한빛사논문
Ji-Hye Yuna,1, Xuanxuan Lib,c,1, Jianing Yued, Jae-Hyun Parka, Zeyu Jina, Chufeng Lie, Hao Hue, Yingchen Shib,c, Suraj Pandeyf, Sergio Carbajog, Sébastien Boutetg, Mark S. Hunterg, Mengning Liangg, Raymond G. Sierrag, Thomas J. Laneg, Liang Zhoud, Uwe Weierstalle, Nadia A. Zatsepine,h, Mio Ohkii, Jeremy R. H. Tamei, Sam-Yong Parki, John C. H. Spencee, Wenkai Zhangd, Marius Schmidtf,2, Weontae Leea,2, and Haiguang Liub,d,2
aDepartment of Biochemistry, College of Life Sciences and Biotechnology, Yonsei University, Seodaemun-gu, 120-749 Seoul, South Korea; bComplex Systems Division, Beijing Computational Science Research Center, Haidian, 100193 Beijing, People’s Republic of China; cDepartment of Engineering Physics, Tsinghua University, 100086 Beijing, People’s Republic of China; dDepartment of Physics, Beijing Normal University, Haidian, 100875 Beijing, People’s Republic of China; eDepartment of Physics, Arizona State University, Tempe, AZ 85287; fPhysics Department, University of Wisconsin, Milwaukee, Milwaukee, WI 53201; gLinac Coherent Light Source, Stanford Linear Accelerator Center National Accelerator Laboratory, Menlo Park, CA 94025; hDepartment of Chemistry and Physics, Australian Research Council Centre of Excellence in Advanced Molecular Imaging, La Trobe Institute for Molecular Science, La Trobe University, Melbourne, VIC 3086, Australia; and iDrug Design Laboratory, Graduate School of Medical Life Science, Yokohama City University, 230-0045 Yokohama, Japan
1J.-H.Y. and X.L. contributed equally to this work.
2To whom correspondence may be addressed.
Abstract
Chloride ion–pumping rhodopsin (ClR) in some marine bacteria utilizes light energy to actively transport Cl− into cells. How the ClR initiates the transport is elusive. Here, we show the dynamics of ion transport observed with time-resolved serial femtosecond (fs) crystallography using the Linac Coherent Light Source. X-ray pulses captured structural changes in ClR upon flash illumination with a 550 nm fs-pumping laser. High-resolution structures for five time points (dark to 100 ps after flashing) reveal complex and coordinated dynamics comprising retinal isomerization, water molecule rearrangement, and conformational changes of various residues. Combining data from time-resolved spectroscopy experiments and molecular dynamics simulations, this study reveals that the chloride ion close to the Schiff base undergoes a dissociation–diffusion process upon light-triggered retinal isomerization.
time-resolved crystallography, light-driven chloride-pumping rhodopsin, X-ray free-electron laser, serial femtosecond crystallography
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