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Nat Commun. 2014 Dec 15;5:5810. doi: 10.1038/ncomms6810.

Synthetic retinal analogues modify the spectral and kinetic characteristics of microbial rhodopsin optogenetic tools.

Author information

1
1] Buchmann Institute for Molecular Life Sciences (BMLS), Goethe-University, Max-von-Laue-Straße 15, 60438 Frankfurt, Germany [2] Institute of Biochemistry, Goethe-University, Max-von-Laue-Straße 9, 60438 Frankfurt, Germany.
2
Institute for Biology-Experimental Biophysics, Humboldt-Universität zu Berlin, Invalidenstraße 42, 10115 Berlin, Germany.
3
Department of Molecular Neurobiology of Behavior, Georg-August-Universität Göttingen, Julia-Lermontowa-Weg 3, 37077 Göttingen, Germany.
4
Endotherm, Science-Park II, 66123 Saarbrücken, Germany.
5
Department of Chemistry, Ludwig-Maximilians-Universität München, Butenandtstraße 5-13, 81377 München, Germany.
6
Institute of Biochemistry, Goethe-University, Max-von-Laue-Straße 9, 60438 Frankfurt, Germany.
7
Department of Organic Chemistry, The Weizmann Institute of Science, Rehovot 76100, Israel.
8
1] Buchmann Institute for Molecular Life Sciences (BMLS), Goethe-University, Max-von-Laue-Straße 15, 60438 Frankfurt, Germany [2] Institute of Biochemistry, Goethe-University, Max-von-Laue-Straße 9, 60438 Frankfurt, Germany [3] Cluster of Excellence Frankfurt Macromolecular Complexes (CEF-MC), Goethe University, Max-von-Laue Straße 15 60438, Frankfurt, Germany.

Abstract

Optogenetic tools have become indispensable in neuroscience to stimulate or inhibit excitable cells by light. Channelrhodopsin-2 (ChR2) variants have been established by mutating the opsin backbone or by mining related algal genomes. As an alternative strategy, we surveyed synthetic retinal analogues combined with microbial rhodopsins for functional and spectral properties, capitalizing on assays in C. elegans, HEK cells and larval Drosophila. Compared with all-trans retinal (ATR), Dimethylamino-retinal (DMAR) shifts the action spectra maxima of ChR2 variants H134R and H134R/T159C from 480 to 520 nm. Moreover, DMAR decelerates the photocycle of ChR2(H134R) and (H134R/T159C), thereby reducing the light intensity required for persistent channel activation. In hyperpolarizing archaerhodopsin-3 and Mac, naphthyl-retinal and thiophene-retinal support activity alike ATR, yet at altered peak wavelengths. Our experiments enable applications of retinal analogues in colour tuning and altering photocycle characteristics of optogenetic tools, thereby increasing the operational light sensitivity of existing cell lines or transgenic animals.

PMID:
25503804
DOI:
10.1038/ncomms6810
[Indexed for MEDLINE]

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