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Atomistic design of microbial opsin-based blue-shifted optogenetics tools

Author

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  • Hideaki E. Kato

    (Graduate School of Science, The University of Tokyo
    Present address: Department of Molecular and Cellular Physiology, Stanford University School of Medicine, Stanford, California 94305, USA)

  • Motoshi Kamiya

    (Graduate School of Science, Kyoto University)

  • Seiya Sugo

    (Graduate School of Science, Kyoto University)

  • Jumpei Ito

    (Graduate School of Bioagricultural Sciences, Nagoya University)

  • Reiya Taniguchi

    (Graduate School of Science, The University of Tokyo)

  • Ayaka Orito

    (Graduate School of Bioagricultural Sciences, Nagoya University)

  • Kunio Hirata

    (RIKEN SPring-8 Center)

  • Ayumu Inutsuka

    (Research Institute of Environmental Medicine, Nagoya University)

  • Akihiro Yamanaka

    (Research Institute of Environmental Medicine, Nagoya University)

  • Andrés D. Maturana

    (Graduate School of Bioagricultural Sciences, Nagoya University)

  • Ryuichiro Ishitani

    (Graduate School of Science, The University of Tokyo)

  • Yuki Sudo

    (Graduate School of Medicine, Dentistry, and Pharmaceutical Sciences, Okayama University)

  • Shigehiko Hayashi

    (Graduate School of Science, Kyoto University)

  • Osamu Nureki

    (Graduate School of Science, The University of Tokyo)

Abstract

Microbial opsins with a bound chromophore function as photosensitive ion transporters and have been employed in optogenetics for the optical control of neuronal activity. Molecular engineering has been utilized to create colour variants for the functional augmentation of optogenetics tools, but was limited by the complexity of the protein–chromophore interactions. Here we report the development of blue-shifted colour variants by rational design at atomic resolution, achieved through accurate hybrid molecular simulations, electrophysiology and X-ray crystallography. The molecular simulation models and the crystal structure reveal the precisely designed conformational changes of the chromophore induced by combinatory mutations that shrink its π-conjugated system which, together with electrostatic tuning, produce large blue shifts of the absorption spectra by maximally 100 nm, while maintaining photosensitive ion transport activities. The design principle we elaborate is applicable to other microbial opsins, and clarifies the underlying molecular mechanism of the blue-shifted action spectra of microbial opsins recently isolated from natural sources.

Suggested Citation

  • Hideaki E. Kato & Motoshi Kamiya & Seiya Sugo & Jumpei Ito & Reiya Taniguchi & Ayaka Orito & Kunio Hirata & Ayumu Inutsuka & Akihiro Yamanaka & Andrés D. Maturana & Ryuichiro Ishitani & Yuki Sudo & Sh, 2015. "Atomistic design of microbial opsin-based blue-shifted optogenetics tools," Nature Communications, Nature, vol. 6(1), pages 1-10, November.
  • Handle: RePEc:nat:natcom:v:6:y:2015:i:1:d:10.1038_ncomms8177
    DOI: 10.1038/ncomms8177
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    Cited by:

    1. Kyle Tucker & Savitha Sridharan & Hillel Adesnik & Stephen G. Brohawn, 2022. "Cryo-EM structures of the channelrhodopsin ChRmine in lipid nanodiscs," Nature Communications, Nature, vol. 13(1), pages 1-12, December.
    2. R. Astashkin & K. Kovalev & S. Bukhdruker & S. Vaganova & A. Kuzmin & A. Alekseev & T. Balandin & D. Zabelskii & I. Gushchin & A. Royant & D. Volkov & G. Bourenkov & E. Koonin & M. Engelhard & E. Bamb, 2022. "Structural insights into light-driven anion pumping in cyanobacteria," Nature Communications, Nature, vol. 13(1), pages 1-13, December.

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