Most of the structural and functional information about biological macromolecules is obtained from experiments in vitro, far from their physiological context. Elucidating the structure, dynamics, function and molecular interactions of such complex molecules in their cellular context is necessary to understand physiological processes. NMR spectroscopy is an upcoming technique to investigate structural and dynamical features of macromolecules at atomic resolution in living cells. To date, in-cell solution and solid-state NMR approaches have been successfully applied to both prokaryotes and eukaryotic cells to study the structure and dynamical behavior of macromolecules in their natural environment .
We use solid-state NMR in combination with biosynthetic uniformly 13C isotope labeling to elucidate chemical profiles, molecular structure and dynamics of photosynthetic membranes in functional states in intact Chlamydomonas reinhardtii algae cells. Wild-type and mutant cells are used to follow structural-dynamic changes that correlate with physiological membrane responses in the cell. The morphology of cells and the oxygen production activity are checked after each NMR experiment. This unique approach enables us to gain more insight on the characteristics of thylakoid membrane by simultaneously detecting proteins and lipids and following their molecular dynamics directly inside the living cell.
1.Luchinat, E.; Banci, L. IUCrJ 2017, 4, 108.