Abstract Limited chemical shift dispersion represents a significant barrier to studying multi-state equilibria of large membrane proteins by 19 F NMR. We describe a novel monofluoroethyl 19 F probe that dramatically increases the chemical shift dispersion. The improved conformational sensitivity and line shape enable the detection of previously unresolved states in 1D NMR spectra of a 134 kDa membrane transporter. Changes in the populations of these states in response to ligand binding, mutations, and temperature correlate with population changes of distinct conformations in structural ensembles determined by single-particle cryo-electron microscopy. Thus, 19 F NMR can guide sample preparation to discover and visualize novel conformational states and facilitate image analysis and 3D classification.
This paper's license is marked as closed access or non-commercial and cannot be viewed on ResearchHub. Visit the paper's external site.