DEEP‐STD NMR is a recently developed powerful approach for structure and pharmacophore elucidation of weak protein‐ligand interactions, reporting key information on the orientation of the ligand and the architecture of the binding pocket. The method relies on selective saturation of protein residues in the binding site and the generation of a differential epitope map by observing the ligand, which depicts the nature of the protein residues contacting the ligand in the bound state. Selective saturation requires knowledge of the chemical shift assignment of the protein residues, which can be obtained either experimentally by NMR or predicted from 3D structures. Here we propose a simple experimental procedure to expand the DEEP‐STD NMR methodology to protein‐ligand cases where the spectral assignment of the protein is not available. This is achieved by experimentally identifying the chemical shifts of the residues present in binding hot‐spots on the surface of the receptor protein using 2D NMR experiments combined with addition of a paramagnetic probe.
|Date made available||2018|
|Publisher||University of East Anglia|
|Date of data production||31 Oct 2018|