Two-dimensional 19F–13C correlation NMR for 19F resonance assignment of fluorinated proteins
© 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...
| Main Authors: | , , , |
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| Format: | Article |
| Language: | English |
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Springer Science and Business Media LLC
2021
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| Online Access: | https://hdl.handle.net/1721.1/136303 |
| _version_ | 1826190054837452800 |
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| author | Shcherbakov, Alexander A Roos, Matthias Kwon, Byungsu Hong, Mei |
| author2 | Massachusetts Institute of Technology. Department of Chemistry |
| author_facet | Massachusetts Institute of Technology. Department of Chemistry Shcherbakov, Alexander A Roos, Matthias Kwon, Byungsu Hong, Mei |
| author_sort | Shcherbakov, Alexander A |
| collection | MIT |
| description | © 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. |
| first_indexed | 2024-09-23T08:34:21Z |
| format | Article |
| id | mit-1721.1/136303 |
| institution | Massachusetts Institute of Technology |
| language | English |
| last_indexed | 2024-09-23T08:34:21Z |
| publishDate | 2021 |
| publisher | Springer Science and Business Media LLC |
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| spelling | mit-1721.1/1363032023-03-01T20:04:47Z Two-dimensional 19F–13C correlation NMR for 19F resonance assignment of fluorinated proteins Shcherbakov, Alexander A Roos, Matthias Kwon, Byungsu Hong, Mei Massachusetts Institute of Technology. Department of Chemistry © 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. 2021-10-27T20:34:47Z 2021-10-27T20:34:47Z 2020 2020-09-18T14:56:32Z Article http://purl.org/eprint/type/JournalArticle https://hdl.handle.net/1721.1/136303 en 10.1007/S10858-020-00306-0 Journal of Biomolecular NMR Creative Commons Attribution-Noncommercial-Share Alike http://creativecommons.org/licenses/by-nc-sa/4.0/ application/pdf Springer Science and Business Media LLC PMC |
| spellingShingle | Shcherbakov, Alexander A Roos, Matthias Kwon, Byungsu Hong, Mei Two-dimensional 19F–13C correlation NMR for 19F resonance assignment of fluorinated proteins |
| title | Two-dimensional 19F–13C correlation NMR for 19F resonance assignment of fluorinated proteins |
| title_full | Two-dimensional 19F–13C correlation NMR for 19F resonance assignment of fluorinated proteins |
| title_fullStr | Two-dimensional 19F–13C correlation NMR for 19F resonance assignment of fluorinated proteins |
| title_full_unstemmed | Two-dimensional 19F–13C correlation NMR for 19F resonance assignment of fluorinated proteins |
| title_short | Two-dimensional 19F–13C correlation NMR for 19F resonance assignment of fluorinated proteins |
| title_sort | two dimensional 19f 13c correlation nmr for 19f resonance assignment of fluorinated proteins |
| url | https://hdl.handle.net/1721.1/136303 |
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