Structural Mechanism of an Efficacy Photoswitch Targeting the β2-adrenergic Receptor

The field of photopharmacology develops light-responsive drugs that can modulate protein activity, enabling precise and dynamic investigations of their roles in health and disease. Adrenergic receptors are prominent targets for this approach because they are prototypical G protein-coupled receptors...

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Autores: Stipp, Robin, Bertrand, Quentin, Trabuco, Matilde, Duran-Corbera, Anna, Ignazzitto, Maria Tindara, Glover, Hannah, Stierli, Fabienne, Catena, Juanlo, Carrillo, Melissa, Hartmann, Sina, Seidel, Hans-Peter, Mulder, Matthias, Mason, Thomas, Kondo, Yasushi, Wranik, Maximillian, Appleby, Martin, Sager, Christoph, Sierra, Raymond, Gate, Gregory, Schleissner, Pamela, Cheng, Xinxin, Weinert, Tobias, Cheng, Robert, Mous, Sandra, Beale, John H., Kepa, Michal, Llebaria, Amadeu, Hennig, Michael, Rovira, Xavier, Standfuss, Joerg
Tipo de documento: artigo
Estado:Versão publicada
Data de publicação:2026
País:España
Recursos:Consejo Superior de Investigaciones Científicas (CSIC)
Repositório:DIGITAL.CSIC. Repositorio Institucional del CSIC
OAI Identifier:oai:dnet:digitalcsic_::e1f097cf1384d5a69e9a14d0ff922059
Acesso em linha:http://hdl.handle.net/10261/425703
Access Level:Acceso aberto
Palavra-chave:Time‐resolved serial femtosecond crystallography
G protein‐coupled receptors
Activity modulation
Azobenzene photoswitches
Photopharmacology
http://metadata.un.org/sdg/9
http://metadata.un.org/sdg/3
Ensure healthy lives and promote well-being for all at all ages
Build resilient infrastructure, promote inclusive and sustainable industrialization and foster innovation
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oai_identifier_str oai:dnet:digitalcsic_::e1f097cf1384d5a69e9a14d0ff922059
network_acronym_str ES
network_name_str España
repository_id_str
dc.title.none.fl_str_mv Structural Mechanism of an Efficacy Photoswitch Targeting the β2-adrenergic Receptor
title Structural Mechanism of an Efficacy Photoswitch Targeting the β2-adrenergic Receptor
spellingShingle Structural Mechanism of an Efficacy Photoswitch Targeting the β2-adrenergic Receptor
Stipp, Robin
Time‐resolved serial femtosecond crystallography
G protein‐coupled receptors
Activity modulation
Azobenzene photoswitches
Photopharmacology
http://metadata.un.org/sdg/9
http://metadata.un.org/sdg/3
Ensure healthy lives and promote well-being for all at all ages
Build resilient infrastructure, promote inclusive and sustainable industrialization and foster innovation
title_short Structural Mechanism of an Efficacy Photoswitch Targeting the β2-adrenergic Receptor
title_full Structural Mechanism of an Efficacy Photoswitch Targeting the β2-adrenergic Receptor
title_fullStr Structural Mechanism of an Efficacy Photoswitch Targeting the β2-adrenergic Receptor
title_full_unstemmed Structural Mechanism of an Efficacy Photoswitch Targeting the β2-adrenergic Receptor
title_sort Structural Mechanism of an Efficacy Photoswitch Targeting the β2-adrenergic Receptor
dc.creator.none.fl_str_mv Stipp, Robin
Bertrand, Quentin
Trabuco, Matilde
Duran-Corbera, Anna
Ignazzitto, Maria Tindara
Glover, Hannah
Stierli, Fabienne
Catena, Juanlo
Carrillo, Melissa
Hartmann, Sina
Seidel, Hans-Peter
Mulder, Matthias
Mason, Thomas
Kondo, Yasushi
Wranik, Maximillian
Appleby, Martin
Sager, Christoph
Sierra, Raymond
Gate, Gregory
Schleissner, Pamela
Cheng, Xinxin
Weinert, Tobias
Cheng, Robert
Mous, Sandra
Beale, John H.
Kepa, Michal
Llebaria, Amadeu
Hennig, Michael
Rovira, Xavier
Standfuss, Joerg
author Stipp, Robin
author_facet Stipp, Robin
Bertrand, Quentin
Trabuco, Matilde
Duran-Corbera, Anna
Ignazzitto, Maria Tindara
Glover, Hannah
Stierli, Fabienne
Catena, Juanlo
Carrillo, Melissa
Hartmann, Sina
Seidel, Hans-Peter
Mulder, Matthias
Mason, Thomas
Kondo, Yasushi
Wranik, Maximillian
Appleby, Martin
Sager, Christoph
Sierra, Raymond
Gate, Gregory
Schleissner, Pamela
Cheng, Xinxin
Weinert, Tobias
Cheng, Robert
Mous, Sandra
Beale, John H.
Kepa, Michal
Llebaria, Amadeu
Hennig, Michael
Rovira, Xavier
Standfuss, Joerg
author_role author
author2 Bertrand, Quentin
Trabuco, Matilde
Duran-Corbera, Anna
Ignazzitto, Maria Tindara
Glover, Hannah
Stierli, Fabienne
Catena, Juanlo
Carrillo, Melissa
Hartmann, Sina
Seidel, Hans-Peter
Mulder, Matthias
Mason, Thomas
Kondo, Yasushi
Wranik, Maximillian
Appleby, Martin
Sager, Christoph
Sierra, Raymond
Gate, Gregory
Schleissner, Pamela
Cheng, Xinxin
Weinert, Tobias
Cheng, Robert
Mous, Sandra
Beale, John H.
Kepa, Michal
Llebaria, Amadeu
Hennig, Michael
Rovira, Xavier
Standfuss, Joerg
author2_role author
author
author
author
author
author
author
author
author
author
author
author
author
author
author
author
author
author
author
author
author
author
author
author
author
author
author
author
author
dc.contributor.none.fl_str_mv Consejo Superior de Investigaciones Científicas [https://ror.org/02gfc7t72]
dc.subject.none.fl_str_mv Time‐resolved serial femtosecond crystallography
G protein‐coupled receptors
Activity modulation
Azobenzene photoswitches
Photopharmacology
http://metadata.un.org/sdg/9
http://metadata.un.org/sdg/3
Ensure healthy lives and promote well-being for all at all ages
Build resilient infrastructure, promote inclusive and sustainable industrialization and foster innovation
topic Time‐resolved serial femtosecond crystallography
G protein‐coupled receptors
Activity modulation
Azobenzene photoswitches
Photopharmacology
http://metadata.un.org/sdg/9
http://metadata.un.org/sdg/3
Ensure healthy lives and promote well-being for all at all ages
Build resilient infrastructure, promote inclusive and sustainable industrialization and foster innovation
description The field of photopharmacology develops light-responsive drugs that can modulate protein activity, enabling precise and dynamic investigations of their roles in health and disease. Adrenergic receptors are prominent targets for this approach because they are prototypical G protein-coupled receptors with high clinical relevance in bronchial and cardiovascular diseases. Here, we employed the azobenzene-based compound photoazolol-1 in combination with time-resolved serial crystallography at X-ray free-electron lasers to resolve the molecular mechanisms by which photoswitchable β-blockers modulate activity of the β2-adrenoceptor (β2AR). Time-resolved structures of the receptor bound to trans-photoazolol-1 (pre-photoconversion), a strained intermediate in the nanosecond range, and the fully photoisomerized cis-photoazolol-1 reveal how isomerization of the azobenzene moiety induces distinct conformational changes within the orthosteric ligand binding pocket. Within seconds, light-excited photoazolol-1 adopts a new binding pose, altering interactions with extracellular loop 2 and shifting the positions of transmembrane helices 5, 6, and 7. Functional assays of β2AR in cellular membranes show that photoazolol-1 acts as an efficacy photoswitch, changing from an inverse agonist to a neutral antagonist upon isomerization without leaving the binding pocket. In combination, these findings suggest a molecular mechanism for activity modulation via efficacy photoswitches and provide a framework for designing ligands that exploit light-driven transitions within the binding pocket to achieve spatiotemporal control of receptor function.
publishDate 2026
dc.date.none.fl_str_mv 2026
2026
2026
dc.type.none.fl_str_mv info:eu-repo/semantics/article
http://purl.org/coar/resource_type/c_6501
Publisher's version
info:eu-repo/semantics/publishedVersion
format article
status_str publishedVersion
dc.identifier.none.fl_str_mv http://hdl.handle.net/10261/425703
url http://hdl.handle.net/10261/425703
dc.language.none.fl_str_mv Inglés
language_invalid_str_mv Inglés
dc.relation.none.fl_str_mv Angewandte Chemie (International ed. in English)
https://doi.org/10.1002/anie.202517995

