Kondoh Yasumitsu, Honda Kaori, Osada Hiroyuki
Antibiotics Laboratory, RIKEN, 2-1 Hirosawa, Wako, Saitama, 351-0198, Japan.
Methods Mol Biol. 2015;1263:29-41. doi: 10.1007/978-1-4939-2269-7_3.
Chemical array technology is a powerful tool for high-throughput screening of small-molecule ligand-protein interactions. A chemical array is a collection of small-molecule compounds spotted and immobilized on a glass slide surface, providing a multiplex platform to identify small-molecule compounds binding to a protein of interest in high-throughput screening. Several research groups have developed a variety of methods for the immobilization of small molecules onto a solid matrix. We have developed a unique photo-cross-linked chemical array for immobilizing small molecules in a functional-group-independent manner. In this chapter, we describe in detail a protocol for the construction of a photo-cross-linked chemical array and its application for ligand screening by using a tag-fused protein.
化学阵列技术是用于高通量筛选小分子配体-蛋白质相互作用的强大工具。化学阵列是点样并固定在载玻片表面的小分子化合物的集合,为在高通量筛选中鉴定与目标蛋白质结合的小分子化合物提供了一个多重平台。几个研究小组已经开发出了多种将小分子固定到固体基质上的方法。我们已经开发出一种独特的光交联化学阵列,用于以不依赖官能团的方式固定小分子。在本章中,我们详细描述了构建光交联化学阵列的方案及其通过使用标签融合蛋白进行配体筛选的应用。