eLife (Mar 2021)
Time-resolved serial femtosecond crystallography reveals early structural changes in channelrhodopsin
- Kazumasa Oda,
- Takashi Nomura,
- Takanori Nakane,
- Keitaro Yamashita,
- Keiichi Inoue,
- Shota Ito,
- Johannes Vierock,
- Kunio Hirata,
- Andrés D Maturana,
- Kota Katayama,
- Tatsuya Ikuta,
- Itsuki Ishigami,
- Tamaki Izume,
- Rie Umeda,
- Ryuun Eguma,
- Satomi Oishi,
- Go Kasuya,
- Takafumi Kato,
- Tsukasa Kusakizako,
- Wataru Shihoya,
- Hiroto Shimada,
- Tomoyuki Takatsuji,
- Mizuki Takemoto,
- Reiya Taniguchi,
- Atsuhiro Tomita,
- Ryoki Nakamura,
- Masahiro Fukuda,
- Hirotake Miyauchi,
- Yongchan Lee,
- Eriko Nango,
- Rie Tanaka,
- Tomoyuki Tanaka,
- Michihiro Sugahara,
- Tetsunari Kimura,
- Tatsuro Shimamura,
- Takaaki Fujiwara,
- Yasuaki Yamanaka,
- Shigeki Owada,
- Yasumasa Joti,
- Kensuke Tono,
- Ryuichiro Ishitani,
- Shigehiko Hayashi,
- Hideki Kandori,
- Peter Hegemann,
- So Iwata,
- Minoru Kubo,
- Tomohiro Nishizawa,
- Osamu Nureki
Affiliations
- Kazumasa Oda
- ORCiD
- Department of Biological Sciences, Graduate School of Science, The University of Tokyo, Tokyo, Japan
- Takashi Nomura
- Graduate School of Life Science, University of Hyogo, Hyogo, Japan
- Takanori Nakane
- ORCiD
- Department of Biological Sciences, Graduate School of Science, The University of Tokyo, Tokyo, Japan
- Keitaro Yamashita
- Department of Biological Sciences, Graduate School of Science, The University of Tokyo, Tokyo, Japan
- Keiichi Inoue
- ORCiD
- Graduate School of Engineering, Nagoya Institute of Technology, Nagoya, Japan
- Shota Ito
- Graduate School of Engineering, Nagoya Institute of Technology, Nagoya, Japan
- Johannes Vierock
- ORCiD
- Institute of Biology, Experimental Biophysics, Humboldt-Universität zu Berlin, Berlin, Germany
- Kunio Hirata
- RIKEN SPring-8 Center, Hyogo, Japan; Precursory Research for Embryonic Science and Technology (PRESTO), Japan Science and Technology Agency, Kawaguchi, Japan
- Andrés D Maturana
- Department of Bioengineering Sciences, Graduate School of Bioagricultural Sciences, Nagoya University, Nagoya, Japan
- Kota Katayama
- Graduate School of Engineering, Nagoya Institute of Technology, Nagoya, Japan
- Tatsuya Ikuta
- Department of Biological Sciences, Graduate School of Science, The University of Tokyo, Tokyo, Japan
- Itsuki Ishigami
- Department of Biological Sciences, Graduate School of Science, The University of Tokyo, Tokyo, Japan
- Tamaki Izume
- Department of Biological Sciences, Graduate School of Science, The University of Tokyo, Tokyo, Japan
- Rie Umeda
- Department of Biological Sciences, Graduate School of Science, The University of Tokyo, Tokyo, Japan
- Ryuun Eguma
- Department of Biological Sciences, Graduate School of Science, The University of Tokyo, Tokyo, Japan
- Satomi Oishi
- Department of Biological Sciences, Graduate School of Science, The University of Tokyo, Tokyo, Japan
- Go Kasuya
- ORCiD
- Department of Biological Sciences, Graduate School of Science, The University of Tokyo, Tokyo, Japan
- Takafumi Kato
- Department of Biological Sciences, Graduate School of Science, The University of Tokyo, Tokyo, Japan
- Tsukasa Kusakizako
- Department of Biological Sciences, Graduate School of Science, The University of Tokyo, Tokyo, Japan
- Wataru Shihoya
- Department of Biological Sciences, Graduate School of Science, The University of Tokyo, Tokyo, Japan
- Hiroto Shimada
- Department of Biological Sciences, Graduate School of Science, The University of Tokyo, Tokyo, Japan
- Tomoyuki Takatsuji
- Department of Biological Sciences, Graduate School of Science, The University of Tokyo, Tokyo, Japan
- Mizuki Takemoto
- ORCiD
- Department of Biological Sciences, Graduate School of Science, The University of Tokyo, Tokyo, Japan
- Reiya Taniguchi
- Department of Biological Sciences, Graduate School of Science, The University of Tokyo, Tokyo, Japan
- Atsuhiro Tomita
