Optogenetic cytosol acidification of mammalian cells using an inward proton-pumping rhodopsin.

International journal of biological macromolecules 2023 Jul 01

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Ion gradients are a universal form of energy, information storage and conversion in living cells. Advances in optogenetics inspire the development of novel tools towards control of different cellular processes with light. Rhodopsins are perspective tools for optogenetic manipulation of ion gradients in cells and subcellular compartments, controlling pH of the cytosol and intracellular organelles. The key step of the development of new optogenetic tools is evaluation of their efficiency. Here, we used a high-throughput quantitative method for comparing efficiency of proton-pumping rhodopsins in Escherichia coli cells. This approach allowed us to show that an inward proton pump xenorhodopsin from Nanosalina sp. (NsXeR) is a powerful tool for optogenetic control of pH of mammalian subcellular compartments. Further, we demonstrate that NsXeR can be used for fast optogenetic acidification of the cytosol of mammalian cells. This is the first evidence of optogenetic cytosol acidification by an inward proton pump at physiological pH values. Our approach offers unique opportunities to study cellular metabolism at normal and pathological conditions and might help to understand the role of pH dysregulation in cellular dysfunctions. Copyright © 2023 Elsevier B.V. All rights reserved.

Citation

A Vlasova, A Polyakova, A Gromova, S Dolotova, S Bukhalovich, D Bagaeva, N Bondarev, F Tsybrov, K Kovalev, A Mikhailov, D Sidorov, A Bogorodskiy, N Ilyinsky, A Kuklin, A Vlasov, V Borshchevskiy, V Ivanovich. Optogenetic cytosol acidification of mammalian cells using an inward proton-pumping rhodopsin. International journal of biological macromolecules. 2023 Jul 01;242(Pt 3):124949