HOPPI-NMR: Hot-Peptide Based Screening Assay for Inhibitor of Protein-Protein Interactions by NMR

The contribution of Protein–Protein Interactions (PPIs) in the onset and/or progression of several diseases, especially cancer, have paved the way for considering the PPIs disruption as attractive antitumor strategy. In this context, simple and efficient biophysical methods for detecting the interaction of the inhibitors with the protein counterpart are still in high demand. Herein, we describe a convenient NMR method for the screening of putative PPI inhibitors based on the use of “hot peptides” (HOPPI-NMR). As a case study, HOPPI-NMR was successful applied to the well-known p53/MDM2 system. Our outcomes highlighted the main advantages of the method, including the use of little amount of unlabeled proteins, the minimization of the risk of protein aggregation and the possibility to identify weak binders. The latter opens towards the application of HOPPI-NMR in tandem with fragment-based drug discovery (FBDD) as valid strategy for the identification of novel chemotypes acting as PPIs inhibitors