Aromatic (19)F-(13)C TROSY: A background-free approach to probe biomolecular structure, function and dynamics

Obtaining atomic level information about the structure and dynamics of biomolecules is critical to understand their function. Nuclear magnetic resonance (NMR) spectroscopy provides unique insights into the dynamic nature of biomolecules and their interactions, capturing transient conformers and their features. However, relaxation-induced line broadening and signal overlap make it challenging to apply NMR to large biological systems. Here, we take advantage of the high sensitivity and the broad chemical-shift range of (19)F nuclei, and leverage the remarkable relaxation properties of the aromatic (19)F-(13)C spin pair to disperse (19)F resonances in a 2-dimensional transverse relaxation optimized TROSY spectrum. We demonstrate the application of the (19)F-(13)C TROSY to investigate proteins and nucleic acids. This experiment expands the scope of (19)F NMR in the study of structure, dynamics and function of large and complex biological systems and provides a powerful background-free NMR probe.