bims-supasi 2023-04-09 papers

bims-supasi

Biomed News

on Sulfation pathways and signalling

Issue of 2023–04–09
nine papers selected by
Jonathan Wolf Mueller, University of Birmingham

Apolipoprotein E Recognizes Alzheimer's Disease Associated 3-O Sulfation of Heparan Sulfate.
Heparan sulfate glycomimetics via iterative assembly of "clickable" disaccharides.
Molecular weight determination of low molecular weight hyaluronic acid and chondroitin sulfate by gel permeation chromatography.
Combined treatment with glucosamine and chondroitin sulfate improves rheumatoid arthritis in rats by regulating the gut microbiota.
Absolute pharmacokinetics of heparin in primates.
Cholesterol sulfate limits neutrophil recruitment and gut inflammation during mucosal injury.
Effects of structural characteristics of (un)conjugated steroid metabolites in their collision cross section value.
Nitric oxide and hydrogen peroxide mediated regulation of chromium (VI) toxicity in wheat seedlings involves alterations in antioxidants and high affinity sulfate transporter.
Glycosaminoglycans: What Remains To Be Deciphered?

Angew Chem Int Ed Engl. 2023 Apr 04. e202212636

Apolipoprotein E Recognizes Alzheimer's Disease Associated 3-O Sulfation of Heparan Sulfate.

Dylan James Mah, Yanan Zhu, Guowei Su, Jing Zhao, Ashely Canning, James Gibson, Xuehong Song, Eduardo Stancanelli, Yongmei Xu, Fuming Zhang, Robert J Linhardt, Jian Liu, Lianchun Wang, Chunyu Wang.

  Apolipoprotein E (ApoE)'s ε4 alle is the most important genetic risk factor for late onset Alzheimer's Disease (AD). Cell-surface heparan sulfate (HS) is a cofactor for ApoE/LRP1 interaction and the prion-like spread of tau pathology between cells. 3- O -sulfo (3- O -S) modification of HS has been linked to AD through its interaction with tau, and enhanced levels of 3- O -sulfated HS and 3- O -sulfotransferases in AD brains. In this study, we characterized ApoE/HS interactions in wildtype ApoE3, AD-linked ApoE4, and AD-protective ApoE2 and ApoE3-Christchurch. Glycan microarray and SPR assays revealed that all ApoE isoforms recognized 3- O -S. NMR titration localized ApoE/3- O -S binding to the vicinity of canonical HS binding motif. In cells, the knockout of HS3ST1-a major 3- O -sulfotransferase-reduced cell surface binding and uptake of ApoE. 3- O -S is thus recognized by both tau and ApoE, suggesting that the interplay between 3- O -sulfated HS, tau and ApoE isoforms may modulate AD risk.

Keywords:  3-O-Sulfation; Alzheimer's disease; Glycobiology; Heparan sulfate; apolipoprotein E

DOI:  https://doi.org/10.1002/anie.202212636
Chem Sci. 2023 Mar 29. 14(13): 3514-3522

Heparan sulfate glycomimetics via iterative assembly of "clickable" disaccharides.

Cangjie Yang, Yu Deng, Yang Wang, Chaoshuang Xia, Akul Y Mehta, Kelly J Baker, Anuj Samal, Putthipong Booneimsri, Chanthakarn Lertmaneedang, Seung Hwang, James P Flynn, Muqing Cao, Chao Liu, Alec C Zhu, Richard D Cummings, Cheng Lin, Udayan Mohanty, Jia Niu.

Heparan sulfate (HS) glycosaminoglycans are widely expressed on the mammalian cell surfaces and extracellular matrices and play important roles in a variety of cell functions. Studies on the structure-activity relationships of HS have long been hampered by the challenges in obtaining chemically defined HS structures with unique sulfation patterns. Here, we report a new approach to HS glycomimetics based on iterative assembly of clickable disaccharide building blocks that mimic the disaccharide repeating units of native HS. Variably sulfated clickable disaccharides were facilely assembled into a library of mass spec-sequenceable HS-mimetic oligomers with defined sulfation patterns by solution-phase iterative syntheses. Microarray and surface plasmon resonance (SPR) binding assays corroborated molecular dynamics (MD) simulations and confirmed that these HS-mimetic oligomers bind protein fibroblast growth factor 2 (FGF2) in a sulfation-dependent manner consistent with that of the native HS. This work established a general approach to HS glycomimetics that can potentially serve as alternatives to native HS in both fundamental research and disease models.

