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X-ray Free Electron Laser Determination of Crystal Structures of Dark and Light States of a Reversibly Photoswitching Fluorescent Protein at Room Temperature.

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Title: X-ray Free Electron Laser Determination of Crystal Structures of Dark and Light States of a Reversibly Photoswitching Fluorescent Protein at Room Temperature.
Authors: Hutchison, CDM
Cordon-Preciado, V
Morgan, RML
Nakane, T
Ferreira, J
Dorlhiac, G
Sanchez-Gonzalez, A
Johnson, AS
Fitzpatrick, A
Fare, C
Marangos, JP
Yoon, CH
Hunter, MS
DePonte, DP
Boutet, S
Owada, S
Tanaka, R
Tono, K
Iwata, S
Van Thor, JJ
Item Type: Journal Article
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.
Issue Date: 7-Sep-2017
Date of Acceptance: 2-Sep-2017
URI: http://hdl.handle.net/10044/1/50526
DOI: https://dx.doi.org/10.3390/ijms18091918
ISSN: 1422-0067
Publisher: MDPI AG
Journal / Book Title: International Journal of Molecular Sciences
Volume: 18
Issue: 9
Copyright Statement: This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. (CC BY 4.0).
Keywords: LCLS
SACLA
SFX
Skylan-NS
XFEL
rsFP
0399 Other Chemical Sciences
0604 Genetics
0699 Other Biological Sciences
Chemical Physics
Publication Status: Published online
Conference Place: Switzerland
Article Number: 1918
Appears in Collections:Quantum Optics and Laser Science
Physics
Faculty of Natural Sciences