Identification of the Binding Site of an Allosteric Ligand Using STD-NMR, Docking, and CORCEMA-ST Calculations

摘要

Allostery is the coupling of conformational changes between two widely separated sites. It is one of the most common and powerful means to regulate protein function and has been referred to as "the second secret of life". Modulation of the allosteric sites of protein targets provides opportunities for identifying unique molecules with therapeutic advantages over classic active-site inhibitors, such as improved subtype selectivity, decreased drug resistance, and the ability to selectively tune (activate or inhibit) the response of the target protein. With increasing emphasis on cellular functional screens, more allosteric ligands are being discovered as potential drugs. However, discovery and characterization of allosteric sites is still very challenging due to the intrinsic complexity of protein allostery and the experimental difficulties in detecting and verifying allosteric effects and binding sites. Here, we present an easily applicable saturation transfer difference (STD)-NMR/ docking/CORCEMA-ST method for efficient identification and validation of novel allosteric sites.