The understanding of protein-ligand binding interactions is of fundamental importance in mechanistic enzymology and inhibitor discovery. Nuclear magnetic resonance (NMR) spectroscopy is an established biophysical technique that allows the monitoring and quantification of these dynamic binding events in solution, which can also provide information including kinetics, structural and conformational changes. However, NMR experiments typically suffer from low intrinsic sensitivity and may require careful optimisation of experimental conditions. These factors may limit the applicability of NMR spectroscopy in protein studies.
There exists many NMR experiments, but fundamentally they can be divided into two approaches. One involves observing the signals of a ligand (“ligand-observe NMR”) and the other to observe the signals of the protein (“protein-observe NMR”). I am interested in the structural, mechanistic and inhibition studies of proteins and enzymes, and in the last eight years I have developed and applied both protein-observe and ligand-observe NMR spectroscopy techniques to study protein-ligand binding and enzyme kinetics. In this talk I will review these NMR methods, and demonstrate the feasibility and highlight the approaches that we applied to overcome some of the technical challenges of applying NMR spectroscopy for inhibitor screening with examples from our work with different enzyme systems.