bims-supasi Biomed News
on Sulfation pathways and signalling
Issue of 2022–12–18
thirteen papers selected by
Jonathan Wolf Mueller, University of Birmingham
  1. Structural Characterization and Glycosaminoglycan Impurities Analysis of Chondroitin Sulfate from Chinese Sturgeon.
  2. Neuronal expression of ndst3 in early zebrafish development is responsive to Wnt signaling manipulation.
  3. Sulfation of Phenolic Acids: Chemoenzymatic vs. Chemical Synthesis.
  4. Nutrient niche specificity for glycosaminoglycans is reflected in polysaccharide utilization locus architecture of gut Bacteroides species.
  5. In Vitro Methods to Study the Golgi Apparatus Role in Proteoglycan and Glycosaminoglycan Synthesis.
  6. Introducing or removing heparan sulfate binding sites does not alter brain uptake of the blood-brain barrier shuttle scFv8D3.
  7. Distribution and postnatal development of chondroitin sulfate proteoglycans in the perineuronal nets of cholinergic motoneurons innervating extraocular muscles.
  8. Design and Validation of a Sensitive Multisteroid LC-MS/MS Assay for the Routine Clinical Use: One-Step Sample Preparation with Phospholipid Removal and Comparison to Immunoassays.
  9. Microbial synthesis of glycosaminoglycans and their oligosaccharides.
  10. Sleep and Anabolic/Catabolic Hormonal Profile in Sedentary Middle-Aged Adults: The FIT-AGEING Study.
  11. Unravelling microbial drivers of the sulfate-reduction process inside landfill using metagenomics.
  12. Serum uric acid/creatinine ratio and free androgen index are synergistically associated with increased risk of polycystic ovary syndrome in obese women.
  13. Impact of Sulfated Hyaluronan on Bone Metabolism in Diabetic Charcot Neuroarthropathy and Degenerative Arthritis.
  1. Polymers (Basel). 2022 Dec 05. pii: 5311. [Epub ahead of print]14(23):
    Structural Characterization and Glycosaminoglycan Impurities Analysis of Chondroitin Sulfate from Chinese Sturgeon.
    Mei Zhao, Yong Qin, Ying Fan, Xu Wang, Haixin Yi, Xiaoyu Cui, Fuchuan Li, Wenshuang Wang.
      Chinese sturgeon was an endangered cartilaginous fish. The success of artificial breeding has promoted it to a food fish and it is now beginning to provide a new source of cartilage for the extraction of chondroitin sulfate (CS). However, the structural characteristics of sturgeon CS from different tissues remain to be determined in more detail. In this study, CSs from the head, backbone, and fin cartilage of Chinese sturgeon were individually purified and characterized for the first time. The molecular weights, disaccharide compositions, and oligosaccharide sulfation patterns of these CSs are significantly different. Fin CS (SFCS), rich in GlcUAα1-3GalNAc(4S), has the biggest molecular weight (26.5 kDa). In contrast, head CS (SHCS) has a molecular weight of 21.0 kDa and is rich in GlcUAα1-3GalNAc(6S). Most features of backbone CS (SBCS) are between the former two. Other glycosaminoglycan impurities in these three sturgeon-derived CSs were lower than those in other common commercial CSs. All three CSs have no effect on the activity of thrombin or Factor Xa in the presence of antithrombin III. Hence, Chinese sturgeon cartilage is a potential source for the preparation of CSs with different features for food and pharmaceutical applications.
    Keywords:  Chinese sturgeon; chondroitin sulfate; disaccharide composition; molecular weight; oligosaccharides
    DOI:  https://doi.org/10.3390/polym14235311
  2. Gene Expr Patterns. 2022 Dec 08. pii: S1567-133X(22)00070-9. [Epub ahead of print] 119300
    Neuronal expression of ndst3 in early zebrafish development is responsive to Wnt signaling manipulation.
    Rebecca A Anderson, Usua Oyarbide.
