Sato Chihiro, Yamakawa Nao, Kitajima Ken
Bioscience and Biotechnology Center, Graduate School of Bioagricultural Sciences, Nagoya University, Nagoya, Japan.
Methods Enzymol. 2010;478:219-32. doi: 10.1016/S0076-6879(10)78010-1.
Proteins and lipids are often modified with glycan chains, which due to their large hydration effect and structural heterogeneity, significantly alter the surface physicochemical properties of proteins and biomembranes. This "glyco-atmosphere" also serves as a field for interactions with various molecules, including other glycans, lipids, peptides, proteins, and small molecules such as neurotransmitters and drugs as well as lectins. Therefore, sensitive techniques for measuring these glycan-based interactions are becoming more and more necessary, with the appropriate method largely depending on the interacting molecules. In this chapter, we focus on frontal affinity chromatography (FAC) and surface plasmon resonance (SPR) for examining polysialic acid-involved interactions with neurotransmitters and neurotrophins. FAC is characterized by its applicability to analyze weak interactions that are difficult to measure using conventional methods, and by the ease of principle and experimental procedures. SPR is advantageous due to the availability of suitable surface materials and for real-time monitoring with nonlabeled analytes.
蛋白质和脂质常常会被糖链修饰，由于糖链具有较大的水合效应和结构异质性，会显著改变蛋白质和生物膜的表面物理化学性质。这种“糖氛围”还充当了与各种分子相互作用的场所，这些分子包括其他聚糖、脂质、肽、蛋白质以及诸如神经递质、药物等小分子，还有凝集素。因此，用于测量这些基于聚糖的相互作用的灵敏技术变得越来越必要，而合适的方法在很大程度上取决于相互作用的分子。在本章中，我们重点介绍前沿亲和色谱法（FAC）和表面等离子体共振（SPR），用于研究多唾液酸与神经递质和神经营养因子之间的相互作用。FAC的特点在于它适用于分析难以用传统方法测量的弱相互作用，且原理和实验步骤简单。SPR则具有优势，因为有合适的表面材料可用，并且能够对未标记的分析物进行实时监测。
