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Li T, Spruit CM, Wei N, Liu L, Wolfert MA, de Vries RP, Boons GJ. Chemoenzymatic Synthesis of Tri-antennary N-Glycans Terminating in Sialyl-Lewis x Reveals the Importance of Glycan Complexity for Influenza A Virus Receptor Binding. Chemistry 2024; 30:e202401108. [PMID: 38567703 PMCID: PMC11156558 DOI: 10.1002/chem.202401108] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Grants] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 03/25/2024] [Indexed: 05/09/2024]
Abstract
Sialyl-Lewisx (SLex) is involved in immune regulation, human fertilization, cancer, and bacterial and viral diseases. The influence of the complex glycan structures, which can present SLex epitopes, on binding is largely unknown. We report here a chemoenzymatic strategy for the preparation of a panel of twenty-two isomeric asymmetrical tri-antennary N-glycans presenting SLex-Lex epitopes on either the MGAT4 or MGAT5 arm that include putative high-affinity ligands for E-selectin. The N-glycans were prepared starting from a sialoglycopeptide isolated from egg yolk powder and took advantage of inherent substrate preferences of glycosyltransferases and the use of 5'-diphospho-N-trifluoracetylglucosamine (UDP-GlcNHTFA) that can be transferred by branching N-acetylglucosaminyltransferases to give, after base treatment, GlcNH2-containing glycans that temporarily disable an antenna from enzymatic modification. Glycan microarray binding studies showed that E-selectin bound equally well to linear glycans and tri-antennary N-glycans presenting SLex-Lex. On the other hand, it was found that hemagglutinins (HA) of H5 influenza A viruses (IAV) preferentially bound the tri-antennary N-glycans. Furthermore, several H5 HAs preferentially bound to N-glycan presenting SLex on the MGAT4 arm. SLex is displayed in the respiratory tract of several avian species, demonstrating the relevance of investigating the binding of, among others IAVs, to complex N-glycans presenting SLex.
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Affiliation(s)
- Tiehai Li
- Complex Carbohydrate Research Center, University of Georgia, Athens, GA 30602, USA
- Present address: Shanghai Institute of Materia Medica, Chinese Academy of Sciences, Shanghai, P. R. China
| | - Cindy M Spruit
- Department of Chemical Biology and Drug Discovery, Utrecht Institute for Pharmaceutical Sciences, Utrecht University, 3584 CG, Utrecht, The Netherlands
| | - Na Wei
- Complex Carbohydrate Research Center, University of Georgia, Athens, GA 30602, USA
| | - Lin Liu
- Complex Carbohydrate Research Center, University of Georgia, Athens, GA 30602, USA
| | - Margreet A Wolfert
- Complex Carbohydrate Research Center, University of Georgia, Athens, GA 30602, USA
- Department of Chemical Biology and Drug Discovery, Utrecht Institute for Pharmaceutical Sciences, Utrecht University, 3584 CG, Utrecht, The Netherlands
| | - Robert P de Vries
- Department of Chemical Biology and Drug Discovery, Utrecht Institute for Pharmaceutical Sciences, Utrecht University, 3584 CG, Utrecht, The Netherlands
| | - Geert-Jan Boons
- Complex Carbohydrate Research Center, University of Georgia, Athens, GA 30602, USA
- Department of Chemical Biology and Drug Discovery, Utrecht Institute for Pharmaceutical Sciences, Utrecht University, 3584 CG, Utrecht, The Netherlands
- Bijvoet Center for Biomolecular Research, Utrecht University, Utrecht, The Netherlands
- Chemistry Department, University of Georgia, Athens, GA 30602, USA
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Simpson HV, Umair S, Hoang VC, Savoian MS. Histochemical study of the effects on abomasal mucins of Haemonchus contortus or Teladorsagia circumcincta infection in lambs. Vet Parasitol 2016; 226:210-21. [PMID: 27387375 DOI: 10.1016/j.vetpar.2016.06.026] [Citation(s) in RCA: 7] [Impact Index Per Article: 0.8] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 05/23/2016] [Revised: 06/18/2016] [Accepted: 06/20/2016] [Indexed: 12/18/2022]
Abstract
