Resolution of chemical shift anisotropy in 19F ENDOR spectroscopy at 263 GHz/9.4 T

Pulsed F-19 ENDOR spectroscopy provides a selective method for measuring angstrom to nanometer distances in structural biology. Here, the performance of F-19 ENDOR at fields of 3.4 T and 9.4 T is compared using model compounds containing one to three F-19 atoms. CF3 groups are included in two compounds, for which the possible occurrence of uniaxial rotation might affect the distance distribution. At 9.4 T, pronounced asymmetric features are observed in many of the presented F-19 ENDOR spectra. Data analysis by spectral simulations shows that these features arise from the chemical shift anisotropy (CSA) of the F-19 nuclei. This asymmetry is also observed at 3.4 T, albeit to a much smaller extent, confirming the physical origin of the effect. The CSA parameters are well consistent with DFT predicted values and can be extracted from simulation of the experimental data in favourable cases, thereby providing additional information about the geometrical and electronic structure of the spin system. The feasibility of resolving the CSA at 9.4 T provides important information for the interpretation of line broadening in ENDOR spectra also at lower fields, which is relevant for developing methods to extract distance distributions from F-19 ENDOR spectra. (C) 2021 The Authors. Published by Elsevier Inc.

Automated summary

Pulsed F-19 ENDOR spectroscopy is a selective method for measuring distances in structural biology, with pronounced asymmetric features observed at 9.4 T due to chemical shift anisotropy. CSA parameters are consistent with DFT predicted values and can be extracted from experimental data simulations.