5OOZ

XFEL structure of the on state of a reversibly photoswitching fluorescent protein determined using the grease injection method

5OOZ の概要

• エントリーDOI: 10.2210/pdb5ooz/pdb
• 分子名称: Green to red photoconvertible GFP-like protein EosFP (2 entities in total)
• 機能のキーワード: xfel photoswitching, fluorescent protein
• 由来する生物種: Lobophyllia hemprichii (Lobed brain coral)
• タンパク質・核酸の鎖数: 1
• 化学式量合計: 25725.22

構造登録者

Hutchison, C.D.M.,Cordon-Preciado, V.,Morgan, R.M.L.,van Thor, J.J. (登録日: 2017-08-09, 公開日: 2017-12-06, 最終更新日: 2024-10-23)

主引用文献

Hutchison, C.D.M.,Cordon-Preciado, V.,Morgan, R.M.L.,Nakane, T.,Ferreira, J.,Dorlhiac, G.,Sanchez-Gonzalez, A.,Johnson, A.S.,Fitzpatrick, A.,Fare, C.,Marangos, J.P.,Yoon, C.H.,Hunter, M.S.,DePonte, D.P.,Boutet, S.,Owada, S.,Tanaka, R.,Tono, K.,Iwata, S.,van Thor, J.J.
X-ray Free Electron Laser Determination of Crystal Structures of Dark and Light States of a Reversibly Photoswitching Fluorescent Protein at Room Temperature.
Int J Mol Sci, 18:-, 2017

PubMed Abstract: The photochromic fluorescent protein Skylan-NS (Nonlinear Structured illumination variant mEos3.1H62L) is a reversibly photoswitchable fluorescent protein which has an unilluminated/ground state with an anionic and cis chromophore conformation and high fluorescence quantum yield. Photo-conversion with illumination at 515 nm generates a meta-stable intermediate with neutral trans-chromophore structure that has a 4 h lifetime. We present X-ray crystal structures of the cis (on) state at 1.9 Angstrom resolution and the trans (off) state at a limiting resolution of 1.55 Angstrom from serial femtosecond crystallography experiments conducted at SPring-8 Angstrom Compact Free Electron Laser (SACLA) at 7.0 keV and 10.5 keV, and at Linac Coherent Light Source (LCLS) at 9.5 keV. We present a comparison of the data reduction and structure determination statistics for the two facilities which differ in flux, beam characteristics and detector technologies. Furthermore, a comparison of droplet on demand, grease injection and Gas Dynamic Virtual Nozzle (GDVN) injection shows no significant differences in limiting resolution. The photoconversion of the on- to the off-state includes both internal and surface exposed protein structural changes, occurring in regions that lack crystal contacts in the orthorhombic crystal form.

PubMed: 28880248
DOI: 10.3390/ijms18091918

実験手法

X-RAY DIFFRACTION (1.92 Å)