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Fusion Proteins Containing Proteorhodopsin from Exiguobacterium sibiricum

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Abstract

Objective: To study the transport activity of bacterial rhodopsins, proteoliposomes with the same orientation of the protein molecules should be obtained. Methods: The genes of fusion proteins containing Exiguobacterium sibiricum proteorhodopsin (ESR) and various N-terminal soluble domains have been constructed. Results and Discussion: Effective synthesis in Escherichia coli cells was observed only in the case of fusion with chaperone Caf1M and maltose-binding protein MBP expressed as precursors with their own signal sequences. The study of the isolated MBP-ESR protein in micelles and proteoliposomes demonstrated formation and decay of the main photocycle intermediates at pH > 8. The photoelectric response of the fusion proteins Caf-ESR and MBP-ESR is comparable in amplitude to the wild-type ESR response, indicating their homogeneous orientation in the proteoliposome membrane. Conclusions: The obtained constructs can be used to create bacterial expression systems for various retinal proteins, ensuring their uniform incorporation into proteoliposomes.

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DATA AVAILABILITY

The data that support the findings of this study are available from the corresponding author upon reasonable request.

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Funding

This work was supported by the Russian Federation Ministry of Science and Higher Education grant (agreement no. 075-15-2020-795, project ID RF-190220X0027).

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Authors and Affiliations

  1. Shemyakin–Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences, 117997, Moscow, Russia

    L. E. Petrovskaya, E. A. Kryukova, D. A. Dolgikh & M. P. Kirpichnikov

  2. Moscow Institute of Physics and Technology, 141701, Dolgoprudny, Russia

    L. E. Petrovskaya

  3. Emanuel Institute of Biochemical Physics, 119334, Moscow, Russia

    E. A. Kryukova & D. A. Dolgikh

  4. Faculty of Biology, Lomonosov Moscow State University, 119234, Moscow, Russia

    V. A. Bolshakov, E. P. Lukashev, D. A. Dolgikh & M. P. Kirpichnikov

  5. Belozersky Institute of Physico-Chemical Biology, Lomonosov Moscow State University, 119992, Moscow, Russia

    S. A. Siletsky, M. D. Mamedov & R. V. Sudakov

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  1. L. E. Petrovskaya
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  2. E. A. Kryukova
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Petrovskaya, L.E., Kryukova, E.A., Bolshakov, V.A. et al. Fusion Proteins Containing Proteorhodopsin from Exiguobacterium sibiricum. Russ J Bioorg Chem 50, 741–752 (2024). https://doi.org/10.1134/S1068162024030063

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