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A photoproduct of DXCF cyanobacteriochromes without reversible Cys ligation is destabilized by rotating ring twist of the chromophore

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Abstract

Cyanobacteriochrome photoreceptors (CBCRs) ligate linear tetrapyrrole chromophores via their first (canonical) Cys residue and show reversible photoconversion triggered by light-dependent Z/E isomeri-zation of the chromophore. Among the huge repertoire of CBCRs, DXCF CBCRs contain a second Cys residue within the highly conserved Asp-Xaa-Cys-Phe (DXCF) motif. In the typical receptors, the second Cys covalently attaches to the 15Z-chromophore in the dark state and detaches from the 15E-chromophore in the photoproduct state, whereas atypical ones that lack reversible ligation activity show redshifted absorption in the dark state due to a more extended π -conjugated system. Moreover, some DXCF CBCRs show blue-shifted absorption in the photoproduct state due to the twisted geometry of the rotating ring. During the process of rational color tuning of a certain DXCF CBCR, we unexpectedly found that twisted photoproducts of some variant molecules showed dark reversion to the dark state, which prompted us to hypothesize that the photoproduct is destabilized by the twisted geometry of the rotating ring. In this study, we comprehensively examined the photoproduct stability of the twisted and relaxed molecules derived from the same CBCR scaffolds under dark conditions. In the DXCF CBCRs lacking reversible ligation activity, the twisted photoproducts showed faster dark reversion than the relaxed ones, supporting our hypothesis. By contrast, in the DXCF CBCRs exhibiting reversible ligation activity, the twisted photoproducts showed no detectable photoconversion. Reversible Cys adduct formation thus results in drastic rearrangement of the protein–chromophore interaction in the photoproduct state, which would contribute to the previously unknown photoproduct stability.

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Authors and Affiliations

  1. Department of Biological Sciences, Faculty of Science, Shizuoka University, 836 Ohya, Suruga, Shizuoka, 422-8529, Japan

    Keiji Fushimi, Takumi Matsunaga & Rei Narikawa

  2. Core Research for Evolutional Science and Technology, Japan Science and Technology Agency, 4-1-8 Honcho, Kawaguchi, Saitama, 332-0012, Japan

    Keiji Fushimi & Rei Narikawa

  3. Research Institute of Green Science and Technology, Shizuoka University, 836 Ohya, Suruga, Shizuoka, 422-8529, Japan

    Rei Narikawa

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  1. Keiji Fushimi
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  2. Takumi Matsunaga
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Correspondence to Rei Narikawa.

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Electronic supplementary information (ESI) available. See DOI: 10.1039/ d0pp00208a

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Fushimi, K., Matsunaga, T. & Narikawa, R. A photoproduct of DXCF cyanobacteriochromes without reversible Cys ligation is destabilized by rotating ring twist of the chromophore. Photochem Photobiol Sci 19, 1289–1299 (2020). https://doi.org/10.1039/d0pp00208a

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