title: Fucosylated chondroitin sulfates from the body wall of the sea cucumber Holothuria forskali: conformation, selectin binding, and biological activity.
abstract: Fucosylated chondroitin sulfate (fCS) extracted from the sea cucumber Holothuria forskali is composed of the following repeating trisaccharide unit: → 3)GalNAcβ4,6S(1 → 4) [FucαX(1 → 3)]GlcAβ(1 →, where X stands for different sulfation patterns of fucose (X = 3,4S (46%), 2,4S (39%), and 4S (15%)). As revealed by NMR and molecular dynamics simulations, the fCS repeating unit adopts a conformation similar to that of the Le(x) blood group determinant, bringing several sulfate groups into close proximity and creating large negative patches distributed along the helical skeleton of the CS backbone. This may explain the high affinity of fCS oligosaccharides for L- and P-selectins as determined by microarray binding of fCS oligosaccharides prepared by Cu(2+)-catalyzed Fenton-type and photochemical depolymerization. No binding to E-selectin was observed. fCS poly- and oligosaccharides display low cytotoxicity in vitro, inhibit human neutrophil elastase activity, and inhibit the migration of neutrophils through an endothelial cell layer in vitro. Although the polysaccharide showed some anti-coagulant activity, small oligosaccharide fCS fragments had much reduced anticoagulant properties, with activity mainly via heparin cofactor II. The fCS polysaccharides showed prekallikrein activation comparable with dextran sulfate, whereas the fCS oligosaccharides caused almost no effect. The H. forskali fCS oligosaccharides were also tested in a mouse peritoneal inflammation model, where they caused a reduction in neutrophil infiltration. Overall, the data presented support the action of fCS as an inhibitor of selectin interactions, which play vital roles in inflammation and metastasis progression. Future studies of fCS-selectin interaction using fCS fragments or their mimetics may open new avenues for therapeutic intervention.
authors.affiliation: From the EaStCHEM School of Chemistry, Joseph Black Building, The King's Buildings, University of Edinburgh, Edinburgh EH9 3JJ, United Kingdom.
authors.forename: Charalampos G
authors.initials: CG
authors.lastname: Panagos
authors.affiliation: GlycoMar Ltd., European Centre for Marine Biotechnology, Dunstaffnage Marine Laboratory, Oban, Argyll PA37 1QA, United Kingdom.
authors.forename: Derek S
authors.initials: DS
authors.lastname: Thomson
authors.affiliation: GlycoMar Ltd., European Centre for Marine Biotechnology, Dunstaffnage Marine Laboratory, Oban, Argyll PA37 1QA, United Kingdom.
authors.forename: Claire
authors.initials: C
authors.lastname: Moss
authors.affiliation: the Scottish Association for Marine Science, Scottish Marine Institute, Oban, Argyll PA37 1QA, United Kingdom.
authors.forename: Adam D
authors.initials: AD
authors.lastname: Hughes
authors.affiliation: the Scottish Association for Marine Science, Scottish Marine Institute, Oban, Argyll PA37 1QA, United Kingdom.
authors.forename: Maeve S
authors.initials: MS
authors.lastname: Kelly
authors.affiliation: the Glycosciences Laboratory, Hammersmith Campus, Imperial College London, London W12 0NN, United Kingdom.
authors.forename: Yan
authors.initials: Y
authors.lastname: Liu
authors.affiliation: the Glycosciences Laboratory, Hammersmith Campus, Imperial College London, London W12 0NN, United Kingdom.
authors.forename: Wengang
authors.initials: W
authors.lastname: Chai
authors.affiliation: the Sackler Institute of Pulmonary Pharmacology, Institute of Pharmaceutical Science, King's College London, London SE1 9NH, United Kingdom.
authors.forename: Radhakrishnan
authors.initials: R
authors.lastname: Venkatasamy
authors.affiliation: the Sackler Institute of Pulmonary Pharmacology, Institute of Pharmaceutical Science, King's College London, London SE1 9NH, United Kingdom.
