title: Synthesis of tetravalent LacNAc-glycoclusters as high-affinity cross-linker against Erythrina cristagalli agglutinin.
abstract: Four kinds of tetravalent double-headed glycoclusters [(LacNAc)4-DHGs] were designed with linkers of varying lengths consisting of alkanedioic carboxyamido groups (C6, C12, C18 and C24) between two bi-antennary LacNAc-glycosides. These glycoclusters served as high-affinity cross-linking ligands for the LacNAc-binding lectin Erythrina cristagalli agglutinin (ECA). The binding activity and cross-linking between each ligand and ECA were characterized by a hemagglutination inhibition (HI) assay, isothermal titration calorimetry (ITC), a quantitative precipitation assay and dynamic light scattering (DLS). For the precipitation assay and DLS measurement, the synthesized (LacNAc)4-DHGs were found to be capable of binding and precipitating the ECA as multivalent ligands. ITC analysis indicated the binding of (LacNAc)4-DHGs was driven by a favorable enthalpy change. Furthermore, the entropy penalty from binding (LacNAc)4-DHGs clearly decreased in a spacer length-dependent manner. The binding affinities of flexible (LacNAc)4-DHGs (C18 and C24) with long spacers were found to be more favorable than those of the clusters having short spacers (C6 and C12). These results were supported by molecular dynamics simulations with explicit water molecules for the tetravalent glycoclusters with ECA. We concluded that the subtle modification in the epitope-presenting scaffolds exerts the significant effect in the recognition efficiency involved in the LacNAc moieties by ECA.
authors.affiliation: Department of Chemistry and Biochemistry, National Institute of Technology, Fukushima College, 30 Nagao, Iwaki, Fukushima 970-8034, Japan. Electronic address: ogata@fukushima-nct.ac.jp.
authors.forename: Makoto
authors.initials: M
authors.lastname: Ogata
authors.affiliation: Department of Applied Biological Chemistry, Faculty of Agriculture, Shizuoka University, 836 Ohya, Suruga-ku, Shizuoka 422-8529, Japan.
authors.forename: Yasushi
authors.initials: Y
authors.lastname: Chuma
authors.affiliation: Department of Applied Biological Chemistry, Faculty of Agriculture, Shizuoka University, 836 Ohya, Suruga-ku, Shizuoka 422-8529, Japan.
authors.forename: Yoshinori
authors.initials: Y
authors.lastname: Yasumoto
authors.affiliation: Department of Chemistry and Biochemistry, National Institute of Technology, Fukushima College, 30 Nagao, Iwaki, Fukushima 970-8034, Japan.
authors.forename: Takashi
authors.initials: T
authors.lastname: Onoda
authors.affiliation: Bioproduction Research Institute, National Institute of Advanced Industrial Science and Technology (AIST), 17-2-1 Higashi-Nijo, Tsukisamu, Toyohira-ku, Sapporo, Hokkaido 062-8517, Japan.
authors.forename: Myco
authors.initials: M
authors.lastname: Umemura
authors.affiliation: Integrated Bioscience Research Division, Graduate School of Science and Technology, Shizuoka University, 836 Ohya, Suruga-ku, Shizuoka 422-8529, Japan.
authors.forename: Taichi
authors.initials: T
authors.lastname: Usui
authors.affiliation: Department of Applied Biological Chemistry, Faculty of Agriculture, Shizuoka University, 836 Ohya, Suruga-ku, Shizuoka 422-8529, Japan; Integrated Bioscience Research Division, Graduate School of Science and Technology, Shizuoka University, 836 Ohya, Suruga-ku, Shizuoka 422-8529, Japan; Research Institute of Green Science and Technology, Shizuoka University, 836 Ohya, Suruga-ku, Shizuoka 422-8529, Japan. Electronic address: park.enoch@shizuoka.ac.jp.
authors.forename: Enoch Y
authors.initials: EY
authors.lastname: Park
mesh_terms: D011232:Chemical Precipitation; D003432:Cross-Linking Reagents / Q000138:chemical synthesis* / Q000737:chemistry; D019277:Entropy; D004895:Erythrina; D006384:Hemagglutination; D007785:Lactose / Q000031:analogs & derivatives* / Q000138:chemical synthesis* / Q000737:chemistry; D008024:Ligands; D056004:Molecular Dynamics Simulation; D015394:Molecular Structure; D010316:Particle Size; D037121:Plant Lectins / Q000037:antagonists & inhibitors* / Q000737:chemistry
chemical_list: D003432:Cross-Linking Reagents; D008024:Ligands; D037121:Plant Lectins; C045629:erythrina lectin; D007785:Lactose
keywords: Carbohydrates; Cluster effect; Lectins; Multivalency