Previously, chemical analysis of gastric fundic mucin showed that infection of sheep with Haemonchus contortus or Teladorsagia circumcincta changed the proportions of monosaccharides and decreased terminal mucin fucosylation and sialylation. To identify the effects of these parasites on the two mucin-secreting cell lineages, fundic and antral tissues were collected for histochemistry from 69 lambs aged from 3-4 to 9-10 months-of-age which had received a single infection of either H. contortus or T. circumcincta and euthanased at Day 21 or 28 post- infection respectively. All fundic tissues were stained separately with: (1) with Periodic Acid Schiff (PAS) for all mucins; (2) Alcian Blue (AB) pH 2.5 for acidic mucins (sialylated and sulphated); (3) AB pH 1 for sulphated mucins and (4) High Iron Diamine (HID) for sulphated mucins. Antral and fundic tissues from 24 lambs were also stained for acidic and neutral mucins or with specific lectins for α-1-linked fucose and for α-2,3- and α-2,6-linked sialic acids. Only mucin sulphation appeared to differ visually in uninfected lambs over this age range: there was weak staining with HID in tissues from lambs 3-6 months-of-age, but was generally more intense in those over 7 months-of-age. Sulphomucins were not apparent in surface mucous cells (SMC) or generally in the upper pits. Sialylomucins were located predominantly in the pits and glands, with small amounts of sialylated mucins in SMC and on the luminal surface, mainly in younger animals up to 6 months-of-age and less in the older animals. Parasitism markedly reduced the predominantly neutral surface mucin5AC of the SMC and pit cells, despite pit elongation in both antrum and fundus, whereas the acidic Muc6 secreted by mucus neck cells (MNC) increased along with MNC hyperplasia. Sulphated mucins were present mainly from the mid-pits downward and heavy staining was more common in older animals. In these sheep, the markedly reduced neutral mucin in the SMC and pit cells in both antrum and fundus contrasts with reported hypersecretion of mucus in the intestine, which is believed to aid in parasite expulsion. It has been proposed that intestinal goblet cell hypersecretion occurs only in resistant animals, therefore reduced mucins in the abomasum may be indicative of susceptibility to abomasal parasites.
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Affiliation(s)
- H V Simpson
- Institute of Veterinary, Animal and Biomedical Sciences, Massey University, Private Bag 11-222, Palmerston North, New Zealand.
| | - S Umair
- The Hopkirk Research Institute, AgResearch Ltd., Private Bag 11-008, Palmerston North, New Zealand
| | - V C Hoang
- Institute of Veterinary, Animal and Biomedical Sciences, Massey University, Private Bag 11-222, Palmerston North, New Zealand
| | - M S Savoian
- Institute of Fundamental Sciences, Massey University, Private Bag 11-222, Palmerston North, New Zealand
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Santra A, Yu H, Tasnima N, Muthana MM, Li Y, Zeng J, Kenyond NJ, Louie AY, Chen X. Systematic Chemoenzymatic Synthesis of O-Sulfated Sialyl Lewis x Antigens. Chem Sci 2016; 7:2827-2831. [PMID: 28138383 PMCID: PMC5269574 DOI: 10.1039/c5sc04104j] [Citation(s) in RCA: 32] [Impact Index Per Article: 3.6] [Reference Citation Analysis] [Abstract] [Grants] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 10/29/2015] [Accepted: 12/14/2015] [Indexed: 11/21/2022] Open
Abstract
O-Sulfated sialyl Lewis x antigens play important roles in nature. However, due to their structural complexity, they are not readily accessible by either chemical or enzymatic synthetic processes. Taking advantage of a bacterial sialyltransferase mutant that can catalyze the transfer of different sialic acid forms from the corresponding sugar nucleotide donors to Lewis x antigens which are fucosylated glycans as well as an efficient one-pot multienzyme (OPME) sialylation system, O-sulfated sialyl Lewis x antigens containing different sialic acid forms and O-sulfation at different locations were systematically synthesized by chemoenzymatic methods.
