Structural basis for the prolonged photocycle of sensory rhodopsin II revealed by serial synchrotron crystallography

Abstract Microbial rhodopsins form a diverse family of light-sensitive seven-transmembrane helix retinal proteins that function as active proton or ion pumps, passive light-gated ion channels, and photosensors. To understand how light-sensing in archaea is initiated by sensory rhodopsins, we perform...

Full description

Saved in:
Bibliographic Details
Main Authors: Robert Bosman, Giorgia Ortolani, Swagatha Ghosh, Daniel James, Per Norder, Greger Hammarin, Tinna Björg Úlfarsdóttir, Lucija Ostojić, Tobias Weinert, Florian Dworkowski, Takashi Tomizaki, Jörg Standfuss, Gisela Brändén, Richard Neutze
Format: Article
Language:English
Published: Nature Portfolio 2025-04-01
Series:Nature Communications
Online Access:https://doi.org/10.1038/s41467-025-58263-x
Tags: Add Tag
No Tags, Be the first to tag this record!
_version_ 1850145549029736448
author Robert Bosman
Giorgia Ortolani
Swagatha Ghosh
Daniel James
Per Norder
Greger Hammarin
Tinna Björg Úlfarsdóttir
Lucija Ostojić
Tobias Weinert
Florian Dworkowski
Takashi Tomizaki
Jörg Standfuss
Gisela Brändén
Richard Neutze
author_facet Robert Bosman
Giorgia Ortolani
Swagatha Ghosh
Daniel James
Per Norder
Greger Hammarin
Tinna Björg Úlfarsdóttir
Lucija Ostojić
Tobias Weinert
Florian Dworkowski
Takashi Tomizaki
Jörg Standfuss
Gisela Brändén
Richard Neutze
author_sort Robert Bosman
collection DOAJ
description Abstract Microbial rhodopsins form a diverse family of light-sensitive seven-transmembrane helix retinal proteins that function as active proton or ion pumps, passive light-gated ion channels, and photosensors. To understand how light-sensing in archaea is initiated by sensory rhodopsins, we perform serial synchrotron X-ray crystallography (SSX) studies of light induced conformational changes in sensory rhodopsin II (NpSRII) from the archaea Natronomonas pharaonis, both collecting time-resolved SSX data and collecting SSX data during continuous illumination. Comparing light-induced electron density changes in NpSRII with those reported for bacteriorhodopsin (bR) reveals several common light-induced structural perturbations. Unlike bR, however, helix G of NpSRII does not unwind near the conserved lysine residue to which retinal is covalently bound and therefore transient water molecule binding sites do not arise immediately to the cytoplasmic side of retinal. These structural differences prolong the duration of the NpSRII photocycle relative to bR, allowing time for the light-initiated sensory signal to be amplified.
format Article
id doaj-art-b8d39a98c721407ca768780e78ecb9b2
institution OA Journals
issn 2041-1723
language English
publishDate 2025-04-01
publisher Nature Portfolio
record_format Article
series Nature Communications
spelling doaj-art-b8d39a98c721407ca768780e78ecb9b22025-08-20T02:28:04ZengNature PortfolioNature Communications2041-17232025-04-0116111210.1038/s41467-025-58263-xStructural basis for the prolonged photocycle of sensory rhodopsin II revealed by serial synchrotron crystallographyRobert Bosman0Giorgia Ortolani1Swagatha Ghosh2Daniel James3Per Norder4Greger Hammarin5Tinna Björg Úlfarsdóttir6Lucija Ostojić7Tobias Weinert8Florian Dworkowski9Takashi Tomizaki10Jörg Standfuss11Gisela Brändén12Richard Neutze13Department of Chemistry and Molecular Biology, University of GothenburgDepartment of Chemistry and Molecular Biology, University of GothenburgDepartment of Chemistry and Molecular Biology, University of GothenburgLaboratory of Biomolecular Research, Center for Life Sciences, Paul Scherrer Institut, Forschungsstrasse 111Department of Chemistry and Molecular Biology, University of GothenburgDepartment of Chemistry and Molecular Biology, University of GothenburgDepartment of Chemistry and Molecular Biology, University of GothenburgDepartment of Chemistry and Molecular Biology, University of GothenburgLaboratory of Biomolecular Research, Center for Life Sciences, Paul Scherrer Institut, Forschungsstrasse 111Laboratory of Femtochemistry, Center for Photon Science, Paul Scherrer Institut, Forschungsstrasse 111Laboratory of Macromolecules and Bioimaging, Center for Photon Science, Paul Scherrer Institut, Forschungsstrasse 111Laboratory of Biomolecular