bims-supasi 2023-09-24 papers

bims-supasi

Biomed News

on Sulfation pathways and signalling

Issue of 2023–09–24
eight papers selected by
Jonathan Wolf Mueller, University of Birmingham

Branch distribution pattern and anticoagulant activity of a fucosylated chondroitin sulfate from Phyllophorella kohkutiensis.
Structure-function of DHEA binding proteins.
Host heparan sulfate promotes ACE2 super-cluster assembly and enhances SARS-CoV-2-associated syncytium formation.
Sex steroid hormones and allergic diseases in children: a pilot birth cohort study in the Japan Environment and Children's Study cohort.
Analysis of progesterone and estrone-sulfate in feces of American Bison using liquid chromatography coupled to mass spectrometry: Technical validation and correlation with blood levels.
Immobilised-enzyme microreactors for the identification and synthesis of conjugated drug metabolites.
Structure of human drug transporters OATP1B1 and OATP1B3.
A novel monoclonal antibody against 6-sulfo sialyl Lewis x glycans attenuates murine allergic rhinitis by suppressing Th2 immune responses.

Carbohydr Polym. 2023 Dec 01. pii: S0144-8617(23)00769-5. [Epub ahead of print]321 121304

Branch distribution pattern and anticoagulant activity of a fucosylated chondroitin sulfate from Phyllophorella kohkutiensis.

Di Lan, Jiali Zhang, Xiaolei Shang, Lijuan Yu, Chen Xu, Pin Wang, Lige Cui, Nanqi Cheng, Huifang Sun, Jianing Ran, Le Sha, Ronghua Yin, Na Gao, Jinhua Zhao.

  Fucosylated chondroitin sulfate (FCS) extracted from Phyllophorella kohkutiensis (PkFCS) is composed of d-GalNAc, d-GlcA, l-Fuc and -SO42-. According to the defined structures revealed by NMR spectra of the branches released by mild acid hydrolysis and oligosaccharides generated by β-eliminative depolymerization, the backbone of PkFCS is CS-E, and the branch types attached to C-3 of d-GlcA include l-Fuc2S4S, l-Fuc3S4S, l-Fuc4S, and the disaccharide α-d-GalNAc-1,2-α-l-Fuc3S4S with the ratio of 43:13:22:22. Notably, novel heptasaccharide and hendecasaccharide were identified that are branched with continuous distribution of the disaccharide. The structural sequences of the oligosaccharides indicate that three unique structural motifs are present in the entire PkFCS polymer, including a motif branched with randomly distributed different sulfated l-Fuc units, a motif containing regular l-Fuc2S4S branches and a motif enriched in α-d-GalNAc-1,2-α-l-Fuc3S4S. This is the first report about the distribution pattern of diverse branches in natural FCS. Natural PkFCS exhibited potent anticoagulant activity on APTT prolonging and anti-iXase activity. Regarding the structurally defined oligosaccharides with sulfated fucosyl side chains, octasaccharide (Pk4b) is the minimum fragment responsible for its anticoagulant activity correlated with anti-iXase. However, further glycosyl modification with a non-sulfated d-GalNAc at the C-2 position of l-Fuc3S4S could significantly decrease the anticoagulant and anti-iXase activity.

Keywords:  Anticoagulant; Disaccharide branch; Distribution pattern; Fucosylated chondroitin sulfate; Oligosaccharides; iXase

DOI:  https://doi.org/10.1016/j.carbpol.2023.121304
Vitam Horm. 2023 ;pii: S0083-6729(22)00092-9. [Epub ahead of print]123 587-617

Structure-function of DHEA binding proteins.

Barbara J Clark, Carolyn M Klinge.

