopolysaccharidosis type IIIC is a neurodegenerative lysosomal storage disorder (LSD) characterized by the accumulation of undegraded heparan sulfate (HS) due to the lack of an enzyme responsible for its degradation: acetyl-CoA:α-glucosaminide N-acetyltransferase (HGSNAT). Classical treatments are ineffective. Here, we attempt a new approach in genetic medicine, genetic substrate reduction therapy (gSRT), to counteract this neurological disorder. Briefly, we used synthetic oligonucleotides, particularly gapmer antisense oligonucleotides (ASOs), to target the synthesis of the accumulated compounds at the molecular level, downregulating a specific gene involved in the first step of HS biosynthesis, XYLT1. Our goal was to reduce HS production and, consequently, its accumulation. Initially, five gapmer ASOs were designed and their potential to decrease XYLT1 mRNA levels were tested in patient-derived fibroblasts. Subsequent analyses focused on the two best performing molecules alone. The results showed a high inhibition of the XYLT1 gene mRNA (around 90%), a decrease in xylosyltransferase I (XT-I) protein levels and a reduction in HS storage 6 and 10 days after transfection (up to 21% and 32%, respectively). Overall, our results are highly promising and may represent the initial step towards the development of a potential therapeutic option not only for MPS IIIC, but virtually for every other MPS III form. Ultimately, the same principle may also apply to other neuropathic MPS.
    Keywords:  Lysosomal Storage Disorders (LSDs); Mucopolysaccharidosis type III (MPS III); RNA therapeutics; antisense oligonucleotides (ASOs); gapmer ASOs; genetic substrate reduction therapy (gSRT)
    DOI:  https://doi.org/10.3390/ijms26031273
  10. J Endocr Soc. 2025 Feb 04. 9(3): bvaf011
    Lower Cortisol and Dehydroepiandrosterone Sulfate and Higher Food Addiction in Childhood Obesity: Associations With Stress and Dietary Parameters.
    Rúbia Cartaxo Squizato de Moraes, Thallyta Alanna Ferreira Viana, Joicy Karla Grangeiro Pereira, Paulo César Trindade da Costa, Davyson Barbosa Duarte, Lydiane de Lima Tavares Toscano, Manuel Francisco de Araújo Lima, Melyssa Kellyane Cavalcanti Galdino, Joelma Rodrigues de Souza, Francisco Antônio de Oliveira Júnior, Adélia da Costa Pereira de Arruda Neta, José Luiz de Brito Alves, Vinícius José Baccin Martins.
       Context: Obesity has been associated with changes in cortisol and dehydroepiandrosterone (DHEA) sulfate concentrations and increased stress levels and food addiction.
    Objectives: We explored changes in morning salivary cortisol and DHEA in childhood obesity and their associations with body composition, metabolic profile, food addiction, food consumption, and stress in a cross-sectional study.
    Methods: Children aged 7 to 12 years of both sexes were allocated into 2 groups according to body mass index-for-age: control group (n = 60) or obesity group (n = 98). Anthropometric, body composition, serum glucose, insulin, lipid profile, and DHEA were measured. Saliva was collected at different times to measure morning salivary cortisol concentrations. Food addiction, food consumption, and stress were assessed using questionnaires.
    Results: Lower DHEA [1.04 (0.87-1.25) ng/mL vs 1.65 (1.30-2.07) ng/mL, P = .002] and salivary cortisol (6:00 Am: 1.17 ± 0.89 vs 1.45 ± 0.82 nmol/L, 6:30 Am: 1.53 ± 0.68 vs 1.83 ± 0.70 nmol/L, 7:30 Am: 0.72 ± 0.99 vs 1.31 ± 0.94 nmol/L, P-value of time < 0.001 and P-value of group = .002) were observed in children with obesity compared to the control. DHEA correlated negatively with waist circumference (r = -0.20, P < .05), body mass index-for-age(BMI-Z) (r = -0.21, P < .01), and weight (r = -0.25, P < .01). DHEA showed a positive correlation with the cortisol area under the curve (r = 0.29, P = .002). Food addiction was positively correlated with waist circumference (r = 0.21, P < .01), BMI-Z (r = 0.22, P < .01), body weight (r = 0.20, P < .05), total energy intake (r = 0.20, P < .05), and lipids (r = 0.24, P < .01).
    Conclusion: Children with obesity showed lower concentrations of salivary cortisol and DHEA and higher food addiction compared to control children. These changes may contribute to the development of chronic diseases over time.
    Keywords:  DHEA; NOVA classification; childhood obesity; cortisol; food addiction; stress
    DOI:  https://doi.org/10.1210/jendso/bvaf011
  11. J Anal Toxicol. 2025 Feb 13. pii: bkaf006. [Epub ahead of print]
    Blood to serum concentration ratios for ethyl glucuronide and ethyl sulfate after five drinking episodes.
    Matthias Bantle, Annette Thierauf-Emberger, Alexandra Schröck, Wolfgang Weinmann, Lorenz M Bell.