      Heperan sulfate proteoglycans (HSPGs) are constituents of the cell surface and extracellular matrix and are vital for various activities within the cell. The N-deacetylase/N-sulfotransferase (heparin glucosaminyl) family of enzymes, or NDST, modifies heparan sulfate (HS) by catalyzing both the N-deacetylation and the N-sulfation of N-acetylglucosamine residues. In zebrafish, a single ndst3 gene is an orthologue of both mammalian NDST3 and NDST4 genes. The role of ndst3 in zebrafish development has not been investigated and such study may provide insight into the role(s) of both mammalian orthologous. Here, we characterized expression of ndst3 during early development in zebrafish and found it to be predominately neuronal. We found that expression of ndst3 is sensitive to Wnt signaling manipulation, with stimulation of the Wnt pathway resulting in robust expansion of ndst3 expression domains. Finally, using CRISPR/Cas9 genome editing, we mutagenized the ndst3 gene and isolated an allele, ndst3nu20, resulting in a frameshift and premature protein truncation. We discovered Ndst3 is not essential for zebrafish survival as ndst3nu20 homozygous mutants are viable and fertile.
    Keywords:  Heperan sulfate proteoglycans (HSPGs); NDST; Wnt signaling; Zebrafish; ndst3
    DOI:  https://doi.org/10.1016/j.gep.2022.119300
  3. Int J Mol Sci. 2022 Dec 02. pii: 15171. [Epub ahead of print]23(23):
    Sulfation of Phenolic Acids: Chemoenzymatic vs. Chemical Synthesis.
    Viola Kolaříková, Katerina Brodsky, Lucie Petrásková, Helena Pelantová, Josef Cvačka, Libor Havlíček, Vladimír Křen, Kateřina Valentová.
      Phenolic acids are known flavonoid metabolites, which typically undergo bioconjugation during phase II of biotransformation, forming sulfates, along with other conjugates. Sulfated derivatives of phenolic acids can be synthesized by two approaches: chemoenzymatically by 3'-phosphoadenosine-5'-phosphosulfate (PAPS)-dependent sulfotransferases or PAPS-independent aryl sulfotransferases such as those from Desulfitobacterium hafniense, or chemically using SO3 complexes. Both approaches were tested with six selected phenolic acids (2-hydroxyphenylacetic acid (2-HPA), 3-hydroxyphenylacetic acid (3-HPA), 4-hydroxyphenylacetic acid (4-HPA), 3,4-dihydroxyphenylacetic acid (DHPA), 3-(4-hydroxyphenyl)propionic acid (4-HPP), and 3,4-dihydroxyphenylpropionic acid (DHPP)) to create a library of sulfated metabolites of phenolic acids. The sulfates of 3-HPA, 4-HPA, 4-HPP, DHPA, and DHPP were all obtained by the methods of chemical synthesis. In contrast, the enzymatic sulfation of monohydroxyphenolic acids failed probably due to enzyme inhibition, whereas the same reaction was successful for dihydroxyphenolic acids (DHPA and DHPP). Special attention was also paid to the counterions of the sulfates, a topic often poorly reported in synthetic works. The products obtained will serve as authentic analytical standards in metabolic studies and to determine their biological activity.
    Keywords:  aryl sulfotransferase; biotransformation; flavonoid metabolites; phenolic acids; sulfation
    DOI:  https://doi.org/10.3390/ijms232315171
  4. Front Microbiol. 2022 ;13 1033355
    Nutrient niche specificity for glycosaminoglycans is reflected in polysaccharide utilization locus architecture of gut Bacteroides species.
    Annelieke Overbeeke, Bela Hausmann, Georgi Nikolov, Fatima C Pereira, Craig W Herbold, David Berry.
       Introduction: Glycosaminoglycans (GAGs) present in the mucosal layer can be used as nutrients by certain intestinal bacteria, particularly members of the Bacteroides. GAG abundances are altered in some diseases such as inflammatory bowel diseases, which may affect microbial composition and activity, and it is therefore important to understand GAG utilization by members of the gut microbiota.
    Methods: We used growth assays, transcriptomics, and comparative genomics to evaluate chondroitin sulfate (CS) and hyaluronan (HA) degradation ability by multiple gut Bacteroides species.
    Results and discussion: We found that not all Bacteroides species able to degrade CS could also degrade HA, despite having lyases which act on both compounds. We propose that in the model organism Bacteroides thetaiotaomicron, the lyase BT_3328 in combination with surface binding proteins BT_3329 and BT_3330 and potentially BT_4411 are involved in HA breakdown. Furthermore, degradation of both compounds provides public goods for other Bacteroides, including non-degraders, suggesting that cooperative degradation as well as cross-feeding may be widespread in the mucosal glycan utilization clade.