Research, Center for Life Sciences, Paul Scherrer Institut, Forschungsstrasse 111Department of Chemistry and Molecular Biology, University of GothenburgDepartment of Chemistry and Molecular Biology, University of GothenburgAbstract Microbial rhodopsins form a diverse family of light-sensitive seven-transmembrane helix retinal proteins that function as active proton or ion pumps, passive light-gated ion channels, and photosensors. To understand how light-sensing in archaea is initiated by sensory rhodopsins, we perform serial synchrotron X-ray crystallography (SSX) studies of light induced conformational changes in sensory rhodopsin II (NpSRII) from the archaea Natronomonas pharaonis, both collecting time-resolved SSX data and collecting SSX data during continuous illumination. Comparing light-induced electron density changes in NpSRII with those reported for bacteriorhodopsin (bR) reveals several common light-induced structural perturbations. Unlike bR, however, helix G of NpSRII does not unwind near the conserved lysine residue to which retinal is covalently bound and therefore transient water molecule binding sites do not arise immediately to the cytoplasmic side of retinal. These structural differences prolong the duration of the NpSRII photocycle relative to bR, allowing time for the light-initiated sensory signal to be amplified.https://doi.org/10.1038/s41467-025-58263-x
spellingShingle Robert Bosman
Giorgia Ortolani
Swagatha Ghosh
Daniel James
Per Norder
Greger Hammarin
Tinna Björg Úlfarsdóttir
Lucija Ostojić
Tobias Weinert
Florian Dworkowski
Takashi Tomizaki
Jörg Standfuss
Gisela Brändén
Richard Neutze
Structural basis for the prolonged photocycle of sensory rhodopsin II revealed by serial synchrotron crystallography
Nature Communications
title Structural basis for the prolonged photocycle of sensory rhodopsin II revealed by serial synchrotron crystallography
title_full Structural basis for the prolonged photocycle of sensory rhodopsin II revealed by serial synchrotron crystallography
title_fullStr Structural basis for the prolonged photocycle of sensory rhodopsin II revealed by serial synchrotron crystallography
title_full_unstemmed Structural basis for the prolonged photocycle of sensory rhodopsin II revealed by serial synchrotron crystallography
title_short Structural basis for the prolonged photocycle of sensory rhodopsin II revealed by serial synchrotron crystallography
title_sort structural basis for the prolonged photocycle of sensory rhodopsin ii revealed by serial synchrotron crystallography
url https://doi.org/10.1038/s41467-025-58263-x
work_keys_str_mv AT robertbosman structuralbasisfortheprolongedphotocycleofsensoryrhodopsiniirevealedbyserialsynchrotroncrystallography
AT giorgiaortolani structuralbasisfortheprolongedphotocycleofsensoryrhodopsiniirevealedbyserialsynchrotroncrystallography
AT swagathaghosh structuralbasisfortheprolongedphotocycleofsensoryrhodopsiniirevealedbyserialsynchrotroncrystallography
AT danieljames structuralbasisfortheprolongedphotocycleofsensoryrhodopsiniirevealedbyserialsynchrotroncrystallography
AT pernorder structuralbasisfortheprolongedphotocycleofsensoryrhodopsiniirevealedbyserialsynchrotroncrystallography
AT gregerhammarin structuralbasisfortheprolongedphotocycleofsensoryrhodopsiniirevealedbyserialsynchrotroncrystallography
AT tinnabjorgulfarsdottir structuralbasisfortheprolongedphotocycleofsensoryrhodopsiniirevealedbyserialsynchrotroncrystallography
AT lucijaostojic structuralbasisfortheprolongedphotocycleofsensoryrhodopsiniirevealedbyserialsynchrotroncrystallography
AT tobiasweinert structuralbasisfortheprolongedphotocycleofsensoryrhodopsiniirevealedbyserialsynchrotroncrystallography
AT floriandworkowski structuralbasisfortheprolongedphotocycleofsensoryrhodopsiniirevealedbyserialsynchrotroncrystallography
AT takashitomizaki structuralbasisfortheprolongedphotocycleofsensoryrhodopsiniirevealedbyserialsynchrotroncrystallography
AT jorgstandfuss structuralbasisfortheprolongedphotocycleofsensoryrhodopsiniirevealedbyserialsynchrotroncrystallography
AT giselabranden structuralbasisfortheprolongedphotocycleofsensoryrhodopsiniirevealedbyserialsynchrotroncrystallography
AT richardneutze structuralbasisfortheprolongedphotocycleofsensoryrhodopsiniirevealedbyserialsynchrotroncrystallography