Crystal structure of a natural light-gated anion channelrhodopsin.

Hai Li; Chia-Ying Huang; Elena Govorunova; Chris Schafer; Oleg Sineshchekov; Meitian Wang; Lei Zheng; John Spudich

doi:10.7554/elife.41741

Crystal structure of a natural light-gated anion channelrhodopsin.

Hai Li
, Chia-Ying Huang
, Elena Govorunova
, Chris Schafer
, Oleg Sineshchekov
, Meitian Wang
, Lei Zheng^*
, John Spudich^*

^*Corresponding author for this work

Research output: Contribution to Journal › Article › Academic › peer-review

Abstract

The anion channelrhodopsin GtACR1 from the alga Guillardia theta is a potent neuron-inhibiting optogenetics tool. Presented here, its X-ray structure at 2.9 Å reveals a tunnel traversing the protein from its extracellular surface to a large cytoplasmic cavity. The tunnel is lined primarily by small polar and aliphatic residues essential for anion conductance. A disulfide-immobilized extracellular cap facilitates channel closing and the ion path is blocked mid-membrane by its photoactive retinylidene chromophore and further by a cytoplasmic side constriction. The structure also reveals a novel photoactive site configuration that maintains the retinylidene Schiff base protonated when the channel is open. These findings suggest a new channelrhodopsin mechanism, in which the Schiff base not only controls gating, but also serves as a direct mediator for anion flux.

Original language	English
Article number	e41741
Pages (from-to)	1-21
Journal	eLife
Volume	8
DOIs	https://doi.org/10.7554/elife.41741
Publication status	Published - 7 Jan 2019

Funding

This work was supported by National Institutes of Health Grants R01GM027750 and U01MH109146, the Hermann Eye Fund, and Endowed Chair AU-0009 from the Robert A Welch Foundation to JLS, and American Heart Association Grant 18TPA34230046 to LZ. C-YH was partially supported by the European Union’s Horizon 2020 research and innovation programme under the Marie-Skłodowska-Curie grant agreement No. 701647. This work was supported by National Institutes of Health Grants R01GM027750 and U01MH109146, the Hermann Eye Fund, and Endowed Chair AU-0009 from the Robert A Welch Foundation to JLS, and American Heart Association Grant 18TPA34230046 to LZ. C-YH was partially supported by the European Union?s Horizon 2020 research and innovation programme under the Marie-Sk?odowska-Curie grant agreement No. 701647. National Institutes of Health R01GM027750 John L Spudich National Institutes of Health U01MH109146 John L Spudich Welch Foundation AU-0009 John L Spudich American Heart Association 18TPA34230046 Lei Zheng Hermann Eye Fund John L Spudich H2020 Marie Sk?odowska-Curie Actions 701647 Chia-Ying Huang The funders had no role in study design, data collection and interpretation, or the decision to submit the work for publication.

Funders	Funder number
National Institutes of Health R01GM027750 John L Spudich National Institutes of Health U01MH109146 John L Spudich Welch Foundation AU-0009 John L Spudich American Heart Association 18TPA34230046 Lei Zheng Hermann Eye Fund John L Spudich H2020 Marie Sk?odowska
National Institutes of Health	R01GM027750
National Institute of Mental Health	U01MH109146
Welch Foundation
American Heart Association	18TPA34230046
Horizon 2020 Framework Programme	701647
Hermann Eye Fund	AU-0009

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title = "Crystal structure of a natural light-gated anion channelrhodopsin.",

abstract = "The anion channelrhodopsin GtACR1 from the alga Guillardia theta is a potent neuron-inhibiting optogenetics tool. Presented here, its X-ray structure at 2.9 {\AA} reveals a tunnel traversing the protein from its extracellular surface to a large cytoplasmic cavity. The tunnel is lined primarily by small polar and aliphatic residues essential for anion conductance. A disulfide-immobilized extracellular cap facilitates channel closing and the ion path is blocked mid-membrane by its photoactive retinylidene chromophore and further by a cytoplasmic side constriction. The structure also reveals a novel photoactive site configuration that maintains the retinylidene Schiff base protonated when the channel is open. These findings suggest a new channelrhodopsin mechanism, in which the Schiff base not only controls gating, but also serves as a direct mediator for anion flux.",

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AU - Li, Hai

AU - Huang, Chia-Ying

AU - Govorunova, Elena

AU - Schafer, Chris

AU - Sineshchekov, Oleg

AU - Wang, Meitian

AU - Zheng, Lei

AU - Spudich, John

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AB - The anion channelrhodopsin GtACR1 from the alga Guillardia theta is a potent neuron-inhibiting optogenetics tool. Presented here, its X-ray structure at 2.9 Å reveals a tunnel traversing the protein from its extracellular surface to a large cytoplasmic cavity. The tunnel is lined primarily by small polar and aliphatic residues essential for anion conductance. A disulfide-immobilized extracellular cap facilitates channel closing and the ion path is blocked mid-membrane by its photoactive retinylidene chromophore and further by a cytoplasmic side constriction. The structure also reveals a novel photoactive site configuration that maintains the retinylidene Schiff base protonated when the channel is open. These findings suggest a new channelrhodopsin mechanism, in which the Schiff base not only controls gating, but also serves as a direct mediator for anion flux.

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Crystal structure of a natural light-gated anion channelrhodopsin.

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