    Keywords:  bacteroides; chondroitin sulfate; glycosaminoglycans; hyaluronan; intestine
    DOI:  https://doi.org/10.3389/fmicb.2022.1033355
  5. Methods Mol Biol. 2023 ;2557 709-720
    In Vitro Methods to Study the Golgi Apparatus Role in Proteoglycan and Glycosaminoglycan Synthesis.
    Kristian Prydz.
      Subcellular fractionation is an introductory step in a variety of experimental approaches designed to study intracellular components, like membranes and organelle systems. Subcellular fractions enriched in membranes of the Golgi apparatus of mammalian cells have been isolated to address localization and activity of proteins, including enzymes, to study intracellular membrane transport mechanisms, and to reconstitute in vitro cellular processes associated with the Golgi apparatus. Here, I describe methods to purify Golgi membranes by subcellular fractionation, to assay nucleotide sulfate (PAPS) uptake into Golgi vesicles, and to measure sulfate incorporation into in vitro synthesized glycosaminoglycans.
    Keywords:  3′-Phosphoadenosine-5′-phosphosulfate (PAPS); Glycosaminoglycans (GAGs); Golgi apparatus; Proteoglycans (PGs); Sucrose gradients; Sulfation; Ultracentrifugation
    DOI:  https://doi.org/10.1007/978-1-0716-2639-9_42
  6. Sci Rep. 2022 Dec 12. 12(1): 21479
    Introducing or removing heparan sulfate binding sites does not alter brain uptake of the blood-brain barrier shuttle scFv8D3.
    Andrés de la Rosa, Nicole G Metzendorf, Jamie I Morrison, Rebecca Faresjö, Fadi Rofo, Alex Petrovic, Paul O'Callaghan, Stina Syvänen, Greta Hultqvist.
      The blood-brain barrier (BBB) greatly limits the delivery of protein-based drugs into the brain and is a major obstacle for the treatment of brain disorders. Targeting the transferrin receptor (TfR) is a strategy for transporting protein-based drugs into the brain, which can be utilized by using TfR-binding BBB transporters, such as the TfR-binding antibody 8D3. In this current study, we investigated if binding to heparan sulfate (HS) contributes to the brain uptake of a single chain fragment variable of 8D3 (scFv8D3). We designed and produced a scFv8D3 mutant, engineered with additional HS binding sites, HS(+)scFv8D3, to assess whether increased HS binding would improve brain uptake. Additionally, a mutant with a reduced number of HS binding sites, HS(-)scFv8D3, was also engineered to see if reducing the HS binding sites could also affect brain uptake. Heparin column chromatography showed that only the HS(+)scFv8D3 mutant bound HS in the experimental conditions. Ex vivo results showed that the brain uptake was unaffected by the introduction or removal of HS binding sites, which indicates that scFv8D3 is not dependent on the HS binding sites for brain uptake. Conversely, introducing HS binding sites to scFv8D3 decreased its renal excretion while removing them had the opposite effect.
    DOI:  https://doi.org/10.1038/s41598-022-25965-x
  7. Sci Rep. 2022 Dec 14. 12(1): 21606
    Distribution and postnatal development of chondroitin sulfate proteoglycans in the perineuronal nets of cholinergic motoneurons innervating extraocular muscles.
    Adrienn Ritok, Peter Kiss, Anas Zaher, Ervin Wolf, Laszlo Ducza, Timea Bacskai, Clara Matesz, Botond Gaal.
