Structural insights into light-gating of potassium-selective channelrhodopsin
Abstract Structural information on channelrhodopsins’ mechanism of light-gated ion conductance is scarce, limiting its engineering as optogenetic tools. Here, we use single-particle cryo-electron microscopy of peptidisc-incorporated protein samples to determine the structures of the slow-cycling mut...
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Nature Portfolio
2025-02-01
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| Series: | Nature Communications |
| Online Access: | https://doi.org/10.1038/s41467-025-56491-9 |
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| author | Takefumi Morizumi Kyumhyuk Kim Hai Li Probal Nag Tal Dogon Oleg A. Sineshchekov Yumei Wang Leonid S. Brown Songhwan Hwang Han Sun Ana-Nicoleta Bondar Igor Schapiro Elena G. Govorunova John L. Spudich Oliver P. Ernst |
| author_facet | Takefumi Morizumi Kyumhyuk Kim Hai Li Probal Nag Tal Dogon Oleg A. Sineshchekov Yumei Wang Leonid S. Brown Songhwan Hwang Han Sun Ana-Nicoleta Bondar Igor Schapiro Elena G. Govorunova John L. Spudich Oliver P. Ernst |
| author_sort | Takefumi Morizumi |
| collection | DOAJ |
| description | Abstract Structural information on channelrhodopsins’ mechanism of light-gated ion conductance is scarce, limiting its engineering as optogenetic tools. Here, we use single-particle cryo-electron microscopy of peptidisc-incorporated protein samples to determine the structures of the slow-cycling mutant C110A of kalium channelrhodopsin 1 from Hyphochytrium catenoides (HcKCR1) in the dark and upon laser flash excitation. Upon photoisomerization of the retinal chromophore, the retinylidene Schiff base NH-bond reorients from the extracellular to the cytoplasmic side. This switch triggers a series of side chain reorientations and merges intramolecular cavities into a transmembrane K+ conduction pathway. Molecular dynamics simulations confirm K+ flux through the illuminated state but not through the resting state. The overall displacement between the closed and the open structure is small, involving mainly side chain rearrangements. Asp105 and Asp116 play a key role in K+ conductance. Structure-guided mutagenesis and patch-clamp analysis reveal the roles of the pathway-forming residues in channel gating and selectivity. |
| format | Article |
| id | doaj-art-152a65bfbbe54ce695fd0661c128d0f4 |
| institution | Kabale University |
| issn | 2041-1723 |
| language | English |
| publishDate | 2025-02-01 |
| publisher | Nature Portfolio |
| record_format | Article |
| series | Nature Communications |
| spelling | doaj-art-152a65bfbbe54ce695fd0661c128d0f42025-02-09T12:45:31ZengNature PortfolioNature Communications2041-17232025-02-0116111510.1038/s41467-025-56491-9Structural insights into light-gating of potassium-selective channelrhodopsinTakefumi Morizumi0Kyumhyuk Kim1Hai Li2Probal Nag3Tal Dogon4Oleg A. Sineshchekov5Yumei Wang6Leonid S. Brown7Songhwan Hwang8Han Sun9Ana-Nicoleta Bondar10Igor Schapiro11Elena G. Govorunova12John L. Spudich13Oliver P. Ernst14Department of Biochemistry, University of TorontoDepartment of Biochemistry, University of TorontoDepartment of Biochemistry & Molecular Biology, Center for Membrane Biology, The University of Texas Health Science Center at Houston McGovern Medical SchoolInstitute of Chemistry, The Hebrew University of JerusalemInstitute of Chemistry, The Hebrew University of JerusalemDepartment of Biochemistry & Molecular Biology, Center for Membrane Biology, The University of Texas Health Science Center at Houston McGovern Medical SchoolDepartment of Biochemistry & Molecular Biology, Center for Membrane Biology, The University of Texas Health Science Center at Houston McGovern Medical SchoolDepartment of Physics and Biophysics Interdepartmental Group, University of GuelphLeibniz-Forschungsinstitut für Molekulare Pharmakologie (FMP)Leibniz-Forschungsinstitut für Molekulare Pharmakologie (FMP)Faculty of Physics, University of BucharestInstitute of Chemistry, The Hebrew University of JerusalemDepartment of Biochemistry & Molecular Biology, Center for Membrane Biology, The University of Texas Health Science Center at Houston McGovern Medical SchoolDepartment of Biochemistry & Molecular Biology, Center for Membrane Biology, The University of Texas Health Science Center at Houston McGovern Medical SchoolDepartment of Biochemistry, University of TorontoAbstract Structural information on channelrhodopsins’ mechanism of light-gated ion conductance is scarce, limiting its engineering as optogenetic tools. Here, we use single-particle cryo-electron microscopy of peptidisc-incorporated protein samples to determine the structures of the slow-cycling mutant C110A of kalium channelrhodopsin 1 from Hyphochytrium catenoides (HcKCR1) in the dark and upon laser flash excitation. Upon photoisomerization of the retinal chromophore, the retinylidene Schiff base NH-bond reorients from the extracellular to the cytoplasmic side. This switch triggers a series of side chain reorientations and merges intramolecular cavities into a transmembrane K+ conduction pathway. Molecular dynamics simulations confirm K+ flux through the illuminated state but not through the resting state. The overall displacement between the closed and the open structure is small, involving mainly side chain rearrangements. Asp105 and Asp116 play a key role in K+ conductance. Structure-guided mutagenesis and patch-clamp analysis reveal the roles of the pathway-forming residues in channel gating and selectivity.https://doi.org/10.1038/s41467-025-56491-9 |
| spellingShingle | Takefumi Morizumi Kyumhyuk Kim Hai Li Probal Nag Tal Dogon Oleg A. Sineshchekov Yumei Wang Leonid S. Brown Songhwan Hwang Han Sun Ana-Nicoleta Bondar Igor Schapiro Elena G. Govorunova John L. Spudich Oliver P. Ernst Structural insights into light-gating of potassium-selective channelrhodopsin Nature Communications |
| title | Structural insights into light-gating of potassium-selective channelrhodopsin |
| title_full | Structural insights into light-gating of potassium-selective channelrhodopsin |
| title_fullStr | Structural insights into light-gating of potassium-selective channelrhodopsin |
| title_full_unstemmed | Structural insights into light-gating of potassium-selective channelrhodopsin |
| title_short | Structural insights into light-gating of potassium-selective channelrhodopsin |
| title_sort | structural insights into light gating of potassium selective channelrhodopsin |
| url | https://doi.org/10.1038/s41467-025-56491-9 |
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