The Desensitized Channelrhodopsin-2 Photointermediate Contains 13-cis,15-syn retinal schiff base
The Desensitized Channelrhodopsin-2 Photointermediate Contains 13-cis,15-syn retinal schiff base
Channelrhodopsin-2 (ChR2) is a light-gated cation channel and was used to lay the foundations of optogenetics. Its dark state X-ray structure has been determined in 2017 for the wild-type, which is the prototype for all other ChR variants. However, the mechanistic understanding of the channel function is still incomplete in terms of structural changes after photon absorption by the retinal chromophore and in the framework of functional models. Hence, detailed information needs to be collected on the dark state as well as on the different photointermediates. For ChR2 detailed knowledge on the chromophore configuration in the different states is still missing and a consensus has not been achieved. Using DNP-enhanced solid-state MAS NMR spectroscopy on proteoliposome samples, we unambiguously determined the chromophore configuration in the desensitized state, and we show that this state occurs towards the end of the photocycle.
Photocycle, channelrhodopsin, dynamic nuclear polarization, membrane proteins, solid-state nmr spectroscopy
16442-16447
Becker-Baldus, Johanna
6563d44b-d7aa-4e93-b7c2-57b95f8a0f1d
Leeder, Alexander J
82f20bdd-9d92-47b8-8720-6f5c66db4be7
Brown, Lynda J.
75aa95fa-5d27-46a7-9dbe-0f465a664f5b
Brown, Richard C.D.
21ce697a-7c3a-480e-919f-429a3d8550f5
Bamann, Christian
332a26c0-befa-43b0-a3b6-efa344cf644c
Glaubitz, Clemens
99f5e847-e6fd-4783-bc60-054bf0e15661
19 July 2021
Becker-Baldus, Johanna
6563d44b-d7aa-4e93-b7c2-57b95f8a0f1d
Leeder, Alexander J
82f20bdd-9d92-47b8-8720-6f5c66db4be7
Brown, Lynda J.
75aa95fa-5d27-46a7-9dbe-0f465a664f5b
Brown, Richard C.D.
21ce697a-7c3a-480e-919f-429a3d8550f5
Bamann, Christian
332a26c0-befa-43b0-a3b6-efa344cf644c
Glaubitz, Clemens
99f5e847-e6fd-4783-bc60-054bf0e15661
Becker-Baldus, Johanna, Leeder, Alexander J, Brown, Lynda J., Brown, Richard C.D., Bamann, Christian and Glaubitz, Clemens
(2021)
The Desensitized Channelrhodopsin-2 Photointermediate Contains 13-cis,15-syn retinal schiff base.
Angewandte Chemie International Edition, 60 (30), .
(doi:10.1002/anie.202015797).
Abstract
Channelrhodopsin-2 (ChR2) is a light-gated cation channel and was used to lay the foundations of optogenetics. Its dark state X-ray structure has been determined in 2017 for the wild-type, which is the prototype for all other ChR variants. However, the mechanistic understanding of the channel function is still incomplete in terms of structural changes after photon absorption by the retinal chromophore and in the framework of functional models. Hence, detailed information needs to be collected on the dark state as well as on the different photointermediates. For ChR2 detailed knowledge on the chromophore configuration in the different states is still missing and a consensus has not been achieved. Using DNP-enhanced solid-state MAS NMR spectroscopy on proteoliposome samples, we unambiguously determined the chromophore configuration in the desensitized state, and we show that this state occurs towards the end of the photocycle.
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Accepted/In Press date: 5 May 2021
Published date: 19 July 2021
Additional Information:
Funding Information:
We like to thank Heike Biehl for help with the purification of ChR2. The work was funded by Deutsche Forschungs-gemeinschaft/Sonderforschungsbereich 807 Transport and Communications across Membranes. The dynamic nuclear polarization experiments were enabled through DFG Equipment Grant GL 307/4-1 and the Cluster of Excellence Frankfurt: Macromolecular Complexes Frankfurt. Work at the Center for Biomolecular Magnetic Resonance is supported by the State of Hesse. Open access funding enabled and organized by Projekt DEAL.
Funding Information:
We like to thank Heike Biehl for help with the purification of ChR2. The work was funded by Deutsche Forschungs‐gemeinschaft/Sonderforschungsbereich 807 Transport and Communications across Membranes. The dynamic nuclear polarization experiments were enabled through DFG Equipment Grant GL 307/4‐1 and the Cluster of Excellence Frankfurt: Macromolecular Complexes Frankfurt. Work at the Center for Biomolecular Magnetic Resonance is supported by the State of Hesse. Open access funding enabled and organized by Projekt DEAL.
Publisher Copyright:
© 2021 The Authors. Angewandte Chemie International Edition published by Wiley-VCH GmbH
Keywords:
Photocycle, channelrhodopsin, dynamic nuclear polarization, membrane proteins, solid-state nmr spectroscopy
Identifiers
Local EPrints ID: 449543
URI: http://eprints.soton.ac.uk/id/eprint/449543
ISSN: 1433-7851
PURE UUID: e878e727-98ce-4e4c-9feb-add63b0cb4fc
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Date deposited: 07 Jun 2021 16:31
Last modified: 17 Mar 2024 02:47
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Contributors
Author:
Johanna Becker-Baldus
Author:
Alexander J Leeder
Author:
Christian Bamann
Author:
Clemens Glaubitz
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