Fast Magic-Angle-Spinning 19F Spin Exchange NMR for Determining Nanometer 19F-19F Distances in Proteins and Pharmaceutical Compounds

Internuclear distances measured using NMR provide crucial constraints of three-dimensional structures but are often restricted to about 5 angstrom due to the weakness of nuclear-spin dipolar couplings. For studying macromolecular assemblies in biology and materials science, distance constraints beyond 1 nm will be extremely valuable. Here we present an extensive and quantitative analysis of the feasibility of F-19 spin exchange NMR for precise and robust measurements of interatomic distances up to 1.6 nm at a magnetic field of 14.1 T, under 20-40 kHz magic-angle spinning (MAS). The measured distances are comparable to those achievable from paramagnetic relaxation enhancement but have higher precision, which is better than +/- 1 angstrom for short distances and +/- 2 angstrom for long distances. For F-19 spins with the same isotropic chemical shift but different anisotropic chemical shifts, intermediate MAS frequencies of 15-25 kHz without H-1 irradiation accelerate spin exchange. For spectrally resolved F-19-F-19 spin exchange, H-1-F-19 dipolar recoupling significantly speeds up F-19-F-19 spin exchange. On the basis of data from five fluorinated synthetic, pharmaceutical, and biological compounds, we obtained two general curves for spin exchange between CF groups and between CF3, and CF groups. These curves allow F-19-F-19 distances to be extracted from the measured spin exchange rates after taking into account F-19 chemical shifts. These results demonstrate the robustness of F-19 spin exchange NMR for distance measurements in a wide range of biological and chemical systems.