Spatiotemporal Resolution of Conformational Changes in Biomolecules by Combining Pulsed Electron-Electron Double Resonance Spectroscopy with Microsecond Freeze-Hyperquenching
| Autor(en): | Hett, Tobias Zbik, Tobias Mukherjee, Shatanik Matsuoka, Hideto Boenigk, Wolfgang Klose, Daniel Rouillon, Christophe Brenner, Norbert Peuker, Sebastian Klement, Reinhard Steinhoff, Heinz-Juergen Grubmueller, Helmut Seifert, Reinhard Schiemann, Olav Kaupp, U. Benjamin |
Stichwörter: | Chemistry; Chemistry, Multidisciplinary; DEER; DISTANCE MEASUREMENTS; DISTRIBUTIONS; EPR; KINETICS; MECHANOSENSITIVE CHANNEL; NUCLEOTIDE-BINDING DOMAIN; PROTEINS; RNA; STRUCTURAL BASIS | Erscheinungsdatum: | 2021 | Herausgeber: | AMER CHEMICAL SOC | Journal: | JOURNAL OF THE AMERICAN CHEMICAL SOCIETY | Volumen: | 143 | Ausgabe: | 18 | Startseite: | 6981 | Seitenende: | 6989 | Zusammenfassung: | The function of proteins is linked to their conformations that can be resolved with several high-resolution methods. However, only a few methods can provide the temporal order of intermediates and conformational changes, with each having its limitations. Here, we combine pulsed electron-electron double resonance spectroscopy with a microsecond freeze-hyperquenching setup to achieve spatiotemporal resolution in the angstrom range and lower microsecond time scale. We show that the conformational change of the C-alpha-helix in the cyclic nucleotide-binding domain of the Mesorhizobium loci potassium channel occurs within about 150 mu s and can be resolved with angstrom precision. Thus, this approach holds great promise for obtaining 4D landscapes of conformational changes in biomolecules. |
ISSN: | 00027863 | DOI: | 10.1021/jacs.1c01081 |
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