We found a match
Your institution may have rights to this item. Sign in to continue.
- 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, Christopher D. M.; Cordon-Preciado, Violeta; Ferreira, Josie; Dorlhiac, Gabriel; Fare, Clyde; van Thor, Jasper J.; Morgan, Rhodri M. L.; Takanori Nakane; Sanchez-Gonzalez, Alvaro; Johnson, Allan S.; Marangos, Jon P.; Fitzpatrick, Ann; Chun Hong Yoon; Hunter, Mark S.; DePonte, Daniel P.; Boutet, Sébastien; Shigeki Owada; Rie Tanaka; Kensuke Tono; So Iwata
- 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.
- Subjects
X-rays; CRYSTAL structure; FLUORESCENT proteins; PHOTOELECTRICITY; CHROMOPHORES
- Publication
International Journal of Molecular Sciences, 2017, Vol 18, Issue 9, p1918
- ISSN
1661-6596
- Publication type
Article
- DOI
10.3390/ijms18091918