  Dehydroepiandrosterone (3β-hydroxy-5-androsten-17-one, DHEA) and its sulfated metabolite DHEA-S are the most abundant circulating steroids and are precursors for active sex steroid hormones, estradiol and testosterone. DHEA has a broad range of reported effects in the central nervous system (CNS), cardiovascular system, adipose tissue, kidney, liver, and in the reproductive system. The mechanisms by which DHEA and DHEA-S initiate their biological effects are diverse. DHEA and DHEA-S may directly bind to plasma membrane (PM) receptors, including a DHEA-specific, G-protein coupled receptor (GPCR) in endothelial cells; various neuroreceptors, e.g., aminobutyric-acid-type A (GABA(A)), N-methyl-d-aspartate (NMDA) and sigma-1 (S1R) receptors (NMDAR and SIG-1R). DHEA and DHEA-S directly bind the nuclear androgen and estrogen receptors (AR, ERα, or ERβ) although with significantly lower binding affinities compared to the steroid hormones, e.g., testosterone, dihydrotestosterone, and estradiol, which are the cognate ligands for AR and ERs. Thus, extra-gonadal metabolism of DHEA to the sex hormones must be considered for many of the biological benefits of DHEA. DHEA also actives GPER1 (G protein coupled estrogen receptor 1). DHEA activates constitutive androstane receptor CAR (CAR) and proliferator activated receptor (PPARα) by indirect dephosphorylation. DHEA affects voltage-gated sodium and calcium ion channels and DHEA-2 activates TRPM3 (Transient Receptor Potential Cation Channel Subfamily M Member 3). This chapter updates our previous 2018 review pertaining to the physiological, biochemical, and molecular mechanisms of DHEA and DHEA-S activity.

Keywords:  DHEA; DHEA-S; Ion channels; Neurosteroid; Nuclear receptors; Receptors; Review; Signal transduction

DOI:  https://doi.org/10.1016/bs.vh.2022.12.002
Nat Commun. 2023 09 18. 14(1): 5777

Host heparan sulfate promotes ACE2 super-cluster assembly and enhances SARS-CoV-2-associated syncytium formation.

Qi Zhang, Weichun Tang, Eduardo Stancanelli, Eunkyung Jung, Zulfeqhar Syed, Vijayakanth Pagadala, Layla Saidi, Catherine Z Chen, Peng Gao, Miao Xu, Ivan Pavlinov, Bing Li, Wenwei Huang, Liqiang Chen, Jian Liu, Hang Xie, Wei Zheng, Yihong Ye.

SARS-CoV-2 infection causes spike-dependent fusion of infected cells with ACE2 positive neighboring cells, generating multi-nuclear syncytia that are often associated with severe COVID. To better elucidate the mechanism of spike-induced syncytium formation, we combine chemical genetics with 4D confocal imaging to establish the cell surface heparan sulfate (HS) as a critical stimulator for spike-induced cell-cell fusion. We show that HS binds spike and promotes spike-induced ACE2 clustering, forming synapse-like cell-cell contacts that facilitate fusion pore formation between ACE2-expresing and spike-transfected human cells. Chemical or genetic inhibition of HS mitigates ACE2 clustering, and thus, syncytium formation, whereas in a cell-free system comprising purified HS and lipid-anchored ACE2, HS stimulates ACE2 clustering directly in the presence of spike. Furthermore, HS-stimulated syncytium formation and receptor clustering require a conserved ACE2 linker distal from the spike-binding site. Importantly, the cell fusion-boosting function of HS can be targeted by an investigational HS-binding drug, which reduces syncytium formation in vitro and viral infection in mice. Thus, HS, as a host factor exploited by SARS-CoV-2 to facilitate receptor clustering and a stimulator of infection-associated syncytium formation, may be a promising therapeutic target for severe COVID.

DOI: https://doi.org/10.1038/s41467-023-41453-w
BMC Pediatr. 2023 Sep 21. 23(1): 479

Sex steroid hormones and allergic diseases in children: a pilot birth cohort study in the Japan Environment and Children's Study cohort.