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Affiliation(s)
- Abhishek Santra
- Department of Chemistry, University of California, Davis One Shields Avenue, Davis, CA 95616 (USA)
| | - Hai Yu
- Department of Chemistry, University of California, Davis One Shields Avenue, Davis, CA 95616 (USA)
| | - Nova Tasnima
- Department of Chemistry, University of California, Davis One Shields Avenue, Davis, CA 95616 (USA)
| | - Musleh M Muthana
- Department of Chemistry, University of California, Davis One Shields Avenue, Davis, CA 95616 (USA)
| | - Yanhong Li
- Department of Chemistry, University of California, Davis One Shields Avenue, Davis, CA 95616 (USA)
| | - Jie Zeng
- Department of Chemistry, University of California, Davis One Shields Avenue, Davis, CA 95616 (USA) ; School of Food Science, Henan Institute of Science and Technology, Xinxiang, 453003 (China)
| | - Nicholas J Kenyond
- Division of Pulmonary, Critical Care and Sleep Medicine, Department of Internal Medicine, University of California, Davis, CA 95616 (USA)
| | - Angelique Y Louie
- Department of Biomedical Engineering, University of California, Davis, CA 95616 (USA)
| | - Xi Chen
- Department of Chemistry, University of California, Davis One Shields Avenue, Davis, CA 95616 (USA)
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Davis RA, Fettinger JC, Gervay-Hague J. Synthesis of cholesteryl-α-D-lactoside via generation and trapping of a stable β-lactosyl iodide. Tetrahedron Lett 2015; 56:3690-3694. [PMID: 26543257 DOI: 10.1016/j.tetlet.2015.05.012] [Citation(s) in RCA: 4] [Impact Index Per Article: 0.4] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/24/2022]
Abstract
The generation of β-lactosyl iodide was carried out under non-in situ-anomerization, metal free conditions by reacting commercially available β-per-O-acetylated lactose with trimethylsilyl iodide (TMSI). The β-iodide was surprisingly stable as evidenced by NMR spectroscopy. Introduction of octanol or cholesterol under microwave conditions gave high yields of α-linked glycoconjugates. Careful analysis of the reaction products and mechanistic considerations suggest an acid catalyzed rearrangement that provides α-linked glycosylation products with a free C2-hydroxyl. Accessibility to these compounds may further advance glycolipidomic profiling of immune modulating bacterial derived-glycans.
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Affiliation(s)
- Ryan A Davis
- Department of Chemistry, University of California, Davis, One Shields Avenue, Davis, CA 95616, United States
| | - James C Fettinger
- Department of Chemistry, University of California, Davis, One Shields Avenue, Davis, CA 95616, United States
| | - Jacquelyn Gervay-Hague
- Department of Chemistry, University of California, Davis, One Shields Avenue, Davis, CA 95616, United States
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Tandem glycosyl iodide glycosylation and regioselective enzymatic acylation affords 6-O-tetradecanoyl-α-d-cholesterylglycosides. J Org Chem 2014; 79:8447-52. [PMID: 25093454 PMCID: PMC4156253 DOI: 10.1021/jo501371h] [Citation(s) in RCA: 16] [Impact Index Per Article: 1.5] [Reference Citation Analysis] [Abstract] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 01/16/2023]
Abstract
![]()
A generalized synthesis of α-d-cholesterylglycosides
has been achieved using one-pot per-O-trimethylsilyl
glycosyl iodide glycosidation. Both cholesteryl α-d-glucopyranoside (αCG) and cholesteryl α-d-galactopyranoside
were prepared in high yield. These compounds were further esterified
using regioselective enzymatic acylation with tetradecanoyl vinyl
ester to afford 6-O-tetradecanoyl-α-d-cholesteryl glucopyranoside (αCAG) of Helicobacter
pylori and the corresponding galactose analogue in 66–78%
overall yields from free sugars. The tandem step-economy sequence
provides novel analogues to facilitate glycolipidomic profiling.
