Huang Renjie, Leung Ivanhoe K H
School of Chemical Sciences, The University of Auckland, Auckland, New Zealand.
School of Chemical Sciences, The University of Auckland, Auckland, New Zealand.
Methods Enzymol. 2019;615:477-500. doi: 10.1016/bs.mie.2018.08.020. Epub 2018 Sep 20.
WaterLOGSY is a ligand-observed NMR method that is widely used for the studies of protein-small molecule interactions. The basis of waterLOGSY relies on the transfer of magnetization between water molecules, proteins, and small molecules via the nuclear Overhauser effect and chemical exchange. WaterLOGSY is used extensively for the screening of protein ligands, as it is a robust, relatively high-throughput, and reliable method to identify small molecules that bind proteins with a binding affinity (K) in the μM to mM region. WaterLOGSY also enables the determination of K via ligand titration, although careful optimization of the experimental setup is required to avoid overestimation of binding constants. Finally, waterLOGSY allows the water-accessible ligand protons of protein-bound ligands to be identified, thus providing structural information of the ligand binding orientation. In this chapter, we introduce and describe the waterLOGSY method, and provide a practical guide for ligand screening and K determination. The use of waterLOGSY to study water accessibility is also discussed.
水LOGSY是一种配体观测核磁共振方法，广泛应用于蛋白质 - 小分子相互作用的研究。水LOGSY的原理基于水分子、蛋白质和小分子之间通过核Overhauser效应和化学交换实现的磁化转移。水LOGSY被广泛用于蛋白质配体的筛选，因为它是一种稳健、相对高通量且可靠的方法，可用于鉴定与蛋白质结合亲和力（K）在μM至mM范围内的小分子。水LOGSY还可以通过配体滴定法测定K，不过需要仔细优化实验设置以避免高估结合常数。最后，水LOGSY能够识别与蛋白质结合的配体中可与水接触的配体质子，从而提供配体结合取向的结构信息。在本章中，我们介绍并描述水LOGSY方法，并提供配体筛选和K测定的实用指南。还将讨论水LOGSY在研究水可及性方面的应用。
