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Photocurrent attenuation by a single polar-to-nonpolar point mutation of channelrhodopsin-2

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Abstract

Channelrhodopsin-2 (ChR2), one of the algal light-gatedcation channel rhodopsins, contains five peculiar glutamic acid residues in the N-terminal region corresponding to the second to third transmembrane helices. Here we made systematic mutations of these polar amino acid residues of ChR2 into nonpolar alanine, and evaluated their photocurrent properties. Amongst them, the photocurrent generated by the E97A mutation, ChR2(E97A), was much smaller than expected from its expression. The ChR2(E97A) photocurrent was similar to wild-type ChR2 in the kinetic profiles, the reversal potential and the dependency to the light power density. Our results suggest that the residue E97 is one of the molecular determinants involved in the ion flux regulation.

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Authors and Affiliations

  1. Department of Physiology and Pharmacology, Tohoku University Graduate School of Medicine, Sendai, 980-8575, Japan

    Yuka Sugiyama & Hiromu Yawo

  2. Tohoku University Basic and Translational Research Center for Global Brain Science, Sendai, 980-8575, Japan

    Yuka Sugiyama, Hongxia Wang, Takuya Hikima & Hiromu Yawo

  3. Department of Developmental Biology and Neuroscience, Tohoku University Graduate School of Life Sciences, 2-1 Seiryo-machi, Aoba-ku, Sendai, 980-8575, Japan

    Hongxia Wang, Takuya Hikima, Toru Ishizuka & Hiromu Yawo

  4. Faculty of Pharmaceutical Sciences, Toho University, Chiba, 274-8510, Japan

    Minami Sato, Jun Kuroda & Tetsuo Takahashi

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  1. Yuka Sugiyama
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Correspondence to Hiromu Yawo.

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Electronic supplementary information (ESI) available. See DOI: 10.1039/b815762f

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Sugiyama, Y., Wang, H., Hikima, T. et al. Photocurrent attenuation by a single polar-to-nonpolar point mutation of channelrhodopsin-2. Photochem Photobiol Sci 8, 328–336 (2009). https://doi.org/10.1039/b815762f

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  • DOI: https://doi.org/10.1039/b815762f

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