Ruprecht Colin, Pfrengle Fabian
Department of Biomolecular Systems, Max Planck Institute of Colloids and Interfaces, Potsdam, Germany.
Department of Chemistry, University of Natural Resources and Life Sciences Vienna, Vienna, Austria.
Methods Mol Biol. 2022;2460:115-125. doi: 10.1007/978-1-0716-2148-6_7.
Chemically synthesized plant oligosaccharides have recently evolved as powerful molecular tools for plant cell wall biology. Synthetic plant glycan microarrays equipped with these oligosaccharides enable high-throughput analyses of glycan-binding proteins and carbohydrate-active enzymes. To produce these glycan microarrays, small amounts of glycan solution are printed on suitable surfaces for covalent or non-covalent immobilization. Synthetic plant glycan microarrays have been used for example to map the epitopes of plant cell wall-directed antibodies, to characterize glycosyl hydrolases and glycosyl transferases, and to analyze lectin binding. In this chapter, detailed experimental procedures for the production of synthetic glycan microarrays and their use for the characterization of cell wall glycan-directed antibodies are described.
化学合成的植物寡糖最近已发展成为用于植物细胞壁生物学研究的强大分子工具。配备这些寡糖的合成植物聚糖微阵列能够对聚糖结合蛋白和碳水化合物活性酶进行高通量分析。为了制备这些聚糖微阵列，将少量聚糖溶液印刷在合适的表面上以进行共价或非共价固定。合成植物聚糖微阵列已被用于例如绘制针对植物细胞壁的抗体的表位、表征糖基水解酶和糖基转移酶以及分析凝集素结合。在本章中，描述了合成聚糖微阵列的详细实验步骤及其用于表征细胞壁聚糖导向抗体的方法。
