5OQA

XFEL structure of the off state of a reversibly photoswitching fluorescent protein determined using the GDVN injection method

Summary for 5OQA

• Entry DOI: 10.2210/pdb5oqa/pdb
• Descriptor: Green to red photoconvertible GFP-like protein EosFP (2 entities in total)
• Functional Keywords: xfel photoswitching, fluorescent protein
• Biological source: Lobophyllia hemprichii (Lobed brain coral)
• Total number of polymer chains: 1
• Total formula weight: 25724.27

Authors

Hutchison, C.D.M.,Cordon-Preciado, V.,Morgan, R.M.L.,van Thor, J.J. (deposition date: 2017-08-11, release date: 2017-12-06, Last modification date: 2024-11-13)

Primary citation

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

Experimental method

X-RAY DIFFRACTION (1.53 Å)