bims-supasi 2022-08-28 papers

bims-supasi

Biomed News

on Sulfation pathways and signalling

Issue of 2022–08–28
twelve papers selected by
Jonathan Wolf Mueller, University of Birmingham

Cellular Chondroitin Sulfate and the Mucin-like Domain of Viral Glycoprotein C Promote Diffusion of Herpes Simplex Virus 1 While Heparan Sulfate Restricts Mobility.
Sulfated modification of hyaluronan tetrasaccharide enhances its antitumor activity on human lung adenocarcinoma A549 cells in vitro and in vivo.
Alterations in glycosaminoglycan biosynthesis associated with the Ehlers-Danlos Syndromes.
Enzymatic synthesis of low molecular weight heparins from N-sulfo heparosan depolymerized by heparanase or heparin lyase.
The Role of the Glypican and Syndecan Families of Heparan Sulfate Proteoglycans in Cardiovascular Function and Disease.
Heparan Sulfate Facilitates Binding of hIFNγ to Its Cell-Surface Receptor hIFNGR1.
Synthesis, Chemical and Biomedical Aspects of the Use of Sulfated Chitosan.
Cryostructuring of Polymeric Systems: 62 Preparation and Characterization of Alginate/Chondroitin Sulfate Cryostructurates Loaded with Antimicrobial Substances.
SARS-CoV-2 triggers complement activation through interactions with heparan sulfate.
Heparin: An old drug for new clinical applications.
Transcription level and phylogeny analyses of Chlamydomonas reinhardtii arylsulfatases.
A 'new' estrogen metabolite: an epoxide of estrone as a sulfated steroid.

Viruses. 2022 Aug 21. pii: 1836. [Epub ahead of print]14(8):

Cellular Chondroitin Sulfate and the Mucin-like Domain of Viral Glycoprotein C Promote Diffusion of Herpes Simplex Virus 1 While Heparan Sulfate Restricts Mobility.

Yara Abidine, Lifeng Liu, Oskar Wallén, Edward Trybala, Sigvard Olofsson, Tomas Bergström, Marta Bally.

  The diffusion of viruses at the cell membrane is essential to reach a suitable entry site and initiate subsequent internalization. Although many viruses take advantage of glycosaminoglycans (GAG) to bind to the cell surface, little is known about the dynamics of the virus-GAG interactions. Here, single-particle tracking of the initial interaction of individual herpes simplex virus 1 (HSV-1) virions reveals a heterogeneous diffusive behavior, regulated by cell-surface GAGs with two main diffusion types: confined and normal free. This study reports that different GAGs can have competing influences in mediating diffusion on the cells used here: chondroitin sulfate (CS) enhances free diffusion but hinders virus attachment to cell surfaces, while heparan sulfate (HS) promotes virus confinement and increases entry efficiency. In addition, the role that the viral mucin-like domains (MLD) of the HSV-1 glycoprotein C plays in facilitating the diffusion of the virus and accelerating virus penetration into cells is demonstrated. Together, our results shed new light on the mechanisms of GAG-regulated virus diffusion at the cell surface for optimal internalization. These findings may be extendable to other GAG-binding viruses.

Keywords:  glycocalyx; glycosaminoglycan; herpesvirus; mucin-like domain; single particle tracking; viral O-glycans; virus diffusion

DOI:  https://doi.org/10.3390/v14081836
Bioorg Med Chem Lett. 2022 Aug 17. pii: S0960-894X(22)00421-8. [Epub ahead of print] 128945

Sulfated modification of hyaluronan tetrasaccharide enhances its antitumor activity on human lung adenocarcinoma A549 cells in vitro and in vivo.

Xinyue He, Sanying Wang, Bing Liu, Di Jiang, Fen Chen, Genxiang Mao, Weihua Jin, Hongying Pan, Weihong Zhong.

