title: Functional evaluation of carbohydrate-centred glycoclusters by enzyme-linked lectin assay: ligands for concanavalin A.
abstract: The affinities of the mannose-specific lectin concanavalin A (Con A) towards D-glucose-centred mannosyl clusters differing in the anomeric configuration of the monosaccharide core, nature of the bridging functional groups and valency, have been measured by a competitive enzyme-linked lectin assay. Pentavalent thioether-linked ligands (5 and 7) were prepared by radical addition of 2,3,4,6-tetra-O-acetyl-1-thio-alpha-D-mannopyranose to the corresponding penta-O-allyl-alpha- or -beta-D-glucopyranose, followed by deacetylation. The distinct reactivity of the anomeric position in the D-glucose scaffold was exploited in the preparation of a tetravalent cluster (10) that keeps a reactive aglyconic group for further manipulation, including incorporation of a reporter group or attachment to a solid support. Hydroboration of the double bonds in the penta-O-allyl-alpha-D-glucopyranose derivative and replacement of the hydroxy groups with amine moieties gave a suitable precursor for the preparation of pentavalent and 15-valent mannosides through the thiourea-bridging reaction (17 and 20, respectively). The diastereomeric 1-thiomannose-coated clusters 5 and 7 were demonstrated to be potent ligands for Con A, with IC(50) values for the inhibition of the Con A-yeast mannan association indicative of 6.4- and 5.5-fold increases in binding affinity (valency-corrected values), respectively, relative to the value for methyl alpha-D-mannopyranoside. The tetravalent cluster 10 exhibited a valency-corrected relative lectin-binding potency virtually identical to that of the homologous pentavalent mannoside 7. In sharp contrast, replacement of the 1-thiomannose wedges of 5 with alpha-D-mannopyranosylthioureido units (17) virtually abolished any multivalent or statistic effects, with a dramatic decrease of binding affinity. The 15-valent ligand 20, possessing classical O-glycosidic linkages, exhibited a twofold increase in lectin affinity relative to the penta-O-(thioglycoside) 5; it is less efficient based on the number of mannose units. The results illustrate the potential of carbohydrates as polyfunctional platforms for glycocluster construction and underline the importance of careful design of the overall architecture in optimising glycocluster recognition by specific lectins.
authors.affiliation: Instituto de Investigaciones Químicas, CSIC, Américo Vespucio 49, Isla de la Cartuja, 41092 Seville, Spain.
authors.forename: Maja
authors.initials: M
authors.lastname: Köhn
authors.forename: Juan M
authors.initials: JM
authors.lastname: Benito
authors.forename: Carmen
authors.initials: C
authors.lastname: Ortiz Mellet
authors.forename: Thisbe K
authors.initials: TK
authors.lastname: Lindhorst
authors.forename: José M
authors.initials: JM
authors.lastname: García Fernández
mesh_terms: D001665:Binding Sites; D002236:Carbohydrate Conformation; D050260:Carbohydrate Metabolism; D002240:Carbohydrate Sequence; D002241:Carbohydrates / Q000737:chemistry; D003208:Concanavalin A / Q000378:metabolism*; D005947:Glucose / Q000737:chemistry / Q000378:metabolism; D006027:Glycosides / Q000737:chemistry*; D037102:Lectins / Q000737:chemistry / Q000378:metabolism*; D008024:Ligands; D008358:Mannose / Q000737:chemistry / Q000378:metabolism*; D013237:Stereoisomerism; D013890:Thiourea / Q000737:chemistry
chemical_list: D002241:Carbohydrates; D006027:Glycosides; D037102:Lectins; D008024:Ligands; D003208:Concanavalin A; D013890:Thiourea; D005947:Glucose; D008358:Mannose