DOI: https://doi.org/10.1039/d3sc00260h
Carbohydr Polym. 2023 Jul 01. pii: S0144-8617(22)01393-5. [Epub ahead of print]311 120488

Molecular weight determination of low molecular weight hyaluronic acid and chondroitin sulfate by gel permeation chromatography.

Zhiqing Tian, Fan Jiang, Haolong Cong, Shuifang Zhu.

  Low molecular weight (LWM) hyaluronic acid (HA) and chondroitin sulfate (CS) have a wide range of applications. To determine their molecular weight (MW), we developed a gel permeation chromatography (GPC) method, which is calibrated based on serrated peaks in the chromatograms. MW calibrants were obtained from the enzymolysis of HA and CS using hyaluronidase. The identical structure of calibrants and samples ensured the soundness of the method. The highest confidence MWs were up to 14,454 and 14,605 for HA and CS, respectively, and the standard curves showed very high correlation coefficients. Thanks to the changeless relationship between MW and its contribution to the GPC integral, the second calibration curves could be derived via one GPC column, also embodied correlation coefficients of >0.9999. The discrepancies of MW values were minuscule, and the measurement of a sample could be conducted in <30 min. The accuracy of the method was verified using LWM heparins, and the measured Mw values showed a 1.2 %-2.0 % error relative to pharmacopeia results. The MW results obtained for LWM-HA and LWM-CS samples were also consistent with the results obtained by multiangle laser light scattering. The method was also verified be able to measure the very low MWs.

Keywords:  Chondroitin sulfate; GPC; Hyaluronic acid; Low molecular weight; Molecular weight determination

DOI:  https://doi.org/10.1016/j.carbpol.2022.120488
Nutr Metab (Lond). 2023 Apr 04. 20(1): 22

Combined treatment with glucosamine and chondroitin sulfate improves rheumatoid arthritis in rats by regulating the gut microbiota.

Xuesong Wang, Dongsong Liu, Dan Li, Jiai Yan, Ju Yang, Xiaohui Zhong, Qin Xu, Yuanze Xu, Yanping Xia, Qinyue Wang, Hong Cao, Feng Zhang.

   BACKGROUND: To investigate the ameliorative effects of glucosamine (GS), chondroitin sulphate (CS) and glucosamine plus chondroitin sulphate (GC) on rheumatoid arthritis (RA) in rats, and to explore the mechanism of GS, CS and GC in improving RA based on the gut microbiota.
METHODS: RA rat models were effectively developed 14 days after CFA injection, and then garaged with GS, CS and GC. Body weight and paw volume of rats were monitored at multiple time points at the beginning of CFA injection. Until D36, serum and ankle tissue specimens were used to measure levels of circulating inflammatory factors (TNF-α, IL-1β, MMP-3, NO and PGE2) and local inflammatory indicators (TLR-4 and NF-κB). On D18, D25, and D36, intergroup gut microbiota was compared using 16S rRNA gene sequencing and bioinformatics analysis. We also performed the correlation analysis of gut bacteria, joint swelling and inflammatory indicators.
RESULTS: GC, rather than GS and CS, could reduce right paw volumes, levels of TLR-4 and NF-κB in synovial tissues. In addition, enriched genera in RA model rats screened out by LEfSe analysis could be inhibited by GC intervention, including potential LPS-producing bacteria (Enterobacter, Bacteroides, Erysipelotrichaceae_unclassified and Erysipelotrichaceae_uncultured) and some other opportunistic pathogens (Esherichia_Shigella, Nosocomiicoccus, NK4A214_group, Odoribacter, Corynebacterium and Candidatus_Saccharimonas.etc.) that positively correlated with pro-inflammatory cytokines, right paw volume, and pathology scores. Furthermore, the gut microbiota dysbiosis was observed to recover before alleviating joint swelling after interventions.
CONCLUSIONS: GC could inhibit potential LPS-producing bacteria and the activation of TLR-4/NF-κB pathway in RA rats, thus alleviating RA-induced joint injury.

Keywords:  Chondroitin sulfate; Glucosamine; Gut microbiota; Rheumatoid arthritis

DOI:  https://doi.org/10.1186/s12986-023-00735-2
Carbohydr Polym. 2023 Jul 01. pii: S0144-8617(23)00243-6. [Epub ahead of print]311 120779

Absolute pharmacokinetics of heparin in primates.

Yuefan Song, Ahmed Kouta, Lee M Cera, Ke Xia, Fuming Zhang, Roland Kraemer, Jawed Fareed, Robert J Linhardt, Walter Jeske.