  Hyaluronan (HA) is a glycosaminoglycan polymer involved in cell phenotype change, inflammation modulation, and tumor metastasis progression. HA oligosaccharides have a higher solubility and drug-forming ability than polysaccharides. HA tetrasaccharide was reported as the smallest fragment required for inhibiting triple-negative breast cancer, but the anti-tumor activity of HA tetrasaccharide (HA4) and its sulfated derivatives in lung cancer is still unknown. In this study, HA4 was prepared via HA degradation by chondroitin sulfate ABC (CSABC), while its sulfated derivatives were prepared by sulfur pyridine trioxide complex in N, N-dimethylformamide (DMF). Then, the anti-tumor activity was detected via MTT assay and xenograft tumor experiments, while the expression level change of apoptosis genes was analyzed by qRT-PCR. Electrospray mass spectrometry (ESI-MS) analysis showed several HA4 sulfated derivatives, GlcA2GlcNAc2 (SO3H)n contains 0-6 sulfation groups, which mainly contain 3-6, 2-3, and 0-1 sulfation groups were classified as HA4S1, HA4S2, and HA4S3, respectively. After the addition of 1.82 mg/mL HA4, HA4S1, HA4S2, and HA4S3, the cell viability of A549 cells was reduced to 81.2%, 62.1%, 50.3%, and 65.9%, respectively. Thus, HA4S2 was chosen for further measurement, the qRT-PCR results showed it significantly up-regulated the expression of genes in the apoptosis pathway. Moreover, HA4S2 exhibited stronger antitumor activity than HA4 in vivo and the tumor inhibition rate reached 36.90%. In summary, this study indicated that the CSABC enzyme could effectively degrade HA into oligosaccharides, and sulfation modification was an effective method to enhance the antitumor activity of HA tetrasaccharides.

Keywords:  Hyaluronan; antitumor; apoptosis; sulfation modification; tetrasaccharide

DOI:  https://doi.org/10.1016/j.bmcl.2022.128945
Am J Physiol Cell Physiol. 2022 Aug 22.

Alterations in glycosaminoglycan biosynthesis associated with the Ehlers-Danlos Syndromes.

Delfien Syx, Sarah Delbaere, Catherine Bui, Adelbert De Clercq, Göran Larson, Shuji Mizumoto, Tomoki Kosho, Sylvie Fournel-Gigleux, Fransiska Malfait.

  Proteoglycans consist of a core protein substituted with one or more glycosaminoglycan (GAG) chains and execute versatile functions during many physiological and pathological processes. The biosynthesis of GAG chains is a complex process that depends on the concerted action of a variety of enzymes. Central to the biosynthesis of heparan sulfate (HS) and chondroitin sulfate/dermatan sulfate (CS/DS) GAG chains is the formation of a tetrasaccharide linker region followed by biosynthesis of HS or CS/DS-specific repeating disaccharide units, which then undergo modifications and epimerisation. The importance of these biosynthetic enzymes is illustrated by several severe pleiotropic disorders that arise upon their deficiency. The Ehlers-Danlos syndrome (EDS) constitute a special group among these disorders. While most EDS types are caused by defects in fibrillar types I, III or V collagen, or their modifying enzymes, a few rare EDS types have recently been linked to defects in GAG biosynthesis. Spondylodysplastic EDS (spEDS) is caused by defective formation of the tetrasaccharide linker region, either due to β4GalT7 or β3GalT6 deficiency, whereas musculocontractural EDS (mcEDS) results from deficiency of D4ST1 or DS-epi1, impairing DS formation. This narrative review highlights the consequences of GAG deficiency in these specific EDS types, summarizes the associated phenotypic features and the molecular spectrum of reported pathogenic variants and defines the current knowledge on the underlying pathophysiological mechanisms based on studies in patient-derived material, in vitro analyses and animal models.

Keywords:  EDS; Ehlers-Danlos syndromes; collagen; glycosaminoglycan; proteoglycan

DOI:  https://doi.org/10.1152/ajpcell.00127.2022
Carbohydr Polym. 2022 Nov 01. pii: S0144-8617(22)00730-5. [Epub ahead of print]295 119825

Enzymatic synthesis of low molecular weight heparins from N-sulfo heparosan depolymerized by heparanase or heparin lyase.

Yanlei Yu, Li Fu, Peng He, Ke Xia, Sony Varghese, Jonathan Dordick, Hong Wang, Fuming Zhang, Robert J Linhardt.

  Low-molecular-weight heparin (LMWH) is prepared from the controlled chemical or enzymatic depolymerization of animal sourced heparins. It has been widely used as an anticoagulant. Concerns about the shortcomings of animal-derived heparin and the contamination of supply chain demand biochemical approaches for synthesizing LMWH. In the present study, two LMWHs were enzymatically synthesized from low molecular weight N-sulfated heparosan (LMW-NSH) cleaved by recombinant hydrolase, endo-β-glucuronidase, (HepBp) or heparin lyase III (HepIII), followed by subsequent sulfotransferase modifications. Structural characterization shows that LMWH chains prepared using HepBp had a saturated uronic acid residue at their reducing ends, while chains of LMWH prepared using HepIII had an unsaturated uronic acid residue at their non-reducing end. Both LMWHs had anti-factor Xa and anti-factor IIa activities comparable to enoxaparin. This approach demonstrates that the hydrolase, HepBp, can be used to prepare a new type of LMWH that has no unsaturated uronic acid at its non-reducing end.

