NMR methods for the determination of protein-ligand dissociation constants

The use of NMR to determine quantitatively the dissociation constants of protein-ligand complexes is firmly established. The opportunity to observe cleanly the species of interest with high sensitivity and often with no additional chemical manipulation has resulted in the accumulation of huge experience with NMR. The classical NMR approach to the determination of stoichiometry is the method of continuous variations. An advantage of using NMR to measure protein-ligand interaction is that the NMR method extends the range of measurable interactions into the mM range, a region not well covered by traditional biochemical binding assays. Competition binding and CP-MAS approaches, have resulted in novel, sensitive and specific NMR methods to measure KD that have moved far from the original linewidth and chemical shift perturbation approaches.