      Fine control of extraocular muscle fibers derives from two subpopulations of cholinergic motoneurons in the oculomotor-, trochlear- and abducens nuclei. Singly- (SIF) and multiply innervated muscle fibers (MIF) are supplied by the SIF- and MIF motoneurons, respectively, representing different physiological properties and afferentation. SIF motoneurons, as seen in earlier studies, are coated with chondroitin sulfate proteoglycan rich perineuronal nets (PNN), whereas MIF motoneurons lack those. Fine distribution of individual lecticans in the composition of PNNs and adjacent neuropil, as well as the pace of their postnatal accumulation is, however, still unknown. Therefore, the present study aims, by using double immunofluorescent identification and subsequent morphometry, to describe local deposition of lecticans in the perineuronal nets and neuropil of the three eye movement nuclei. In each nucleus PNNs were consequently positive only with WFA and aggrecan reactions, suggesting the dominating role of aggrecan is PNN establishment. Brevican, neurocan and versican however, did not accumulate at all in PNNs but were evenly and moderately present throughout the neuropils. The proportion of PNN bearing motoneurons appeared 76% in oculomotor-, 72.2% in trochlear- and 78.3% in the abducens nucleus. We also identified two morphological subsets of PNNs, the focal and diffuse nets of SIF motoneurons. The process of CSPG accumulation begins just after birth, although considerable PNNs occur at week 1 age around less than half of the motoneurons, which ratio doubles until 2-month age. These findings may be related to the postnatal establishment of the oculokinetic network, performing different repertoires of voluntary eye movements in functionally afoveolate and foveolate animals.
    DOI:  https://doi.org/10.1038/s41598-022-25692-3
  8. Int J Mol Sci. 2022 Nov 24. pii: 14691. [Epub ahead of print]23(23):
    Design and Validation of a Sensitive Multisteroid LC-MS/MS Assay for the Routine Clinical Use: One-Step Sample Preparation with Phospholipid Removal and Comparison to Immunoassays.
    Valentin Braun, Hermann Stuppner, Lorenz Risch, Christoph Seger.
      Steroid analysis in clinical laboratories is dominated by immunoassays (IAs) that have a high sample turnover but are inherently limited in trueness, precision, and sensitivity. Liquid chromatography coupled to mass spectrometry (LC-MS/MS) has proved to be a far more capable tool, delivering better sensitivity, specificity, and the possibility of parallel analysis of multiple steroids and metabolites, providing the endocrinologist with more reliable and comprehensive diagnostic information. An LC-MS/MS assay with gradient elution over less than eight minutes and a one-step sample preparation combining protein precipitation with phospholipid removal of off-line solid-phase extraction was developed and validated. It allowed the quantification of 11-deoxycorticosterone (11-DOC), 11-deoxycortisol (11-DF), 17-OH-progesterone (17P), 21-deoxycortisol (21-DF), androstenedione (ANDRO), aldosterone (ALDO), corticosterone (CC), cortisol (CL), cortisone (CN), dehydroepiandrosterone (DHEA), dehydroepiandrosterone sulfate (DHEAS), dihydrotestosterone (DHT), estradiol (E2), progesterone (PROG), and testosterone (TES) in human serum. Interday imprecision was generally better than 15%, trueness was proven by recovery experiments with ISO 17034-certified reference materials, proficiency testing (UK NEQAS), and measuring serum reference standards. In-house comparison against IVD-CE-certified immunoassays (IA) for 17P, ANDRO, CL, DHEAS, E2, PROG, and TES was conducted by assessing leftover routine patient samples and purpose-built patient serum pools. None of the compared routine IAs were meeting the standards of the LC-MS/MS. Insufficient overall comparability was found for ANDRO and 17P (mean bias > +65%). Accuracy limitations at lower concentrations were present in IAs for PROG, E2, and TES.
    Keywords:  LC-MS/MS; endocrinology; laboratory medicine; steroid measurement
    DOI:  https://doi.org/10.3390/ijms232314691
  9. Trends Microbiol. 2022 Dec 13. pii: S0966-842X(22)00311-0. [Epub ahead of print]
    Microbial synthesis of glycosaminoglycans and their oligosaccharides.
    Kaifang Liu, Liang Guo, Xiulai Chen, Liming Liu, Cong Gao.
      Compared with chemical synthesis and tissue extraction methods, microbial synthesis of glycosaminoglycans (GAGs) is attractive because of the advantages of eco-friendly processes, production safety, and sustainable development. However, boosting the efficiency of microbial cell factories, precisely regulating GAG molecular weights, and rationally controlling the sulfation degree of GAGs remain challenging. To address these issues, various strategies, including genetic, enzymatic, metabolic, and fermentation engineering, have been developed. In this review, we summarize the recent progress in the construction of efficient GAG-producing microbial cell factories, regulation of the molecular weight of GAGs, and modification of GAG chains. Moreover, future studies, remaining challenges, and potential solutions in this field are discussed.
