title: Synthesis of 1-N-glycyl beta-oligosaccharide derivatives. Reactivity of Lens culinaris lectin with a fluorescent labeled streptavidin pseudoglycoprotein and immobilized neoglycolipid.
abstract: The lectin from Lens culinaris (lentil) has a binding specificity for glycopeptides bearing 6-O-linked fucose on the reducing terminus on complex-type N-linked oligosaccharides. Lentil lectin therefore provides an excellent example of a carbohydrate binding protein in which high-affinity interactions are dependent on the integrity of the oligosaccharide core structure. We report here the synthesis of the 1-N-glycyl beta-derivative of Gal beta 4GlcNAc beta 2Man alpha 6(Gal beta 4GlcNAc beta 2Man alpha 3)Man beta 4GlcNAc beta 4(Fuc alpha 6)-GlcNAc (Gal-2F) and its subsequent biotinylation and palmitoylation. The biotin derivative when bound to a streptavidin-fluorescein isothiocyanate (FITC) conjugate was able to bind to both concanavalin A (ConA) and lentil lectin affinity columns. In contrast, synthesis of the biotin derivative of the glycamine derivative of Gal-2F and subsequent binding to streptavidin-FITC afforded reactivity to a ConA affinity column but not to a lentil lectin affinity column. Lentil lectin also bound to plastic microtiter plates containing the adsorbed palmitoyl-1-N-glycyl beta-derivative. No binding occurred when the homologous glycamine neoglycolipid was used. These results suggest the 1-N-glycyl beta-derivative of oligosaccharides may have general utility as an intermediate in the synthesis of novel glycoconjugate probes.
authors.affiliation: Oxford Glycobiology Institute, Department of Biochemistry, University of Oxford, U.K.
authors.forename: I D
authors.initials: ID
authors.lastname: Manger
authors.forename: S Y
authors.initials: SY
authors.lastname: Wong
authors.forename: T W
authors.initials: TW
authors.lastname: Rademacher
authors.forename: R A
authors.initials: RA
authors.lastname: Dwek
mesh_terms: D001426:Bacterial Proteins; D001710:Biotin / Q000737:chemistry; D002236:Carbohydrate Conformation; D002240:Carbohydrate Sequence; D003208:Concanavalin A / Q000378:metabolism; D016650:Fluorescein-5-isothiocyanate; D005456:Fluorescent Dyes; D006017:Glycolipids / Q000378:metabolism*; D006023:Glycoproteins / Q000378:metabolism*; D037102:Lectins / Q000378:metabolism*; D009682:Magnetic Resonance Spectroscopy; D008969:Molecular Sequence Data; D009844:Oligosaccharides / Q000737:chemistry / Q000378:metabolism*; D019308:Palmitic Acid; D010169:Palmitic Acids / Q000737:chemistry; D037121:Plant Lectins; D019809:Streptavidin
chemical_list: D001426:Bacterial Proteins; D005456:Fluorescent Dyes; D006017:Glycolipids; D006023:Glycoproteins; D037102:Lectins; D009844:Oligosaccharides; D010169:Palmitic Acids; D037121:Plant Lectins; C033619:lentil lectin; D003208:Concanavalin A; D019308:Palmitic Acid; D001710:Biotin; D019809:Streptavidin; D016650:Fluorescein-5-isothiocyanate