Keywords:  Anticoagulant activity; Enzymatic synthesis; Heparanase Bp; Heparinase III; Low molecular weight heparin

DOI:  https://doi.org/10.1016/j.carbpol.2022.119825
Am J Physiol Cell Physiol. 2022 Aug 22.

The Role of the Glypican and Syndecan Families of Heparan Sulfate Proteoglycans in Cardiovascular Function and Disease.

Lakshmi Narasimha Rao Thota, Andreia Zago Chignalia.

  Heparan sulfate proteoglycans (HSPGs) are proteoglycans formed by a core protein to which one or multiple heparan sulfate chains are covalently bound. They are ubiquitously expressed in cellular surfaces and can be found in the extracellular matrix and secretory vesicles. The cellular effects of HSPGs comprehend multiple functionalities that include: (i) the interaction with other membrane surface proteins to act as a substrate for cellular migration; (ii) acting as a binding site for circulating molecules; (iii) to have a receptor role for proteases; (iv) to act as a coreceptor that can provide finetuning of growth factor receptor activity threshold; and (v) to activate intracellular signaling pathways [1]. Among the different families of HSPGs, the syndecan and glypican families of HSPGs have gained increased attention in relation to their effects on cardiovascular cells and potential role in disease progression. In this mini-review we will summarize the effects of syndecan and glypican homologs on the different cardiovascular cell types and discuss their contribution to common processes found in cardiovascular diseases (inflammation, hypertrophy, and vascular remodeling) as well as their potential role in the development and progression of specific diseases including hypertension, heart failure, and atherosclerosis.

Keywords:  HSPGs; cardiovascular; glypican; syndecan

DOI:  https://doi.org/10.1152/ajpcell.00018.2022
Int J Mol Sci. 2022 Aug 20. pii: 9415. [Epub ahead of print]23(16):

Heparan Sulfate Facilitates Binding of hIFNγ to Its Cell-Surface Receptor hIFNGR1.

Elisaveta Miladinova, Elena Lilkova, Elena Krachmarova, Kristina Malinova, Peicho Petkov, Nevena Ilieva, Genoveva Nacheva, Leandar Litov.

  Human interferon-gamma (hIFNγ) is a crucial signaling molecule with an important role in the initialization and development of the immune response of the host. However, its aberrant activity is also associated with the progression of a multitude of autoimmune and other diseases, which determines the need for effective inhibitors of its activity. The development of such treatments requires proper understanding of the interaction of hIFNγ to its cell-surface receptor hIFNGR1. Currently, there is no comprehensive model of the mechanism of this binding process. Here, we employ molecular dynamics simulations to study on a microscopic level the process of hIFNγ-hIFNGR1 complex formation in different scenarios. We find that the two molecules alone fail to form a stable complex, but the presence of heparan-sulfate-like oligosaccharides largely facilitates the process by both demobilizing the highly flexible C-termini of the cytokine and assisting in the proper positioning of its globule between the receptor subunits. An antiproliferative-activity assay on cells depleted from cell-surface heparan sulfate (HS) sulfation together with the phosphorylation levels of the signal transducer and activator of transcription STAT1 confirms qualitatively the simulation-based multistage complex-formation model. Our results reveal the key role of HS and its proteoglycans in all processes involving hIFNγ signalling.

Keywords:  heparan sulfate; human interferon gamma; human interferon gamma receptor; molecular dynamics simulations; sodium chlorate

DOI:  https://doi.org/10.3390/ijms23169415
Polymers (Basel). 2022 Aug 22. pii: 3431. [Epub ahead of print]14(16):

Synthesis, Chemical and Biomedical Aspects of the Use of Sulfated Chitosan.

I N Bolshakov, L M Gornostaev, O I Fominykh, A V Svetlakov.

  This work is devoted to the chemical synthesis of sulfated chitosan and its experimental verification in an animal model of early atherosclerosis. The method of chitosan quaternization with sulfate-containing ingredients resulted in a product with a high content of sulfate groups. Implantation of this product into the fascial-muscular sheath of the main limb artery along the leg and thigh in rabbits led to the extraction of cholesterol from the subintimal region. Simplified methods for the chemical synthesis of quaternized sulfated chitosan and the use of these products in a model of experimental atherosclerosis made it possible to perform a comparative morphological analysis of the vascular walls of the experimental and control limbs under conditions of a long-term high-cholesterol diet. The sulfated chitosan samples after implantation were shown to change the morphological pattern of the intimal and middle membranes of the experimental limb artery. The implantation led to the degradation of soft plaques within 30 days after surgical intervention, which significantly increased collateral blood flow. The implantation of sulfated chitosan into the local area of the atherosclerotic lesions in the artery can regulate the cholesterol content in the vascular wall and destroy soft plaques in the subintimal region.