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Gonçalves IC, Henriques PC, Seabra CL, Martins MCL. The potential utility of chitosan micro/nanoparticles in the treatment of gastric infection. Expert Rev Anti Infect Ther 2014; 12:981-92. [DOI: 10.1586/14787210.2014.930663] [Citation(s) in RCA: 40] [Impact Index Per Article: 3.6] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 12/14/2022]
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Inngjerdingen KT, Thöle C, Diallo D, Paulsen BS, Hensel A. Inhibition of Helicobacter pylori adhesion to human gastric adenocarcinoma epithelial cells by aqueous extracts and pectic polysaccharides from the roots of Cochlospermum tinctorium A. Rich. and Vernonia kotschyana Sch. Bip. ex Walp. Fitoterapia 2014; 95:127-32. [DOI: 10.1016/j.fitote.2014.03.009] [Citation(s) in RCA: 22] [Impact Index Per Article: 2.0] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 08/26/2013] [Revised: 03/05/2014] [Accepted: 03/09/2014] [Indexed: 02/08/2023]
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8
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Magen E, Delgado JS. Helicobacter pylori and skin autoimmune diseases. World J Gastroenterol 2014; 20:1510-1516. [PMID: 24587626 PMCID: PMC3925859 DOI: 10.3748/wjg.v20.i6.1510] [Citation(s) in RCA: 42] [Impact Index Per Article: 3.8] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Download PDF] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Received: 09/21/2013] [Revised: 11/04/2013] [Accepted: 01/06/2014] [Indexed: 02/06/2023] Open
Abstract
Autoimmune skin diseases are characterized by dysregulation of the immune system resulting in a loss of tolerance to skin self-antigen(s). The prolonged interaction between the bacterium and host immune mechanisms makes Helicobacter pylori (H. pylori) a plausible infectious agent for triggering autoimmunity. Epidemiological and experimental data now point to a strong relation of H. pylori infection on the development of many extragastric diseases, including several allergic and autoimmune diseases. H. pylori antigens activate cross-reactive T cells and induce autoantibodies production. Microbial heat shock proteins (HSP) play an important role of in the pathogenesis of autoimmune diseases because of the high level of sequence homology with human HSP. Eradication of H. pylori infection has been shown to be effective in some patients with chronic autoimmune urticaria, psoriasis, alopecia areata and Schoenlein-Henoch purpura. There is conflicting and controversial data regarding the association of H. pylori infection with Behçet’s disease, scleroderma and autoimmune bullous diseases. No data are available evaluating the association of H. pylori infection with other skin autoimmune diseases, such as vitiligo, cutaneous lupus erythematosus and dermatomyositis. The epidemiological and experimental evidence for a possible role of H. pylori infection in skin autoimmune diseases are the subject of this review.
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Ito Y, Vela JL, Matsumura F, Hoshino H, Tyznik A, Lee H, Girardi E, Zajonc DM, Liddington R, Kobayashi M, Bao X, Bugaytsova J, Borén T, Jin R, Zong Y, Seeberger PH, Nakayama J, Kronenberg M, Fukuda M. Helicobacter pylori cholesteryl α-glucosides contribute to its pathogenicity and immune response by natural killer T cells. PLoS One 2013; 8:e78191. [PMID: 24312443 PMCID: PMC3846475 DOI: 10.1371/journal.pone.0078191] [Citation(s) in RCA: 49] [Impact Index Per Article: 4.1] [Reference Citation Analysis] [Abstract] [MESH Headings] [Grants] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 05/28/2013] [Accepted: 09/09/2013] [Indexed: 12/31/2022] Open
Abstract
Approximately 10–15% of individuals infected with Helicobacter pylori will develop ulcer disease (gastric or duodenal ulcer), while most people infected with H. pylori will be asymptomatic. The majority of infected individuals remain asymptomatic partly due to the inhibition of synthesis of cholesteryl α-glucosides in H. pylori cell wall by α1,4-GlcNAc-capped mucin O-glycans, which are expressed in the deeper portion of gastric mucosa. However, it has not been determined how cholesteryl α-glucosyltransferase (αCgT), which forms cholesteryl α-glucosides, functions in the pathogenesis of H. pylori infection. Here, we show that the activity of αCgT from H. pylori clinical isolates is highly correlated with the degree of gastric atrophy. We investigated the role of cholesteryl α-glucosides in various aspects of the immune response. Phagocytosis and activation of dendritic cells were observed at similar degrees in the presence of wild-type H. pylori or variants harboring mutant forms of αCgT showing a range of enzymatic activity. However, cholesteryl α-glucosides were recognized by invariant natural killer T (iNKT) cells, eliciting an immune response in vitro and in vivo. Following inoculation of H. pylori harboring highly active αCgT into iNKT cell-deficient (Jα18−/−) or wild-type mice, bacterial recovery significantly increased in Jα18−/− compared to wild-type mice. Moreover, cytokine production characteristic of Th1 and Th2 cells dramatically decreased in Jα18−/− compared to wild-type mice. These findings demonstrate that cholesteryl α-glucosides play critical roles in H. pylori-mediated gastric inflammation and precancerous atrophic gastritis.
