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

被引：12

作者：

Hutchison, Christopher D. M. ^{[1
]}

Cordon-Preciado, Violeta ^{[1
]}

Morgan, Rhodri M. L. ^{[2
]}

Nakane, Takanori ^{[3
]}

Ferreira, Josie ^{[1
]}

Dorlhiac, Gabriel ^{[1
]}

Sanchez-Gonzalez, Alvaro ^{[4
]}

Johnson, Allan S. ^{[4
]}

Fitzpatrick, Ann ^{[5
]}

Fare, Clyde ^{[1
]}

Marangos, Jon P. ^{[4
]}

Yoon, Chun Hong ^{[6
]}

Hunter, Mark S. ^{[6
]}

DePonte, Daniel P. ^{[6
]}

Boutet, Sebastien ^{[6
]}

Owada, Shigeki ^{[7
]}

Tanaka, Rie ^{[7
]}

Tono, Kensuke ^{[7
,8
]}

Iwata, So ^{[7
,9
]}

van Thor, Jasper J. ^{[1
]}

机构：

[1] Imperial Coll London, Mol Biophys, South Kensington Campus, London SW7 2AZ, England

[2] Imperial Coll London, Prot Crystallog Facil, Struct Biol Ctr, Dept Life Sci, Flowers Bldg, London SW7 2AZ, England

[3] Univ Tokyo, Grad Sch Sci, Dept Biol Sci, Bunkyo Ku, 2-11-16 Yayoi, Tokyo 1130032, Japan

[4] Imperial Coll, Blackett Lab, Quantum Opt & Laser Sci Grp, London SW7 2AZ, England

[5] Diamond Light Source Ltd, Diamond House,Harwell Sci & Innovat Campus, Didcot OX11 0DE, Oxon, England

[6] SLAC Natl Accelerator Lab, LCLS, 2575 Sand Hill Rd, Menlo Pk, CA 94025 USA

[7] RIKEN SPring 8 Ctr, 1-1-1 Kouto, Sayo, Hyogo 6795148, Japan

[8] Japan Synchrotron Radiat Res Inst, 1-1-1 Kouto, Sayo, Hyogo 6795198, Japan

[9] Kyoto Univ, Grad Sch Med, Dept Cell Biol, Sakyo Ku, Yoshidakonoe Cho, Kyoto 6068501, Japan

来源：

INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES | 2017年 / 18卷 / 09期

基金：

英国生物技术与生命科学研究理事会; 英国工程与自然科学研究理事会; 欧洲研究理事会; 英国科学技术设施理事会;

关键词：

XFEL; SFX; rsFP; Skylan-NS; SACLA; LCLS; ENERGY-TRANSFER; CRYSTALLOGRAPHY; REFINEMENT; PHOTOISOMERIZATION; DIFFRACTION; DRONPA; MODEL;

D O I：

10.3390/ijms18091918

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

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.

引用

页数：16

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