Keywords:  O,N-(2-sulfoethyl)chitosan; experimental atherosclerosis; organic synthesis; para-adventitial implantation; sulfated polyelectrolyte complexes; transport system; vascular cholesterol extraction

DOI:  https://doi.org/10.3390/polym14163431
Polymers (Basel). 2022 Aug 11. pii: 3271. [Epub ahead of print]14(16):

Cryostructuring of Polymeric Systems: 62 Preparation and Characterization of Alginate/Chondroitin Sulfate Cryostructurates Loaded with Antimicrobial Substances.

Olga I Vernaya, Andrey N Ryabev, Tatyana I Shabatina, Daria L Karlova, Andrey V Shabatin, Lyudmila N Bulatnikova, Alexander M Semenov, Mikhail Ya Melnikov, Vladimir I Lozinsky.

  Targeted drug release is a significant research focus in the development of drug delivery systems and involves a biocompatible polymeric carrier and certain medicines. Cryostructuring is a suitable approach for the preparation of efficient macroporous carriers for such drug delivery systems. In the current study, the cryogenically structured carriers based on alginate/chondroitin sulfate mixtures were prepared and their physicochemical properties and their ability to absorb/release the bactericides were evaluated. The swelling parameters of the polysaccharide matrix, the amount of the tightly bound water in the polymer and the sulfur content were measured. In addition, FTIR and UV spectroscopy, optical and scanning microscopy, as well as a standard disk diffusion method for determining antibacterial activity were used. It was shown that alginate/chondroitin sulfate concentration and their ratios were significant factors influencing the swelling properties and the porosity of the resultant cryostructurates. It was demonstrated that the presence of chondroitin sulfate in the composition of a polymeric matrix slowed down the release of the aminoglycoside antibiotic gentamicin. In the case of the NH2-free bactericide, dioxidine, the release was almost independent of the presence of chondroitin sulfate. This trend was also registered for the antibacterial activity tests against the Escherichia coli bacteria, when examining the drug-loaded biopolymeric carriers.

Keywords:  alginate; chondroitin sulfate; controlled drug release systems; cryostructurates; dioxidine; gentamicin sulfate

DOI:  https://doi.org/10.3390/polym14163271
Clin Transl Immunology. 2022 ;11(8): e1413

SARS-CoV-2 triggers complement activation through interactions with heparan sulfate.

Martin W Lo, Alberto A Amarilla, John D Lee, Eduardo A Albornoz, Naphak Modhiran, Richard J Clark, Vito Ferro, Mohit Chhabra, Alexander A Khromykh, Daniel Watterson, Trent M Woodruff.

   Objectives: To determine whether SARS-CoV-2 can trigger complement activation, the pathways that are involved and the functional significance of the resultant effect.
Methods: SARS-CoV-2 was inoculated into a human lepirudin-anticoagulated whole blood model, which contains a full repertoire of complement factors and leukocytes that express complement receptors. Complement activation was determined by measuring C5a production with an ELISA, and pretreatment with specific inhibitors was used to identify the pathways involved. The functional significance of this was then assessed by measuring markers of C5a signalling including leukocyte C5aR1 internalisation and CD11b upregulation with flow cytometry.
Results: SARS-CoV-2 inoculation in this whole blood model caused progressive C5a production over 24 h, which was significantly reduced by inhibitors for factor B, C3, C5 and heparan sulfate. However, this phenomenon could not be replicated in cell-free plasma, highlighting the requirement for cell surface interactions with heparan sulfate. Functional analysis of this phenomenon revealed that C5aR1 signalling and CD11b upregulation in granulocytes and monocytes was delayed and only occurred after 24 h.
Conclusion: SARS-CoV-2 is a noncanonical alternative pathway activator that progressively triggers complement activation through interactions with heparan sulfate.

Keywords:  SARS‐CoV‐2; alternative pathway; complement; coronavirus; heparan sulfate; leukocyte activation

DOI:  https://doi.org/10.1002/cti2.1413
Carbohydr Polym. 2022 Nov 01. pii: S0144-8617(22)00723-8. [Epub ahead of print]295 119818

Heparin: An old drug for new clinical applications.

Peipei Wang, Lianli Chi, Zhenqing Zhang, Hongmei Zhao, Fuming Zhang, Robert J Linhardt.