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Affiliation(s)
- Yuki Ito
- Cancer Center, Sanford-Burnham Medical Research Institute, La Jolla, California, United States of America
| | - Jose Luis Vela
- La Jolla Institute for Allergy & Immunology, La Jolla, California, United States of America
| | - Fumiko Matsumura
- Cancer Center, Sanford-Burnham Medical Research Institute, La Jolla, California, United States of America
| | - Hitomi Hoshino
- Department of Molecular Pathology, Shinshu University Graduate School of Medicine, Matsumoto, Nagano, Japan
| | - Aaron Tyznik
- La Jolla Institute for Allergy & Immunology, La Jolla, California, United States of America
| | - Heeseob Lee
- Cancer Center, Sanford-Burnham Medical Research Institute, La Jolla, California, United States of America
| | - Enrico Girardi
- La Jolla Institute for Allergy & Immunology, La Jolla, California, United States of America
| | - Dirk M. Zajonc
- La Jolla Institute for Allergy & Immunology, La Jolla, California, United States of America
| | - Robert Liddington
- Infectious and Inflammatory Disease Center, Sanford-Burnham Medical Research Institute, La Jolla, California, United States of America
| | - Motohiro Kobayashi
- Department of Molecular Pathology, Shinshu University Graduate School of Medicine, Matsumoto, Nagano, Japan
| | - Xingfeng Bao
- Cancer Center, Sanford-Burnham Medical Research Institute, La Jolla, California, United States of America
| | - Jeanna Bugaytsova
- Department of Medical Biochemistry and Biophysics, Umeå University, Umeå, Sweden
| | - Thomas Borén
- Department of Medical Biochemistry and Biophysics, Umeå University, Umeå, Sweden
| | - Rongsheng Jin
- Del E. Webb Neuroscience, Aging and Stem Cell Research Center, Sanford-Burnham Medical Research Institute, La Jolla, California, United States of America
| | - Yinong Zong
- Del E. Webb Neuroscience, Aging and Stem Cell Research Center, Sanford-Burnham Medical Research Institute, La Jolla, California, United States of America
| | - Peter H. Seeberger
- Cancer Center, Sanford-Burnham Medical Research Institute, La Jolla, California, United States of America
| | - Jun Nakayama
- Department of Molecular Pathology, Shinshu University Graduate School of Medicine, Matsumoto, Nagano, Japan
| | - Mitchell Kronenberg
- La Jolla Institute for Allergy & Immunology, La Jolla, California, United States of America
| | - Minoru Fukuda
- Cancer Center, Sanford-Burnham Medical Research Institute, La Jolla, California, United States of America
- * E-mail:
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Gonçalves IC, Magalhães A, Fernandes M, Rodrigues IV, Reis CA, Martins MCL. Bacterial-binding chitosan microspheres for gastric infection treatment and prevention. Acta Biomater 2013; 9:9370-8. [PMID: 23920152 DOI: 10.1016/j.actbio.2013.07.034] [Citation(s) in RCA: 25] [Impact Index Per Article: 2.1] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 03/25/2013] [Revised: 07/17/2013] [Accepted: 07/28/2013] [Indexed: 01/09/2023]
Abstract
Helicobacter pylori (H. pylori) colonizes the gastric mucosa of over 50% of the world population, causing several pathologies, such as gastric ulcers and gastric cancer. Since current antibiotic treatments are inefficient in 20% of cases alternative therapies are needed. This work reports the ability of chitosan microspheres to adhere to H. pylori and prevent/remove H. pylori colonization. Adhesion of H. pylori strains with different functional adhesins (BabA and/or SabA) to chitosan microspheres (diameter 167 ± 27 μm) occurs at both pH 2.6 and 6.0, but is higher at pH 6.0. Bacterial adhesion to a gastric cell line expressing sialylated carbohydrates (SabA receptors) was performed at the same pH values using H. pylori strains with and without SabA. At both pH values addition of microspheres to gastric cells before and after pre-incubation with H. pylori decreased bacterial adhesion to cells. Furthermore, the chitosan microspheres were non-cytotoxic. These findings reveal the potential of chitosan microspheres as an alternative or complementary treatment for H. pylori gastric eradication or prevention of H. pylori colonization.