authors.forename: Domenico
authors.initials: D
authors.lastname: Spina
authors.affiliation: the Sackler Institute of Pulmonary Pharmacology, Institute of Pharmaceutical Science, King's College London, London SE1 9NH, United Kingdom.
authors.forename: Clive P
authors.initials: CP
authors.lastname: Page
authors.affiliation: the National Institute of Biological Standards and Controls, South Mimms, Potters Bar, Hertfordshire EN6 3QG, United Kingdom.
authors.forename: John
authors.initials: J
authors.lastname: Hogwood
authors.affiliation: the Complex Carbohydrate Research Center, the University of Georgia, Athens, Georgia 30602, and the School of Chemistry, National University of Ireland Galway, University Road, Galway, Ireland.
authors.forename: Robert J
authors.initials: RJ
authors.lastname: Woods
authors.affiliation: the Glycosciences Laboratory, Hammersmith Campus, Imperial College London, London W12 0NN, United Kingdom, the Sackler Institute of Pulmonary Pharmacology, Institute of Pharmaceutical Science, King's College London, London SE1 9NH, United Kingdom.
authors.forename: Barbara
authors.initials: B
authors.lastname: Mulloy
authors.affiliation: GlycoMar Ltd., European Centre for Marine Biotechnology, Dunstaffnage Marine Laboratory, Oban, Argyll PA37 1QA, United Kingdom.
authors.forename: Charlie D
authors.initials: CD
authors.lastname: Bavington
authors.affiliation: From the EaStCHEM School of Chemistry, Joseph Black Building, The King's Buildings, University of Edinburgh, Edinburgh EH9 3JJ, United Kingdom, dusan.uhrin@ed.ac.uk.
authors.forename: Dušan
authors.initials: D
authors.lastname: Uhrín
mesh_terms: D000818:Animals; D000894:Anti-Inflammatory Agents, Non-Steroidal / Q000737:chemistry* / Q000302:isolation & purification / Q000378:metabolism / Q000494:pharmacology; D002236:Carbohydrate Conformation; D002809:Chondroitin Sulfates / Q000737:chemistry* / Q000302:isolation & purification / Q000378:metabolism / Q000494:pharmacology; D006861:Hydrogen Peroxide; D007154:Immune System Diseases / Q000188:drug therapy* / Q000378:metabolism / Q000473:pathology; D007501:Iron; D019041:L-Selectin / Q000737:chemistry / Q000378:metabolism; D007960:Leukocyte Disorders / Q000188:drug therapy* / Q000378:metabolism / Q000473:pathology; D019272:Leukocyte Elastase / Q000037:antagonists & inhibitors / Q000378:metabolism; D051379:Mice; D008958:Models, Molecular; D008969:Molecular Sequence Data; D020556:Neutrophil Infiltration / Q000187:drug effects; D009504:Neutrophils / Q000187:drug effects / Q000378:metabolism / Q000473:pathology; D010084:Oxidation-Reduction; D019007:P-Selectin / Q000737:chemistry / Q000378:metabolism; D010538:Peritonitis / Q000188:drug therapy* / Q000378:metabolism / Q000473:pathology; D053491:Proteinase Inhibitory Proteins, Secretory / Q000737:chemistry* / Q000302:isolation & purification / Q000378:metabolism / Q000494:pharmacology; D012616:Sea Cucumbers / Q000737:chemistry*
chemical_list: D000894:Anti-Inflammatory Agents, Non-Steroidal; C045076:Fenton's reagent; D019007:P-Selectin; D053491:Proteinase Inhibitory Proteins, Secretory; C517150:fucosylated chondroitin sulfate; D019041:L-Selectin; D002809:Chondroitin Sulfates; D006861:Hydrogen Peroxide; D007501:Iron; D019272:Leukocyte Elastase
keywords: Carbohydrate Structure; Carbohydrate-binding Protein; GAG; Glycosaminoglycan; Inflammation; Microarray; Molecular Dynamics; Nuclear Magnetic Resonance (NMR); Selectin