a Université de Lyon, CNRS, Université Claude Bernard Lyon 1, ENS Lyon, CRMN FRE 2034 , Villeurbanne , France.
b Université de Lyon, Université Claude Bernard Lyon 1, Faculté de Pharmacie - ISPB, EA 4446 Bioactive Molecules and Medicinal Chemistry, SFR Santé Lyon-Est CNRS UMS3453 - INSERM US7 , Lyon , France.
J Enzyme Inhib Med Chem. 2019 Dec;34(1):1218-1225. doi: 10.1080/14756366.2019.1636235.
WaterLOGSY is a sensitive ligand-observed NMR experiment for detection of interaction between a ligand and a protein and is now well-established as a screening technique for fragment-based lead discovery. Here we develop and assess a protocol to derive ligand epitope mapping from WaterLOGSY data and demonstrate its general applicability in studies of fragment-sized ligands binding to six different proteins (glycogen phosphorylase, protein peroxiredoxin 5, Bcl-x, Mcl-1, HSP90, and human serum albumin). We compare the WaterLOGSY results to those obtained from the more widely used saturation transfer difference experiments and to the 3D structures of the complexes when available. In addition, we evaluate the impact of ligand labile protons on the WaterLOGSY data. Our results demonstrate that the WaterLOGSY experiment can be used as an additional confirmation of the binding mode of a ligand to a protein.
WaterLOGSY 是一种灵敏的配体观察 NMR 实验，用于检测配体与蛋白质之间的相互作用，现已成为基于片段的先导化合物发现的筛选技术。在这里，我们开发并评估了一种从 WaterLOGSY 数据中得出配体表位图谱的方案，并证明其在研究与六种不同蛋白质（糖原磷酸化酶、蛋白过氧化物酶 5、Bcl-x、Mcl-1、HSP90 和人血清白蛋白）结合的片段大小配体中的普遍适用性。我们将 WaterLOGSY 结果与更广泛使用的饱和转移差异实验的结果以及复合物的 3D 结构进行比较（当有可用结构时）。此外，我们还评估了配体不稳定质子对 WaterLOGSY 数据的影响。我们的结果表明，WaterLOGSY 实验可用作配体与蛋白质结合模式的附加确认。
