Yu Kai, Creagh A Louise, Haynes Charles A, Kizhakkedathu Jayachandran N
Department of Pathology and Laboratory Medicine, Centre for Blood Research, University of British Columbia, Vancouver, BC, Canada, V6T 1Z3.
Michael Smith Laboratories, Department of Chemical and Biological Engineering, University of British Columbia, Vancouver, BC, Canada, V6T 1Z4.
Methods Mol Biol. 2016;1367:183-93. doi: 10.1007/978-1-4939-3130-9_15.
The presentation of carbohydrates on an array can provide a means to model (mimic) oligosaccharides found on cell surfaces. Tuning the structural features of such carbohydrate arrays can therefore be used to help to elucidate the molecular mechanisms of protein-carbohydrate recognition on cell surfaces. Here we present a strategy to directly correlate the molecular and structural features of ligands presented on a surface with the kinetics and affinity of carbohydrate-lectin binding. The Surface Plasmon Resonance (SPR) spectroscopy analysis identified that by varying the spatial distribution (3D organization) of carbohydrate ligands within the surface grafted polymer layer, the mode of binding changed from multivalent to monovalent: a near 1000-fold change in the equilibrium association constant was achieved. The rupture forces measured by atomic force microscopy (AFM) force spectroscopy also indicated that the mode of binding between lectin and carbohydrate ligands can be modulated by the organization of carbohydrate ligands within the glycopolymer brushes.
在阵列上呈现碳水化合物可为模拟（模仿）细胞表面发现的寡糖提供一种手段。因此，调整此类碳水化合物阵列的结构特征可用于帮助阐明细胞表面蛋白质 - 碳水化合物识别的分子机制。在此，我们提出一种策略，将表面呈现的配体的分子和结构特征与碳水化合物 - 凝集素结合的动力学和亲和力直接关联起来。表面等离子体共振（SPR）光谱分析表明，通过改变表面接枝聚合物层内碳水化合物配体的空间分布（三维组织），结合模式从多价变为单价：平衡缔合常数实现了近1000倍的变化。通过原子力显微镜（AFM）力谱测量的断裂力也表明，凝集素与碳水化合物配体之间的结合模式可通过糖聚合物刷内碳水化合物配体的组织来调节。
