X-ray Free Electron Laser Determination of Crystal Structures of Dark and Light States of a Reversibly Photoswitching Fluorescent Protein at Room Temperature
| العنوان: | X-ray Free Electron Laser Determination of Crystal Structures of Dark and Light States of a Reversibly Photoswitching Fluorescent Protein at Room Temperature |
|---|---|
| المؤلفون: | Clyde Fare, Gabriel Dorlhiac, Mark S. Hunter, Sébastien Boutet, Rhodri M. L. Morgan, Jasper J. van Thor, Chun Hong Yoon, Daniel P. DePonte, So Iwata, Ann Fitzpatrick, Rie Tanaka, Takanori Nakane, C. Hutchison, Kensuke Tono, Shigeki Owada, Violeta Cordon-Preciado, Allan S. Johnson, Jon P. Marangos, Alvaro Sanchez-Gonzalez, Josie L. Ferreira |
| المصدر: | International Journal of Molecular Sciences International Journal of Molecular Sciences; Volume 18; Issue 9; Pages: 1918 International Journal of Molecular Sciences, Vol 18, Iss 9, p 1918 (2017) |
| بيانات النشر: | MDPI AG, 2017. |
| سنة النشر: | 2017 |
| مصطلحات موضوعية: | XFEL, SFX, rsFP, Skylan-NS, SACLA, LCLS, 0301 basic medicine, Protein Conformation, 0699 Other Biological Sciences, Analytical chemistry, Crystallography, X-Ray, Article, Catalysis, law.invention, lcsh:Chemistry, Inorganic Chemistry, Crystal, 03 medical and health sciences, law, 0399 Other Chemical Sciences, Physical and Theoretical Chemistry, lcsh:QH301-705.5, Molecular Biology, Spectroscopy, 0604 Genetics, Chemical Physics, Chemistry, Lasers, Organic Chemistry, Resolution (electron density), Temperature, General Medicine, Chromophore, Laser, Fluorescence, Computer Science Applications, Luminescent Proteins, 030104 developmental biology, lcsh:Biology (General), lcsh:QD1-999, Orthorhombic crystal system, Ground state |
| الوصف: | 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. |
| وصف الملف: | application/pdf |
| تدمد: | 1422-0067 |
| URL الوصول: | https://explore.openaire.eu/search/publication?articleId=doi_dedup___::bb9342ce459f27cb9fa826e0efa92dc9 https://doi.org/10.3390/ijms18091918 |
| حقوق: | OPEN |
| رقم الأكسشن: | edsair.doi.dedup.....bb9342ce459f27cb9fa826e0efa92dc9 |
| قاعدة البيانات: | OpenAIRE |
| تدمد: | 14220067 |
|---|