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Affiliation(s)
- Inês C Gonçalves
- INEB -Instituto de Engenharia Biomédica, Universidade do Porto, Rua do Campo Alegre, 823, 4150-180 Porto, Portugal; IPATIMUP -Institute of Molecular Pathology and Immunology of the University of Porto, Rua Dr. Roberto Frias, 4200-465 Porto, Portugal; Faculdade de Engenharia, Universidade do Porto, Porto, Portugal
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11
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Abstract
Mucin-type O-glycans are the primary constituents of mucins that are expressed on various mucosal sites of the body, especially the bacteria-laden intestinal tract. Mucins are the main components of mucus, which is secreted by goblet cells and forms a protective homeostatic barrier between the resident microbiota and the underlying immune cells in the colon. However, the specific role of mucin-type O-glycans in mucus barrier function has been uncertain. Recent studies utilizing mice deficient in key glycosyltransferases involved in O-glycan biosynthesis on intestinal mucins have underscored the importance of mucin-type O-glycosylation in mucus barrier function. This review will highlight recent advances in our understanding of mucin-type O-glycan function in the mucus barrier and how they promote mutualism with our resident microbiota.
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Affiliation(s)
- Kirk S B Bergstrom
- Cardiovascular Biology Research Program, Oklahoma Medical Research Foundation, Oklahoma City, OK 73104, USA
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12
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Davis RA, Lin CH, Gervay-Hague J. Chemoenzymatic synthesis of cholesteryl-6-O-tetradecanoyl-α-D-glucopyranoside: a product of host cholesterol efflux promoted by Helicobacter pylori. Chem Commun (Camb) 2012; 48:9083-5. [PMID: 22854787 DOI: 10.1039/c2cc33948j] [Citation(s) in RCA: 25] [Impact Index Per Article: 1.9] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 01/26/2023]
Abstract
In a three-step protocol involving regioselective enzymatic acylation, per-O-trimethylsilylation, and a one-pot α-glycosidation-deprotection sequence, cholesteryl-6-O-tetradecanoyl-α-D-glucopyranoside (α-CAG) of Helicobacter pylori is afforded starting from glucose in an overall yield of 45%. The production of CAG can be scaled to make purified quantities available to the biological community for the first time.
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Affiliation(s)
- Ryan A Davis
- Department of Chemistry, University of California Davis, One Shields Avenue, Davis, CA 95616, USA
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13
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Ferreira JA, Silva L, Monteiro MA, Coimbra* MA. Helicobacter pyloricell-surface glycans structural features: role in gastric colonization, pathogenesis, and carbohydrate-based vaccines. CARBOHYDRATE CHEMISTRY 2011:160-193. [DOI: 10.1039/9781849732765-00160] [Citation(s) in RCA: 6] [Impact Index Per Article: 0.4] [Reference Citation Analysis] [Track Full Text] [Subscribe] [Scholar Register] [Indexed: 01/06/2025]
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14
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Bones J, Byrne JC, O'Donoghue N, McManus C, Scaife C, Boissin H, Nastase A, Rudd PM. Glycomic and glycoproteomic analysis of serum from patients with stomach cancer reveals potential markers arising from host defense response mechanisms. J Proteome Res 2011; 10:1246-65. [PMID: 21142185 DOI: 10.1021/pr101036b] [Citation(s) in RCA: 106] [Impact Index Per Article: 7.6] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 12/11/2022]
Abstract
Despite the reduced incidence of gastric cancer in the developed world, a diagnosis of stomach carcinoma still carries a poor prognosis due to the asymptomatic nature of the disease in the early stages, subsequent advanced stage diagnosis, and a low 5 year survival rate. Endoscopy remains the primary standard for diagnosis of stomach carcinoma and the current marker, carbohydrate antigen 19-9 (CA19-9) lacks the levels of sensitivity and specificity required in order to make it clinically useful for diagnostic monitoring. Therefore, there is a current need for additional markers to improve the diagnostic accuracy for the early stages of stomach cancer. Together, glycomic, proteomic, and glycoproteomic analyses of serum have the potential to identify such probable markers. A discovery study is reported here using preoperative serum from 80 stomach cancer patients, 10 patients bearing benign stomach disease, and 20 matched controls. Glycomic analysis of the total and immunoaffinity depleted serum revealed statistically significant increases in the levels of sialyl Lewis X epitopes (SLe(X)) present on triantennary glycans accompanied by increased levels of core fucosylated agalactosyl biantennary glycans present on IgG (referred to as the IgG G0 glycoform) which are associated with increasing disease pathogenesis. Protein expression analysis using 2D-DiGE returned a number of differentially expressed protein candidates in the depleted serum, many of which were shown to carry triantennary SLe(X) during subsequent glycomic investigations. Biological pathway analysis of the experimental data returned complement activation and acute phase response signaling as the most significantly altered pathways in the stomach cancer patient serum. Upon the basis of these findings, it is suggested that increased expression of IgG G0 and complement activation are a host response to the presence of the stomach tumor while the increased expression of SLe(X) and acute phase response proteins is a result of pro-inflammatory cytokine signaling, including IL-6, during carcinogenesis. The approach presented herein provides an insight into the underlying mechanisms of disease and the resulting changes in the glycome and glycoproteome offer promise as potential markers for diagnosis and prognostic monitoring in stomach cancer.
