Determination of Protein–Ligand Binding Modes Using Complexation-Induced Changes in 1H NMR Chemical Shift

A new method for determining three-dimensional solution structures of protein–ligand complexes using experimentally determined complexation-induced changes in 1H NMR chemical shift (CIS) is introduced. The method has been validated using the complex formed between the protein antitumor antibiotic neocarzinostatin (NCS) and a synthetic chromophore analogue. The X-ray crystal structure of the unbound protein and the backbone amide proton CIS were the input data used in the determination of the three-dimensional structure of the complex. The experimental CIS values were used in a continuous direct structure refinement process based on genetic algorithms to sample conformational space. The calculated structure of the complex agrees well with the NMR solution structure, indicating the potential of this approach for structure determination.