Synthesis of tetravalent LacNAc-glycoclusters as high-affinity cross-linker against Erythrina cristagalli agglutinin

Makoto Ogata; Yasushi Chuma; Yoshinori Yasumoto; Takashi Onoda; Myco Umemura; Taichi Usui; Enoch Y Park

doi:10.1016/j.bmc.2015.11.026

Synthesis of tetravalent LacNAc-glycoclusters as high-affinity cross-linker against Erythrina cristagalli agglutinin

Bioorg Med Chem. 2016 Jan 1;24(1):1-11. doi: 10.1016/j.bmc.2015.11.026. Epub 2015 Nov 22.

Authors

Makoto Ogata¹, Yasushi Chuma², Yoshinori Yasumoto², Takashi Onoda³, Myco Umemura⁴, Taichi Usui⁵, Enoch Y Park⁶

Affiliations

¹ 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.
² Department of Applied Biological Chemistry, Faculty of Agriculture, Shizuoka University, 836 Ohya, Suruga-ku, Shizuoka 422-8529, Japan.
³ Department of Chemistry and Biochemistry, National Institute of Technology, Fukushima College, 30 Nagao, Iwaki, Fukushima 970-8034, Japan.
⁴ 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.
⁵ Integrated Bioscience Research Division, Graduate School of Science and Technology, Shizuoka University, 836 Ohya, Suruga-ku, Shizuoka 422-8529, Japan.
⁶ 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.

PMID: 26672510
DOI: 10.1016/j.bmc.2015.11.026

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.

Keywords: Carbohydrates; Cluster effect; Lectins; Multivalency.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Chemical Precipitation
Cross-Linking Reagents / chemical synthesis*
Cross-Linking Reagents / chemistry
Entropy
Erythrina
Hemagglutination
Lactose / analogs & derivatives*
Lactose / chemical synthesis*
Lactose / chemistry
Ligands
Molecular Dynamics Simulation
Molecular Structure
Particle Size
Plant Lectins / antagonists & inhibitors*
Plant Lectins / chemistry

Substances

Cross-Linking Reagents
Ligands
Plant Lectins
erythrina lectin
Lactose