  Heparin, an old but first-line anticoagulant, has been used over a century. It is a heterogeneous, linear, highly sulfated, anionic glycosaminoglycan with a broad distribution in relative molecular weight and charge density. These structural properties allow heparin to selectively interact with multiple proteins, leading to heparin's various pharmacological functions, such as anticoagulant, anti-viral, anti-tumor and anti-inflammatory activities. Clinical data suggest that unfractionated heparin or low molecule weight heparin could decrease mortality in COVID-19 patients with sepsis-induced hypercoagulation through the anticoagulant, anti-viral and anti-inflammatory activities of these drugs. Thus, the non-anticoagulant activity of heparin has again aroused attention. This review highlights recent advances in the preparation of heparin-derived drugs and clinical research on its non-anticoagulant properties over the past decade, to further the development and utilization of these important drugs.

Keywords:  Clinical application; Heparin; Non-anticoagulant bioactivity; Preparation

DOI:  https://doi.org/10.1016/j.carbpol.2022.119818
J Eukaryot Microbiol. 2022 Aug 26. e12943

Transcription level and phylogeny analyses of Chlamydomonas reinhardtii arylsulfatases.

Fatma Salarvan, Hasan Meydan, Münevver Aksoy.

  Sulfur is a required macroelement for all organisms and sulfate deficiency causes growth and developmental defects. Arylsulfatases (ARS) hydrolyze sulfate from sulfate esters and make sulfate bioavailable for plant uptake. These enzymes are found in microorganisms and animals, however plant genomes do not encode any ARS gene. Our database searches found nineteen ARS genes in the genome of Chlamydomonas reinhardtii. Among these, ARS1 and ARS2 were studied in the literature; however, the remaining seventeen gene models were not studied. Our results show that putative polypeptide sequences of the ARS gene models all have the sulfatase domain and sulfatase motifs found in known arylsulfatases. Phylogenetic analyses show that C. reinhardtii proteins are in close branches with Volvox carterii proteins while they were clustered in a separate group from Homo sapiens and bacterial species (Pseudomonas aeruginosa and Rhodopirellula baltica SH1), except human Sulf1, Sulf2 and GNS are clustered with algal ARSs. RT-PCR analyses showed that transcription of ARS6, ARS7, ARS11, ARS12, ARS13, ARS17 and ARS19 increased under sulfate deficiency. However, this increase was not as high as the increase seen in ARS2. Since plant genomes do not encode any ARS gene, our results highlight the importance of microbial ARS genes.

Keywords:  Ars gene; gene duplication; phylogeny; sulfate; sulfate deficiency

DOI:  https://doi.org/10.1111/jeu.12943
J Endocrinol. 2022 Aug 01. pii: JOE-22-0177. [Epub ahead of print]

A 'new' estrogen metabolite: an epoxide of estrone as a sulfated steroid.

James I Raeside, Heather L Christie.

Current heightened recognition of the importance of sulfated steroids led to examination of conjugates in media from incubations of estrogens in tissues from the reproductive tract of stallions. Previously we had reported a 'new' unidentified metabolite of estrone (E1) and [3H]-E1, located between 17β-estradiol (E2) and E1 reference standards on chromatography (HPLC), and identified tentatively as a stable 5α,6α-estrone epoxide. Stallion tissues were minced and incubated for 2h with [3H]-E1 (1x106 cpm). Solid-phase extraction of unconjugated and conjugated steroids from media was followed by liquid scintillation counting (LSC), where radioactivity was mostly in the conjugate fractions (>80%). HPLC of conjugated steroids used an isocratic solvent system of acetonitrile/water (8:92) at 700 µl/min with detection by LSC. A radioactive peak between reference standards of E1 and E2 sulfates was examined, after solvolysis, in a second solvent system. Sulfated steroids yielded largely E1, whereas acid treatment of the unconjugated E1 epoxide had earlier formed 6α-OH-E1 almost exclusively. With sulfatase enzyme, at neutral pH, radioactivity was also seen mostly as E1 with trace amounts of polar material. Reduction with KBH4, however, led also to desulfation; radioactivity had alignment with E2, but even more had low retention times as for 6α/6β-OH-E2. These findings point to a different hydrolysis for desulfation; even more, they reveal an additional oxygen atom at C6 and are supportive of biological formation of 5α,6α-epoxides of E1 and E2. As possible alternatives to catechol estrogens, implicated in cancer, the 'new' estrogen metabolites and their sulfated forms may have special interest.

DOI: https://doi.org/10.1530/JOE-22-0177