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Affiliation(s)
- Jonathan Bones
- NIBRT Dublin-Oxford Glycobiology Laboratory, The National Institute for Bioprocessing Research and Training, University College Dublin, Belfield, Dublin 4, Ireland
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15
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Fujita M, Tsuchida A, Hirata A, Kobayashi N, Goto K, Osumi K, Hirose Y, Nakayama J, Yamanoi T, Ashida H, Mizuno M. Glycoside hydrolase family 89 alpha-N-acetylglucosaminidase from Clostridium perfringens specifically acts on GlcNAc alpha1,4Gal beta1R at the non-reducing terminus of O-glycans in gastric mucin. J Biol Chem 2010; 286:6479-89. [PMID: 21177247 DOI: 10.1074/jbc.m110.206722] [Citation(s) in RCA: 26] [Impact Index Per Article: 1.7] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 01/30/2023] Open
Abstract
In mammals, α-linked GlcNAc is primarily found in heparan sulfate/heparin and gastric gland mucous cell type mucin. α-N-acetylglucosaminidases (αGNases) belonging to glycoside hydrolase family 89 are widely distributed from bacteria to higher eukaryotes. Human lysosomal αGNase is well known to degrade heparin and heparan sulfate. Here, we reveal the substrate specificity of αGNase (AgnC) from Clostridium perfringens strain 13, a bacterial homolog of human αGNase, by chemically synthesizing a series of disaccharide substrates containing α-linked GlcNAc. AgnC was found to release GlcNAc from GlcNAcα1,4Galβ1pMP and GlcNAcα1pNP substrates (where pMP and pNP represent p-methoxyphenyl and p-nitrophenyl, respectively). AgnC also released GlcNAc from porcine gastric mucin and cell surface mucin. Because AgnC showed no activity against any of the GlcNAcα1,2Galβ1pMP, GlcNAcα1,3Galβ1pMP, GlcNAcα1,6Galβ1pMP, and GlcNAcα1,4GlcAβ1pMP substrates, this enzyme may represent a specific glycosidase required for degrading α-GlcNAc-capped O-glycans of the class III mucin secreted from the stomach and duodenum. Deletion of the C-terminal region containing several carbohydrate-binding module 32 (CBM32) domains significantly reduced the activity for porcine gastric mucin; however, activity against GlcNAcα1,4Galβ1pMP was markedly enhanced. Dot blot and ELISA analyses revealed that the deletion construct containing the C-terminal CBM-C2 to CBM-C6 domains binds strongly to porcine gastric mucin. Consequently, tandem CBM32 domains located near the C terminus of AgnC should function by increasing the affinity for branched or clustered α-GlcNAc-containing glycans. The agnC gene-disrupted strain showed significantly reduced growth on the class III mucin-containing medium compared with the wild type strain, suggesting that AgnC might have an important role in dominant growth in intestines.
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Affiliation(s)
- Masaya Fujita
- Noguchi Institute, 1-8-1 Kaga, Itabashi, Tokyo 173-0003, Japan.
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Hoang V, Williams M, Simpson H. Monosaccharide composition of fundic and duodenal mucins in sheep infected with Haemonchus contortus or Teladorsagia circumcincta. Vet Parasitol 2010; 170:253-61. [DOI: 10.1016/j.vetpar.2010.02.014] [Citation(s) in RCA: 7] [Impact Index Per Article: 0.5] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 11/06/2009] [Revised: 02/01/2010] [Accepted: 02/10/2010] [Indexed: 02/03/2023]
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