دورية أكاديمية
Slow protein dynamics probed by time-resolved oscillation crystallography at room temperature
| العنوان: | Slow protein dynamics probed by time-resolved oscillation crystallography at room temperature |
|---|---|
| المؤلفون: | Sylvain Aumonier, Sylvain Engilberge, Nicolas Caramello, David von Stetten, Guillaume Gotthard, Gordon A. Leonard, Christoph Mueller-Dieckmann, Antoine Royant |
| المصدر: | IUCrJ, Vol 9, Iss 6, Pp 756-767 (2022) |
| بيانات النشر: | International Union of Crystallography, 2022. |
| سنة النشر: | 2022 |
| المجموعة: | LCC:Crystallography |
| مصطلحات موضوعية: | room-temperature crystallography, time-resolved crystallography, protein dynamics, lov2 domain of phototropin 2, Crystallography, QD901-999 |
| الوصف: | The development of serial crystallography over the last decade at XFELs and synchrotrons has produced a renaissance in room-temperature macromolecular crystallography (RT-MX), and fostered many technical and methodological breakthroughs designed to study phenomena occurring in proteins on the picosecond-to-second timescale. However, there are components of protein dynamics that occur in much slower regimes, of which the study could readily benefit from state-of-the-art RT-MX. Here, the room-temperature structural study of the relaxation of a reaction intermediate at a synchrotron, exploiting a handful of single crystals, is described. The intermediate in question is formed in microseconds during the photoreaction of the LOV2 domain of phototropin 2 from Arabidopsis thaliana, which then decays in minutes. This work monitored its relaxation in the dark using a fast-readout EIGER X 4M detector to record several complete oscillation X-ray diffraction datasets, each of 1.2 s total exposure time, at different time points in the relaxation process. Coupled with in crystallo UV–Vis absorption spectroscopy, this RT-MX approach allowed the authors to follow the relaxation of the photoadduct, a thioether covalent bond between the chromophore and a cysteine residue. Unexpectedly, the return of the chromophore to its spectroscopic ground state is followed by medium-scale protein rearrangements that trigger a crystal phase transition and hinder the full recovery of the structural ground state of the protein. In addition to suggesting a hitherto unexpected role of a conserved tryptophan residue in the regulation of the photocycle of LOV2, this work provides a basis for performing routine time-resolved protein crystallography experiments at synchrotrons for phenomena occurring on the second-to-hour timescale. |
| نوع الوثيقة: | article |
| وصف الملف: | electronic resource |
| اللغة: | English |
| تدمد: | 2052-2525 20522525 |
| العلاقة: | http://scripts.iucr.org/cgi-bin/paper?S2052252522009150Test; https://doaj.org/toc/2052-2525Test |
| DOI: | 10.1107/S2052252522009150 |
| الوصول الحر: | https://doaj.org/article/1eb6c26f67784312934f75f9211900e9Test |
| رقم الانضمام: | edsdoj.1eb6c26f67784312934f75f9211900e9 |
| قاعدة البيانات: | Directory of Open Access Journals |
Full Text Finder | تدمد: | 20522525 |
|---|---|
| DOI: | 10.1107/S2052252522009150 |