Japan Environment and Children’s Study (JECS)

   BACKGROUND: Numerous studies suggest that sex steroids might play a role in sex disparity observed in allergic diseases in adults. However, whether sex hormones influence allergic diseases in children remains unclear. The aim of the present study was to examine the association of sex steroid hormones with allergic disease in Japanese children.
METHODS: The present cross-sectional study included 145 6-year-old children participating in a pilot birth cohort study in the Japan Environment and Children's Study. Data on allergic diseases were obtained from questionnaires, and serum levels of sex steroid hormones and allergen-specific IgE were measured. Logistic regression was performed to evaluate the association of sex hormones with allergic diseases.
RESULTS: After adjusted sex, amount of body fat at 6 years, parental history of allergic disease, and exposure to tobacco smoke, serum dehydroepiandrosterone sulfate level was significantly associated with reduced odds of any allergic disease (adjusted odds ratio, 0.58; 95% confidence interval, 0.36-0.93; P = 0.024) and serum follicle-stimulating hormone level was significantly associated with increased odds of any allergic disease (adjusted odds ratio, 2.04; 95% confidence interval, 1.01-4.11, P = 0.046). Dehydroepiandrosterone sulfate level showed a significant association with number of allergic diseases.
CONCLUSIONS: The current study findings suggest that sex hormones may play an important role in the development of allergic diseases in prepubertal children.

Keywords:  Allergic disease; Asthma; Atopic dermatitis; Dehydroepiandrosterone sulfate; Follicle-stimulating hormone; Sex steroid hormone

DOI:  https://doi.org/10.1186/s12887-023-04302-9
Domest Anim Endocrinol. 2023 Aug 30. pii: S0739-7240(23)00035-8. [Epub ahead of print]86 106819

Analysis of progesterone and estrone-sulfate in feces of American Bison using liquid chromatography coupled to mass spectrometry: Technical validation and correlation with blood levels.

Patrice Dufour, Vincent Frisée, Goulven Rigaux, Flore Brutinel, Sophie Egyptien, Philippe Bossaert, Jessica Deleersnyder, Stéfan Deleuze, Stéphanie Peeters, Caroline Le Goff, Jérôme Ponthier, Etienne Cavalier.

  American Bison's wild nature limits blood sample availability to study its endocrinology. This report describes progesterone (P4) and estrone-sulfate (E1S) assays in American Bison feces using Liquid Chromatography coupled with Mass Spectrometry (LC-MS). In 2 ranches, samples of feces (n = 73) and serum (n = 93) were collected in pregnant and nonpregnant American Bison. Feces samples (250 mg) were extracted with methanol, purified, and concentrated. Then, feces and serum samples were assayed using LC-MS, according to our previously described technique. Fecal matrix homogeneity was determined by measuring steroids in different areas of the sample and concentration evolutions were evaluated after storage at room temperature. During the validation process, lower limits of quantification were 20 pg/g (E1S) and 4 ng/g (P4) by meeting the following criteria: relative standard deviation <15% and relative bias <15%. By measuring hormones in different spots from the same sample, a moderate variability for E1S (coefficient of variation [CV] up to 21.3%) and a high variability for P4 (CV up to 85.5%) were highlighted. Correlation between concentrations in feces and in serum was higher for E1S (r = 0.77) than for P4 (r = 0.65) and P4 could be assayed in pregnant and nonpregnant animals whereas E1S was only present in pregnant. Feces storage at room temperature induced modification of steroid concentrations. The quantification of E1S and, at a lower level, of P4 in feces is an interesting alternative to serum assay to describe the pregnancy-related evolution of these steroids in American Bisons, with feces ideally stored frozen and mixed before the LC-MS procedures.

Keywords:  Bison Bison; Estrone-sulfate; Feces; Liquid-chromatography and mass spectrometry; Pregnancy; Progesterone

DOI:  https://doi.org/10.1016/j.domaniend.2023.106819
RSC Adv. 2023 Sep 18. 13(40): 27696-27704

Immobilised-enzyme microreactors for the identification and synthesis of conjugated drug metabolites.

Bradley Doyle, Leigh A Madden, Nicole Pamme, Huw S Jones.

