NMR spectroscopy: from basic concepts to advanced methods

This chapter aims to give a general overview of the field of NMR spectroscopy as applied to structural biology. In the first section the basic concepts of NMR theory are outlined, together with some technical aspects, from the nuclear spin to multidimensional experiments. In the second section more advanced applications of protein NMR are presented, including methods for studying large proteins, fast-pulsing techniques, carbon-detected NMR, paramagnetic NMR and solid-state NMR. The final section focuses on a recent advance of NMR applied to the structural biology of proteins: in-cell NMR. With this method, high-resolution NMR is used to investigate proteins in living cells at atomic detail. The main effects of the intracellular environment on protein structure and dynamics are explained. Several in-cell NMR approaches are shown: protein expression in bacterial cells, protein insertion or expression into higher eukaryotes. Some applications of protein expression by transient transfection in human cells are given.