8A2V

Room temperature structure of the ground state of AtPhot2LOV2 in space group P43212

8A2V の概要

• エントリーDOI: 10.2210/pdb8a2v/pdb
• 分子名称: Phototropin-2, FLAVIN MONONUCLEOTIDE, 2-(N-MORPHOLINO)-ETHANESULFONIC ACID, ... (4 entities in total)
• 機能のキーワード: lov domain, plant protein
• 由来する生物種: Arabidopsis thaliana (thale cress)
• タンパク質・核酸の鎖数: 1
• 化学式量合計: 15659.49

構造登録者

Engilberge, S.,Caramello, N.,Royant, A. (登録日: 2022-06-06, 公開日: 2023-03-29, 最終更新日: 2024-02-07)

主引用文献

Aumonier, S.,Engilberge, S.,Caramello, N.,von Stetten, D.,Gotthard, G.,Leonard, G.A.,Mueller-Dieckmann, C.,Royant, A.
Slow protein dynamics probed by time-resolved oscillation crystallography at room temperature.
Iucrj, 9:756-767, 2022

PubMed Abstract: 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 , 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 UV-Vis absorption spectroscopy, this RT-MX approach allowed the authors to follow the relaxation of the photoadduct, a thio-ether 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.

PubMed: 36381146
DOI: 10.1107/S2052252522009150

実験手法

X-RAY DIFFRACTION (1.59 Å)