A second photoactivatable state of the anion-conducting channelrhodopsin GtACR1 empowers persistent activity

Abstract Optogenetics is a method to regulate cells, tissues and organisms using light. It is applied to study neurons and to develop diagnostic and therapeutic tools for neuron-related diseases. The cation-conducting channelrhodopsin ChR2 triggers photoinduced depolarization of neuronal cells but g...

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Main Authors: Kristin Labudda, Mohamad Javad Norahan, Lisa-Marie Hübner, Philipp Althoff, Klaus Gerwert, Mathias Lübben, Till Rudack, Carsten Kötting
Format: Article
Language:English
Published: Nature Portfolio 2025-08-01
Series:Communications Biology
Online Access:https://doi.org/10.1038/s42003-025-08560-4
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author Kristin Labudda
Mohamad Javad Norahan
Lisa-Marie Hübner
Philipp Althoff
Klaus Gerwert
Mathias Lübben
Till Rudack
Carsten Kötting
author_facet Kristin Labudda
Mohamad Javad Norahan
Lisa-Marie Hübner
Philipp Althoff
Klaus Gerwert
Mathias Lübben
Till Rudack
Carsten Kötting
author_sort Kristin Labudda
collection DOAJ
description Abstract Optogenetics is a method to regulate cells, tissues and organisms using light. It is applied to study neurons and to develop diagnostic and therapeutic tools for neuron-related diseases. The cation-conducting channelrhodopsin ChR2 triggers photoinduced depolarization of neuronal cells but generates lower ion currents due to the syn-pathway of its branched photocycle. In contrast, the homologous anion-conducting ACR1 from Guillardia theta (GtACR1), exhibits high photocurrents. Here, we investigate the mechanistic cause for the observed high photocurrents in GtACR1 using FTIR spectroscopy. Unexpectedly, we discovered that the O intermediate of GtACR1 is photoactivable, allowing for fast and efficient channel reopening. Our vibrational spectra show a photocyclic reaction sequence after O excitation similar to the ground state photocycle but with slightly altered channel conformation and protonation states. Our results provide deeper insights into the gating mechanism of channelrhodopsins and pave the way to advance the development of optimized optogenetic tools in future.
format Article
id doaj-art-aa88609e5e594a75b5ccb0a69cc19e86
institution Kabale University
issn 2399-3642
language English
publishDate 2025-08-01
publisher Nature Portfolio
record_format Article
series Communications Biology
spelling doaj-art-aa88609e5e594a75b5ccb0a69cc19e862025-08-20T03:46:21ZengNature PortfolioCommunications Biology2399-36422025-08-018111210.1038/s42003-025-08560-4A second photoactivatable state of the anion-conducting channelrhodopsin GtACR1 empowers persistent activityKristin Labudda0Mohamad Javad Norahan1Lisa-Marie Hübner2Philipp Althoff3Klaus Gerwert4Mathias Lübben5Till Rudack6Carsten Kötting7Center for Protein Diagnostics (PRODI), Biospectroscopy, Ruhr University BochumCenter for Protein Diagnostics (PRODI), Biospectroscopy, Ruhr University BochumCenter for Protein Diagnostics (PRODI), Biospectroscopy, Ruhr University BochumCenter for Protein Diagnostics (PRODI), Biospectroscopy, Ruhr University BochumCenter for Protein Diagnostics (PRODI), Biospectroscopy, Ruhr University BochumCenter for Protein Diagnostics (PRODI), Biospectroscopy, Ruhr University BochumStructural Bioinformatics Group, Regensburg Center for Biochemistry, Regensburg Center for Ultrafast Nanoscopy, University of RegensburgCenter for Protein Diagnostics (PRODI), Biospectroscopy, Ruhr University BochumAbstract Optogenetics is a method to regulate cells, tissues and organisms using light. It is applied to study neurons and to develop diagnostic and therapeutic tools for neuron-related diseases. The cation-conducting channelrhodopsin ChR2 triggers photoinduced depolarization of neuronal cells but generates lower ion currents due to the syn-pathway of its branched photocycle. In contrast, the homologous anion-conducting ACR1 from Guillardia theta (GtACR1), exhibits high photocurrents. Here, we investigate the mechanistic cause for the observed high photocurrents in GtACR1 using FTIR spectroscopy. Unexpectedly, we discovered that the O intermediate of GtACR1 is photoactivable, allowing for fast and efficient channel reopening. Our vibrational spectra show a photocyclic reaction sequence after O excitation similar to the ground state photocycle but with slightly altered channel conformation and protonation states. Our results provide deeper insights into the gating mechanism of channelrhodopsins and pave the way to advance the development of optimized optogenetic tools in future.https://doi.org/10.1038/s42003-025-08560-4
spellingShingle Kristin Labudda
Mohamad Javad Norahan
Lisa-Marie Hübner
Philipp Althoff
Klaus Gerwert
Mathias Lübben
Till Rudack
Carsten Kötting
A second photoactivatable state of the anion-conducting channelrhodopsin GtACR1 empowers persistent activity
Communications Biology
title A second photoactivatable state of the anion-conducting channelrhodopsin GtACR1 empowers persistent activity
title_full A second photoactivatable state of the anion-conducting channelrhodopsin GtACR1 empowers persistent activity
title_fullStr A second photoactivatable state of the anion-conducting channelrhodopsin GtACR1 empowers persistent activity
title_full_unstemmed A second photoactivatable state of the anion-conducting channelrhodopsin GtACR1 empowers persistent activity
title_short A second photoactivatable state of the anion-conducting channelrhodopsin GtACR1 empowers persistent activity
title_sort second photoactivatable state of the anion conducting channelrhodopsin gtacr1 empowers persistent activity
url https://doi.org/10.1038/s42003-025-08560-4
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