      Ethyl glucuronide (EtG) and ethyl sulfate (EtS) are mostly analyzed in urine; consequently, most kinetic studies are based on urine samples. In forensic cases, however, it may be necessary to determine these alcohol biomarkers in serum, whole blood or capillary blood. While there are sufficient data on EtG and EtS in serum after alcohol consumption, the amount of data available on whole blood concentrations is small. Therefore, data on corresponding blood-to-serum ratios seem to gain importance. This study provides data on a drinking experiment with 5 drinking episodes, where serum and whole blood samples were taken simultaneously from 11 healthy participants over 10 days. The samples were analyzed for EtG, EtS, and ethanol. EtG and EtS analysis in whole blood and serum were performed by LC -MS/MS; ethanol was determined by GC-FID and an ADH-based method. EtG and EtS reached their maximum concentration 4-7 hours after alcohol consumption. For EtG, a mean blood-to-serum ratio of 0.58 with a range from 0.38 to 0.73 was found; for EtS, the mean ratio was 0.81 with a range from 0.61 to 0.92, indicating a predominant distribution in the serum. For both analytes, high correlation coefficients were obtained when plotting concentrations in serum against concentrations in whole blood. Concerning elimination profiles of the individuals, no time or concentration dependence of EtG or EtS blood-to-serum ratios could be deduced. Neither for EtG nor for EtS a regularity of curve progressions could be observed in our test specimens.
    Keywords:  blood-to-serum ratio; ethanol; ethyl glucuronide; ethyl sulfate
    DOI:  https://doi.org/10.1093/jat/bkaf006
  12. Biosens Bioelectron. 2025 Jan 30. pii: S0956-5663(25)00066-1. [Epub ahead of print]275 117192
    Rational engineering of isoform-specific hSULT1E1 fluorogenic substrates for functional analysis and inhibitor screening.
    Xiaoting Niu, Yufan Fan, Guanghao Zhu, Hairong Zeng, Bei Zhao, Mengru Sun, Lin Chen, Luling Wu, Zhenhao Tian, Tony D James, Guangbo Ge.
      Human estrogen sulfotransferase (hSULT1E1), an important conjugative enzyme, plays crucial roles in both estrogen homeostasis and xenobiotic metabolism. Herein, a rational substrate engineering strategy was adopted to construct highly specific fluorogenic substrates for hSULT1E1. In the 1st round of structure-based virtual screening, 4-hydroxyl-1,8-naphthalimide (4-HN) was identified as a suitable scaffold for constructing hSULT1E1 substrates. Subsequently, structural modifications on the north part of 4-HN generated a panel of derivatives as substrate candidates, in which HN-299 was identified as a highly selective fluorogenic substrate for hSULT1E1. In the 3rd round of structural optimization, a "molecular growth" strategy on the south part of HN-299 was used to develop a highly selective and reactive substrate (HN-375). Under physiological conditions, HN-375 could be readily sulfated by hSULT1E1 to generate a single fluorescent product, which emitted bright green signals at around 510 nm and was fully identified as HN-375 4-O-sulfate (HNS). Further investigations indicated that HN-375 exhibited excellent isoform-specificity, rapid-response, ultrahigh sensitivity, and high signal-to-noise ratio, and as such was subsequently used for sensing SULT1E1 activity in hepatocellular carcinoma specimens and live organs. With HN-375 in hand, a practical fluorescence-based assay was established for high-throughput screening and characterization of hSULT1E1 inhibitors, as such two potent hSULT1E1 inhibitors were identified from in-house compound libraries. Collectively, this study showcases a groundbreaking strategy for engineering highly specific and sensitive fluorogenic substrates for target conjugative enzyme(s), while HN-375 emerges as a practical tool for sensing SULT1E1 activity in a biological context and for the high-throughput screening of inhibitors.
    Keywords:  Enzyme activity analysis; High-throughput screening; Human estrogen sulfotransferase (hSULT1E1); Structure-based substrate design
    DOI:  https://doi.org/10.1016/j.bios.2025.117192
  13. Plants (Basel). 2025 Jan 22. pii: 322. [Epub ahead of print]14(3):
    Non-Targeted Metabolome Analysis with Low-Dose Selenate-Treated Arabidopsis.
    Hongqiao Li, Tetsuya Mori, Rintaro Moriyama, Moeka Fujita, Genki Hatanaka, Naoki Shiotsuka, Ryota Hosomi, Akiko Maruyama-Nakashita.
      Selenate, the most common form of selenium (Se) in soil environments, is beneficial for higher plants. Selenate is similar to sulfate in terms of the structure and the manner of assimilation by plants, which involves the reduction of selenate to selenide and the replacement of an S moiety in the organic compounds such as amino acids. The nonspecific incorporation of seleno-amino acids into proteins induce Se toxicity in plants. Selenate alters the plant metabolism, particularly the S metabolism, which is comparable to the responses to S deficiency (-S). However, previous analyses involved high concentrations of selenate, and the effects of lower selenate doses have not been elucidated. In this study, we analyzed the metabolic changes induced by selenate treatment through a non-targeted metabolome analysis and found that 2 µM of selenate decreased the S assimilates and amino acids, and increased the flavonoids, while the glutathione levels were maintained. The results suggest that the decrease in amino acid levels, which is not detected under -S, along with the disruptions in S assimilation, amino acid biosynthesis pathways, and the energy metabolism, present the primary metabolic influences of selenate. These results suggest that selenate targets the energy metabolism and S assimilation first, and induces oxidative stress mitigation, represented by flavonoid accumulation, as a key adaptive response, providing a novel, possible mechanism in plant stress adaptation.
    Keywords:  amino acids; flavonoid; glucosinolates; non-targeted metabolome; selenate; selenium; sulfur metabolism
    DOI:  https://doi.org/10.3390/plants14030322
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