Characterization of intrinsically disordered proteins by nuclear magnetic resonance spectroscopy

During my doctorate, I focused my studies on the development of NMR methods for the characterization of IDPs and on their application to study challenging cases. The combination of 13C detection with new hardware tools such as Multiple Receivers, and new software tools such as Automated Projection, led to new strategies to speed up NMR experiments. Moreover, modification of base pulse sequences led to simple 13C NMR experiments for the characterization of proline residues, that are highly abundant in IDPs and less straightforward to be investigated with standard HN-based NMR experiments. Together with the development of new methods, I worked on their application to study different aspects of IDPs function.NMR spectroscopy was used for the characterization of a large viral disordered protein, and the study of an unexpected liquid-liquid phase transition likely relevant in the biological mechanism of viral infection. Exploiting NMR we were able to study the interaction between a disordered protein involved in Parkinson’s disease and a small molecule that can be a potential drug to contrast the pathology, as well as the interaction between an oncogenic viral protein and the disordered region of a transcriptional coactivator. These interesting applications demonstrate once again the importance of NMR spectroscopy for studying IDPs, adding small but important pieces in the knowledge about the role of protein disorder in biology.