    Keywords:  glycosaminoglycans; microbial engineering; molecular weight regulation; sulfated modification; synthetic metabolism
    DOI:  https://doi.org/10.1016/j.tim.2022.11.003
  10. Int J Mol Sci. 2022 Nov 25. pii: 14709. [Epub ahead of print]23(23):
    Sleep and Anabolic/Catabolic Hormonal Profile in Sedentary Middle-Aged Adults: The FIT-AGEING Study.
    Sol Mochón-Benguigui, Almudena Carneiro-Barrera, Manuel Dote-Montero, Manuel J Castillo, Francisco J Amaro-Gahete.
      Sleep quality plays an important role in the modulation of several aging markers. This influence could be explained by aging-induced hormonal changes. Indeed, poor sleep quality has been associated with the development of several endocrine-related health complications. This study examined the relationship of both subjective and objective sleep quantity and quality, with basal levels of selected plasma anabolic and catabolic hormones in sedentary middle-aged adults. A total of 74 volunteers (52.7% women; aged 53.7 ± 5.1) were recruited for this study. Subjective sleep quality was assessed by the Pittsburgh Sleep Quality Index (PSQI; higher scores indicate worse sleep quality), and objective sleep quality parameters (total sleep time [TST], wake after sleep onset [WASO], and sleep efficiency [SE]) were measured using a wrist-worn accelerometer. Basal levels of plasma dehydroepiandrosterone sulphate (DHEAS), total testosterone, sex hormone binding globulin (SHBG), somatotropin, and cortisol levels, were determined. Free testosterone was calculated from the total testosterone and SHBG levels. No associations of global PSQI score, TST, WASO, and SE with DHEAS, free testosterone, and somatotropin plasma levels were found, neither in men nor in women (all p ≥ 0.05). Global PSQI score was inversely related to cortisol plasma levels in women (p = 0.043). WASO was positively associated with cortisol plasma levels, while SE was negatively associated with cortisol plasma levels in women (all p ≤ 0.027). Sleep quality is not related to levels of plasma anabolic hormones, but to levels of catabolic hormones, in sedentary middle-aged adults. Therefore, these results suggest that potential changes in aging biomarkers associated with sleep disturbances, could be mediated by age-related changes in the catabolic endocrine system.
    Keywords:  actigraphy; cortisol; dehydroepiandrosterone sulphate; hormone; somatotropin; testosterone
    DOI:  https://doi.org/10.3390/ijms232314709
  11. Chemosphere. 2022 Dec 12. pii: S0045-6535(22)04030-9. [Epub ahead of print] 137537
    Unravelling microbial drivers of the sulfate-reduction process inside landfill using metagenomics.
    Lifang Hu, Yuqian Wang, Manting Ci, Yuyang Long.
      Hydrogen sulfide (H2S) is one of the common landfill odor. This research demonstrates that the sulfate transformation behavior is significantly enhanced during the landfill process, accompanied by a shift in microbial structure. The relative abundance of dissimilatory sulfate reduction (DSR) and thiosulfate oxidation by SOX (sulfur-oxidation) complex gradually decreases through the landfill processes while the assimilatory sulfate reduction (ASR) demonstrates the opposite behavior. The major module for landfill sulfate reduction is ASR, accounting for 31.72% ± 2.84% of sulfate metabolism. Based on the functional genes for the sulfate pathway, the drivers for sulfate biotransformation in landfills were determined and further identified their contribution in the sulfate metabolism during landfill processes. Pseudomonas, Methylocaldum, Bacillus, Methylocystis and Hyphomicrobium were the top 5 contributors for ASR pathway, and only one genus Pseudomonas was found for DSR pathway. Among the 26 high-quality metagenome-assembled genomes of sulfate functional species, 24 were considered novel species for sulfuric metabolism. Overall, this study provides unique insight into the sulfate transformation process related to the H2S odor control in landfill management.
    Keywords:  ASR; Landfill; Metagenome-assembled genomes; SRB; Sulfur cycling
    DOI:  https://doi.org/10.1016/j.chemosphere.2022.137537
  12. BMC Endocr Disord. 2022 Dec 13. 22(1): 315
    Serum uric acid/creatinine ratio and free androgen index are synergistically associated with increased risk of polycystic ovary syndrome in obese women.