The study of naturally circulating drug metabolites has been a focus of interest, since these metabolites may have different therapeutic and toxicological effects compared to the parent drug. The synthesis of metabolites outside of the human body is vital in order to conduct studies into the pharmacological activities of drugs and bioactive compounds. Current synthesis methods require significant purification and separation efforts or do not provide sufficient quantities for use in pharmacology experiments. Thus, there is a need for simple methods yielding high conversions whilst bypassing the requirement for a separation. Here we have developed and optimised flow chemistry methods in glass microfluidic reactors utilising surface-immobilised enzymes for sulfonation (SULT1a1) and glucuronidation (UGT1a1). Conversion occurs in flow, the precursor and co-factor are pumped through the device, react with the immobilised enzymes and the product is then simply collected at the outlet with no separation from a complex biological matrix required. Conversion only occurred when both the correct co-factor and enzyme were present within the microfluidic system. Yields of 0.97 ± 0.26 μg were obtained from the conversion of resorufin into resorufin sulfate over 2 h with the SULT1a1 enzyme and 0.47 μg of resorufin glucuronide over 4 h for UGT1a1. This was demonstrated to be significantly more than static test tube reactions at 0.22 μg (SULT1a1) and 0.19 μg (UGT1a1) over 4 h. With scaling out and parallelising, useable quantities of hundreds of micrograms for use in pharmacology studies can be synthesised simply.

DOI: https://doi.org/10.1039/d3ra03742h
Nat Commun. 2023 09 18. 14(1): 5774

Structure of human drug transporters OATP1B1 and OATP1B3.

Anca-Denise Ciută, Kamil Nosol, Julia Kowal, Somnath Mukherjee, Ana S Ramírez, Bruno Stieger, Anthony A Kossiakoff, Kaspar P Locher.

The organic anion transporting polypeptides OATP1B1 and OATP1B3 are membrane proteins that mediate uptake of drugs into the liver for subsequent conjugation and biliary excretion, a key step in drug elimination from the human body. Polymorphic variants of these transporters can cause reduced drug clearance and adverse drug effects such as statin-induced rhabdomyolysis, and co-administration of OATP substrates can lead to damaging drug-drug interaction. Despite their clinical relevance in drug disposition and pharmacokinetics, the structure and mechanism of OATPs are unknown. Here we present cryo-EM structures of human OATP1B1 and OATP1B3 bound to synthetic Fab fragments and in functionally distinct states. A single estrone-3-sulfate molecule is bound in a pocket located in the C-terminal half of OATP1B1. The shape and chemical nature of the pocket rationalize the preference for diverse organic anions and allow in silico docking of statins. The structure of OATP1B3 is determined in a drug-free state but reveals a bicarbonate molecule bound to the conserved signature motif and a histidine residue that is prevalent in OATPs exhibiting pH-dependent activity.

DOI: https://doi.org/10.1038/s41467-023-41552-8
Sci Rep. 2023 Sep 21. 13(1): 15740

A novel monoclonal antibody against 6-sulfo sialyl Lewis x glycans attenuates murine allergic rhinitis by suppressing Th2 immune responses.

Wei Liu, Wei Xiong, Wenxin Liu, Jotaro Hirakawa, Hiroto Kawashima.

Lymphocyte homing is mediated by the interaction between L-selectin on lymphocytes and its glycoprotein ligands modified with 6-sulfo sialyl Lewis x (6-sulfo sLex) glycans on high endothelial venules (HEVs) in peripheral lymph nodes (PLNs). However, the lack of specific antibodies reactive with both human and mouse 6-sulfo sLex has limited our understanding of its function in vivo. Here, we generated a novel monoclonal antibody, termed SF1, that specifically reacts with 6-sulfo sLex expressed on HEVs in both species in a manner dependent on sulfate, fucose, and sialic acid modifications. Glycan array and biolayer interferometry analyses indicated that SF1 specifically bound to 6-sulfo sLex with a dissociation constant of 6.09 × 10-9 M. SF1 specifically bound to four glycoproteins from PLNs corresponding to the molecular sizes of L-selectin ligand glycoproteins. Consistently, SF1 inhibited L-selectin-dependent lymphocyte rolling on 6-sulfo sLex-expressing cells ex vivo and lymphocyte homing to PLNs and nasal-associated lymphoid tissues in vivo. Furthermore, SF1 significantly attenuated ovalbumin-induced allergic rhinitis in mice in association with significant suppression of Th2 immune responses. Collectively, these results suggest that SF1 can be useful for the functional analysis of 6-sulfo sLex and may potentially serve as a novel therapeutic agent against immune-related diseases.

DOI: https://doi.org/10.1038/s41598-023-43017-w