  Heparin is a commonly used anticoagulant drug, derived from the tissues of animals including pigs, cows, and sheep. Measuring heparin concentration in plasma is challenging due to its complex molecular structure. Existing methods rely on measuring heparin's anticoagulant activity, which provides pharmacodynamic (PD) data but not pharmacokinetic (PK) data, measuring concentration over time. To overcome this limitation, we used liquid chromatography-mass spectrometry (LC-MS) and the multiple reaction monitoring (MRM) method to directly measure heparin's concentration in non-human primates after administering porcine, bovine, and ovine heparin. A protocol was developed to enable an MRM method for application to small plasma volumes without purification. The PK data obtained from LC-MS are then compared with the data obtained using the Heparin Red assay and the PD data determined using biochemical clinical assays. Results showed that LC-MS and Heparin Red assay measurements closely correlated with unfractionated heparin's biological activities, supporting the use of mass spectra and dye-binding assays to determine heparin levels in plasma. This study builds a way for the measurement of heparin concentration in plasma, which could lead to an improved understanding of heparin's metabolism and dosing safety.

Keywords:  Heparin; LCMS; LMWH; Pharmacodynamics; Pharmacokinetics; Primates

DOI:  https://doi.org/10.1016/j.carbpol.2023.120779
Front Immunol. 2023 ;14 1131146

Cholesterol sulfate limits neutrophil recruitment and gut inflammation during mucosal injury.

Kenji Morino, Kazufumi Kunimura, Yuki Sugiura, Yoshihiro Izumi, Keisuke Matsubara, Sayaka Akiyoshi, Rae Maeda, Kenichiro Hirotani, Daiji Sakata, Seiya Mizuno, Satoru Takahashi, Takeshi Bamba, Takehito Uruno, Yoshinori Fukui.

  During mucosal injury, intestinal immune cells play a crucial role in eliminating invading bacteria. However, as the excessive accumulation of immune cells promotes inflammation and delays tissue repair, it is essential to identify the mechanism that limits the infiltration of immune cells to the mucosal-luminal interface. Cholesterol sulfate (CS) is the lipid product of the sulfotransferase SULT2B1 and suppresses immune reactions by inhibiting DOCK2-mediated Rac activation. In this study, we aimed to elucidate the physiological role of CS in the intestinal tract. We found that, in the small intestine and colon, CS is predominantly produced in the epithelial cells close to the lumen. While dextran sodium sulfate (DSS)-induced colitis was exacerbated in Sult2b1-deficient mice with increased prevalence of neutrophils, the elimination of either neutrophils or intestinal bacteria in Sult2b1-deficient mice attenuated disease development. Similar results were obtained when the Dock2 was genetically deleted in Sult2b1-deficient mice. In addition, we also show that indomethacin-induced ulcer formation in the small intestine was exacerbated in Sult2b1-deficient mice and was ameliorated by CS administration. Thus, our results uncover that CS acts on inflammatory neutrophils, and prevents excessive gut inflammation by inhibiting the Rac activator DOCK2. The administration of CS may be a novel therapeutic strategy for inflammatory bowel disease and non-steroidal anti-inflammatory drug-induced ulcers.

Keywords:  CyTOF; DOCK2; SULT2B1; cholesterol sulfate; gut inflammation; mass spectrometry; neutrophil

DOI:  https://doi.org/10.3389/fimmu.2023.1131146
Anal Chim Acta. 2023 May 08. pii: S0003-2670(23)00349-5. [Epub ahead of print]1254 341128

Effects of structural characteristics of (un)conjugated steroid metabolites in their collision cross section value.

Claudia Bressan, Alberto Celma, Élida Alechaga, Nuria Monfort, Rosa Ventura, Juan Vicente Sancho.

  In this work, the collision cross section (CCS) value of 103 steroids (including unconjugated metabolites and phase II metabolites conjugated with sulfate and glucuronide groups) was determined by liquid chromatography coupled to traveling wave ion mobility spectrometry (LC-TWIMS). A time of flight (QTOF) mass analyzer was used to perform the analytes determination at high-resolution mass spectrometry. An electrospray ionization source (ESI) was used to generate [M+H]+, [M + NH4]+ and/or [M - H]- ions. High reproducibility was observed for the CCS determination in both urine and standard solutions, obtaining RSD lower than 0.3% and 0.5% in all cases respectively. CCS determination in matrix was in accordance with the CCS measured in standards solution showing deviations below 2%. In general, CCS values were directly correlated with the ion mass and allowed differentiating between glucuronides, sulfates and free steroids although differences among steroids of the same group were less significant. However, more specific information was obtained for phase II metabolites observing differences in the CCS value of isomeric pairs concerning the conjugation position or the α/β configuration, which could be useful in the structural elucidation of new steroid metabolites in the anti-doping field. Finally, the potential of IMS reducing interferences from the sample matrix was also tested for the analysis of a glucuronide metabolite of bolasterone (5β-androstan-7α,17α-dimethyl-3α,17β-diol-3-glucuronide) in urine samples.