    Mervat M El-Eshmawy, Asmaa Ibrahim, Rania Bahriz, Nermeen Shams-Eldin, Nancy Mahsoub.
       BACKGROUND: Features of metabolic syndrome such as abdominal obesity, insulin resistance, hypertension and dyslipidemia are commonly encountered in polycystic ovary syndrome (PCOS). Recent evidence has suggested an association between high serum uric acid/creatinine (UA/Cr) ratio and metabolic syndrome however, no studies have investigated this association in PCOS. The current study was conducted to investigate the relationship between UA/Cr ratio and PCOS and to identify whether UA/Cr ratio and free androgen index (FAI) have an additive interaction for detection of PCOS risk in obese women.
    METHODS: This study enrolled 40 obese women with PCOS and 40 control women with regular menstrual cycles matched for age and body mass index (BMI). Anthropometric measurements, fasting glucose, fasting insulin, homeostasis model assessment of insulin resistance (HOMA-IR), lipids profile, luteinizing hormone (LH), follicle stimulating hormone (FSH), estradiol, dehydroepiandrosterone sulfate (DHEAS), sex hormone binding globulin (SHBG), total testosterone, free androgen index (FAI), UA/Cr ratio were assessed.
    RESULTS: Serum UA/Cr ratio was significantly higher in obese women with PCOS than in non-PCOS women. UA/Cr ratio was correlated with BMI, waist and neck circumferences, blood pressure, fasting insulin, HOMA-IR, lipids, LH/FSH, estradiol, DHEAS, total testosterone, FAI and SHBG. UA/Cr ratio and FAI were independent risk factors for PCOS in obese women however, the addictive interaction between UA/Cr ratio and FAI had a higher fold risk (OR: 4.3, 95% CI, 3.4-7.58) and a more significance (P = 0.002) for determination of PCOS.
    CONCLUSION: Serum UA/Cr ratio combined with FAI can exert an additive or synergistic impact on prediction of PCOS in obese women.
    Keywords:  Free androgen index; Polycystic ovary syndrome; Serum uric acid/creatinine ratio
    DOI:  https://doi.org/10.1186/s12902-022-01240-y
  13. Int J Mol Sci. 2022 Dec 02. pii: 15146. [Epub ahead of print]23(23):
    Impact of Sulfated Hyaluronan on Bone Metabolism in Diabetic Charcot Neuroarthropathy and Degenerative Arthritis.
    Sabine Schulze, Christin Neuber, Stephanie Möller, Ute Hempel, Lorenz C Hofbauer, Klaus-Dieter Schaser, Jens Pietzsch, Stefan Rammelt.
      Bone in diabetes mellitus is characterized by an altered microarchitecture caused by abnormal metabolism of bone cells. Together with diabetic neuropathy, this is associated with serious complications including impaired bone healing culminating in complicated fractures and dislocations, especially in the lower extremities, so-called Charcot neuroarthropathy (CN). The underlying mechanisms are not yet fully understood, and treatment of CN is challenging. Several in vitro and in vivo investigations have suggested positive effects on bone regeneration by modifying biomaterials with sulfated glycosaminoglycans (sGAG). Recent findings described a beneficial effect of sGAG for bone healing in diabetic animal models compared to healthy animals. We therefore aimed at studying the effects of low- and high-sulfated hyaluronan derivatives on osteoclast markers as well as gene expression patterns of osteoclasts and osteoblasts from patients with diabetic CN compared to non-diabetic patients with arthritis at the foot and ankle. Exposure to sulfated hyaluronan (sHA) derivatives reduced the exaggerated calcium phosphate resorption as well as the expression of genes associated with bone resorption in both groups, but more pronounced in patients with CN. Moreover, sHA derivatives reduced the release of pro-inflammatory cytokines in osteoclasts of patients with CN. The effects of sHA on osteoblasts differed only marginally between patients with CN and non-diabetic patients with arthritis. These results suggest balancing effects of sHA on osteoclastic bone resorption parameters in diabetes.
    Keywords:  Charcot neuroarthropathy; ankle arthritis; diabetes mellitus; osteoblasts; osteoclasts; sulfated hyaluronan
    DOI:  https://doi.org/10.3390/ijms232315146
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