Two-dimensional 19F–13C correlation NMR for 19F resonance assignment of fluorinated proteins
Name
nihms-1605328.pdf
Description
Accepted version
Size
2.49 MB
Format
Adobe PDF
Checksum (MD5)
8a52eae833fa3923ea8a6dbbdbbb3fe4
Author(s)
Shcherbakov, Alexander A
Roos, Matthias
Kwon, Byungsu
Hong, Mei
Date Issued
2020
Journal
Journal of Biomolecular NMR
Publisher
Springer Science and Business Media LLC
Version
Author's final manuscript
Abstract
© 2020, Springer Nature B.V. 19F solid-state NMR is an excellent approach for measuring long-range distances for structure determination and for studying molecular motion. For multi-fluorinated proteins, assignment of 19F chemical shifts has been traditionally carried out using mutagenesis. Here we show 2D 19F–13C correlation experiments that allow efficient assignment of the 19F chemical shifts. We have compared several rotational-echo double-resonance-based pulse sequences and 19F–13C cross polarization (CP) for 2D 19F–13C correlation. We found that direct transferred-echo double-resonance (TEDOR) transfer from 19F to 13C and vice versa outperforms out-and-back coherence transfer schemes. 19F detection gives twofold higher sensitivity over 13C detection for the 2D correlation experiment. At MAS frequencies of 25–35 kHz, double-quantum 19F–13C CP has higher coherence transfer efficiencies than zero-quantum CP. The most efficient TEDOR transfer experiment has higher sensitivity than the most efficient double-quantum CP experiment. We demonstrate these 2D 19F–13C correlation experiments on the model compounds t-Boc-4F-phenylalanine and GB1. Application of the 2D 19F–13C TEDOR correlation experiment to the tetrameric influenza BM2 transmembrane peptide shows intermolecular 13C–19F cross peaks that indicate that the BM2 tetramers cluster in the lipid bilayer in an antiparallel fashion. This clustering may be relevant for the virus budding function of this protein.
MIT Department
Massachusetts Institute of Technology. Department of Chemistry
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DOI of Published Version
https://doi.org/10.1007/S10858-020-00306-0
