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

摘要

Internuclear distances measured using NMR provide crucial constraints of three-dimensional structures, but are often restricted to about 5 Å 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 ¹⁹F spin exchange NMR for precise and robust measurements of inter-atomic distances to 1.6 nm at a magnetic field of 14.1 Tesla, 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 Å for short distances and ±2 Å for long distances. For ¹⁹F spins with the same isotropic chemical shift but different anisotropic chemical shifts, intermediate MAS frequencies of 15 – 25 kHz without ¹H irradiation accelerate spin exchange. For spectrally resolved ¹⁹F-¹⁹F spin exchange, ¹H–¹⁹F dipolar recoupling significantly speeds up ¹⁹F-¹⁹F spin exchange. Based on 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 ¹⁹F-¹⁹F distances to be extracted from the measured spin exchange rates after taking into account ¹⁹F chemical shifts. These results demonstrate the robustness of ¹⁹F spin exchange NMR for distance measurements in a wide range of biological and chemical systems.