Book Chapter

Ligand‐Observed NMR in Fragment‐Based Approaches

Abell, Chris, Śledź, Paweł, Ciulli, Alessio

2012

Overview

One‐dimensional 1 H NMR experiments are commonly employed in both industry and academia for fragment‐based screening against biomolecular protein targets. The overall objective of this chapter is to familiarize the reader with the principles of ligand‐based NMR binding experiments, including an understanding of the theory behind each experiment, and to describe in detail the application of these methods to fragment screening and hit validation. First, some of the theoretical principles of ligand‐observed NMR techniques and their advantages relative to protein‐observed NMR methods are briefly illustrated. We then describe how NMR experiments are used to obtain: (i) yes/no binding answers, (ii) measurements of relative affinities, and (iii) information on binding modes. Finally, practical considerations on performing actual experiments, and processing and analyzing the data are highlighted. The chapter focuses on NMR sequences employing relaxation‐edited filters, saturation transfer difference (STD), water‐ligand observed by gradient spectroscopy (WaterLOGSY), and the interligand Overhauser effect (ILOE), which are mainly employed in our laboratories. Other related ligand‐observed NMR approaches are also described briefly. We especially emphasize our experiences and lessons learnt over the past few years on implementing and applying these techniques to fragment screening and fragment‐based discovery efforts in an academic setup.

• Publisher: Wiley‐VCH Verlag GmbH & Co. KGaA
• Subject: binding mode / fragment‐based design / interligand Overhauser effect / ligand‐observed NMR / relative affinity / relaxation‐edited filter / saturation transfer difference / WaterLOGSY
• ISBN: 3527328505, 9783527328505