- Department of Biological Sciences, Graduate School of Science, The University of Tokyo, Tokyo, Japan
- Ryoki Nakamura
- Department of Biological Sciences, Graduate School of Science, The University of Tokyo, Tokyo, Japan
- Masahiro Fukuda
- Department of Biological Sciences, Graduate School of Science, The University of Tokyo, Tokyo, Japan
- Hirotake Miyauchi
- Department of Biological Sciences, Graduate School of Science, The University of Tokyo, Tokyo, Japan
- Yongchan Lee
- Department of Biological Sciences, Graduate School of Science, The University of Tokyo, Tokyo, Japan
- Eriko Nango
- ORCiD
- RIKEN SPring-8 Center, Hyogo, Japan; Department of Cell Biology, Graduate School of Medicine, Kyoto University, Kyoto, Japan
- Rie Tanaka
- RIKEN SPring-8 Center, Hyogo, Japan; Department of Cell Biology, Graduate School of Medicine, Kyoto University, Kyoto, Japan
- Tomoyuki Tanaka
- RIKEN SPring-8 Center, Hyogo, Japan; Department of Cell Biology, Graduate School of Medicine, Kyoto University, Kyoto, Japan
- Michihiro Sugahara
- RIKEN SPring-8 Center, Hyogo, Japan
- Tetsunari Kimura
- Department of Chemistry, Graduate School of Science, Kobe University, Kobe, Japan
- Tatsuro Shimamura
- Department of Cell Biology, Graduate School of Medicine, Kyoto University, Kyoto, Japan
- Takaaki Fujiwara
- Department of Cell Biology, Graduate School of Medicine, Kyoto University, Kyoto, Japan
- Yasuaki Yamanaka
- Department of Cell Biology, Graduate School of Medicine, Kyoto University, Kyoto, Japan
- Shigeki Owada
- RIKEN SPring-8 Center, Hyogo, Japan; Japan Synchrotron Radiation Research Institute, Hyogo, Japan
- Yasumasa Joti
- RIKEN SPring-8 Center, Hyogo, Japan; Japan Synchrotron Radiation Research Institute, Hyogo, Japan
- Kensuke Tono
- RIKEN SPring-8 Center, Hyogo, Japan; Japan Synchrotron Radiation Research Institute, Hyogo, Japan
- Ryuichiro Ishitani
- ORCiD
- Department of Biological Sciences, Graduate School of Science, The University of Tokyo, Tokyo, Japan
- Shigehiko Hayashi
- Department of Chemistry, Graduate School of Science, Kyoto University, Kyoto, Japan
- Hideki Kandori
- Graduate School of Engineering, Nagoya Institute of Technology, Nagoya, Japan
- Peter Hegemann
- ORCiD
- Institute of Biology, Experimental Biophysics, Humboldt-Universität zu Berlin, Berlin, Germany
- So Iwata
- RIKEN SPring-8 Center, Hyogo, Japan; Department of Cell Biology, Graduate School of Medicine, Kyoto University, Kyoto, Japan
- Minoru Kubo
- Graduate School of Life Science, University of Hyogo, Hyogo, Japan
- Tomohiro Nishizawa
- ORCiD
- Department of Biological Sciences, Graduate School of Science, The University of Tokyo, Tokyo, Japan; Precursory Research for Embryonic Science and Technology (PRESTO), Japan Science and Technology Agency, Kawaguchi, Japan
- Osamu Nureki
- ORCiD
- Department of Biological Sciences, Graduate School of Science, The University of Tokyo, Tokyo, Japan
- DOI
- https://doi.org/10.7554/eLife.62389
- Journal volume & issue
-
Vol. 10
Abstract
Channelrhodopsins (ChRs) are microbial light-gated ion channels utilized in optogenetics to control neural activity with light . Light absorption causes retinal chromophore isomerization and subsequent protein conformational changes visualized as optically distinguished intermediates, coupled with channel opening and closing. However, the detailed molecular events underlying channel gating remain unknown. We performed time-resolved serial femtosecond crystallographic analyses of ChR by using an X-ray free electron laser, which revealed conformational changes following photoactivation. The isomerized retinal adopts a twisted conformation and shifts toward the putative internal proton donor residues, consequently inducing an outward shift of TM3, as well as a local deformation in TM7. These early conformational changes in the pore-forming helices should be the triggers that lead to opening of the ion conducting pore.
Keywords