Keywords:  Anti-doping; Collision cross section; Ion mobility; Isomeric compounds; Urine analysis

DOI:  https://doi.org/10.1016/j.aca.2023.341128
Plant Sci. 2023 Apr 04. pii: S0168-9452(23)00114-0. [Epub ahead of print] 111697

Nitric oxide and hydrogen peroxide mediated regulation of chromium (VI) toxicity in wheat seedlings involves alterations in antioxidants and high affinity sulfate transporter.

Samiksha Singh, Nawal Kishore Dubey, Durgesh Kumar Tripathi, Ravi Gupta, Vijay Pratap Singh.

  Chromium contamination of the soil is a major scientific concern with reference to crop productivity and human health. In recent years, several approaches are being employed in managing metal toxicity in crop plants. Here, we have investigated about potential and probable crosstalk of nitric oxide (NO) and hydrogen peroxide (H2O2) in mitigating hexavalent chromium [Cr(VI)] toxicity in wheat seedlings. Cr(VI) toxicity reduced the fresh mass and overall growth due to accumulation of reactive oxygen species (ROS) and decreased efficiency of AsA-GSH cycle and downregulation of high affinity sulfate transporter. However, exogenous treatment of NO and H2O2 significantly alleviated Cr toxicity. Application of NO and ROS scavengers reversed stress mitigating effects of NO and H2O2, respectively suggesting that endogenous NO and H2O2 are necessary for rendering Cr toxicity tolerance. Furthermore, NO rescued negative effect of diphenylene iodonium (DPI, NADPH oxidase inhibitor) and H2O2 reversed the negative effect of c-PTIO suggesting that they exhibit independent signalling in mitigating Cr stress. Altogether, data indicated that NO and H2O2 rendered mitigation of Cr stress by up-regulating enzymes (activity and relative gene expression) and metabolites of ascorbate-glutahione cycle, high affinity sulfate transporter (relative gene expression) and glutathione biosynthesis which collectively controlled occurrence of oxidative stress.

Keywords:  Ascorbate-glutathione cycle; Glutathione biosynthesis; Growth; High affinity sulfate transporter; Oxidative stress indices, Relative gene expression

DOI:  https://doi.org/10.1016/j.plantsci.2023.111697
JACS Au. 2023 Mar 27. 3(3): 628-656

Glycosaminoglycans: What Remains To Be Deciphered?

Serge Perez, Olga Makshakova, Jesus Angulo, Emiliano Bedini, Antonella Bisio, Jose Luis de Paz, Elisa Fadda, Marco Guerrini, Michal Hricovini, Milos Hricovini, Frederique Lisacek, Pedro M Nieto, Kevin Pagel, Giulia Paiardi, Ralf Richter, Sergey A Samsonov, Romain R Vivès, Dragana Nikitovic, Sylvie Ricard Blum.

Glycosaminoglycans (GAGs) are complex polysaccharides exhibiting a vast structural diversity and fulfilling various functions mediated by thousands of interactions in the extracellular matrix, at the cell surface, and within the cells where they have been detected in the nucleus. It is known that the chemical groups attached to GAGs and GAG conformations comprise "glycocodes" that are not yet fully deciphered. The molecular context also matters for GAG structures and functions, and the influence of the structure and functions of the proteoglycan core proteins on sulfated GAGs and vice versa warrants further investigation. The lack of dedicated bioinformatic tools for mining GAG data sets contributes to a partial characterization of the structural and functional landscape and interactions of GAGs. These pending issues will benefit from the development of new approaches reviewed here, namely (i) the synthesis of GAG oligosaccharides to build large and diverse GAG libraries, (ii) GAG analysis and sequencing by mass spectrometry (e.g., ion mobility-mass spectrometry), gas-phase infrared spectroscopy, recognition tunnelling nanopores, and molecular modeling to identify bioactive GAG sequences, biophysical methods to investigate binding interfaces, and to expand our knowledge and understanding of glycocodes governing GAG molecular recognition, and (iii) artificial intelligence for in-depth investigation of GAGomic data sets and their integration with proteomics.

DOI: https://doi.org/10.1021/jacsau.2c00569