New strategy of protein expression in mammalian cells for in-cell NMR

The research activities of my doctorate have been devoted to the application of in-cell NMR for the characterization of proteins directly inside human cells. Within this context, I have also addressed the development of a methodology to perform sequential labeling for in-cell NMR in human cells. The in-cell NMR approach, which consists in observing one or more selectively labeled protein(s) or nucleic acid in living cells through high resolution NMR experiments, is a powerful tool to obtain structural and functional information in situ, thus overcoming the limitation of studying macromolecules isolated from the other cellular components. Protein folding, posttranslational modifications and interactions with specific partners or cellular elements can be studied in the physiological environment in which the protein of interest operates. My research work also addressed the in vitro characterization of the phosphorylation pattern of the non-structural protein 5A domain 3 of Hepatitis C virus by casein kinase 2.