dc.rights.none.fl_str_mv info:eu-repo/semantics/openAccess
eu_rights_str_mv openAccess
dc.publisher.none.fl_str_mv Wiley-VCH
publisher.none.fl_str_mv Wiley-VCH
dc.source.none.fl_str_mv reponame:DIGITAL.CSIC. Repositorio Institucional del CSIC
instname:Consejo Superior de Investigaciones Científicas (CSIC)
instname_str Consejo Superior de Investigaciones Científicas (CSIC)
reponame_str DIGITAL.CSIC. Repositorio Institucional del CSIC
collection DIGITAL.CSIC. Repositorio Institucional del CSIC
repository.name.fl_str_mv
repository.mail.fl_str_mv
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spelling Structural Mechanism of an Efficacy Photoswitch Targeting the β2-adrenergic ReceptorStipp, RobinBertrand, QuentinTrabuco, MatildeDuran-Corbera, AnnaIgnazzitto, Maria TindaraGlover, HannahStierli, FabienneCatena, JuanloCarrillo, MelissaHartmann, SinaSeidel, Hans-PeterMulder, MatthiasMason, ThomasKondo, YasushiWranik, MaximillianAppleby, MartinSager, ChristophSierra, RaymondGate, GregorySchleissner, PamelaCheng, XinxinWeinert, TobiasCheng, RobertMous, SandraBeale, John H.Kepa, MichalLlebaria, AmadeuHennig, MichaelRovira, XavierStandfuss, JoergTime‐resolved serial femtosecond crystallographyG protein‐coupled receptorsActivity modulationAzobenzene photoswitchesPhotopharmacologyhttp://metadata.un.org/sdg/9http://metadata.un.org/sdg/3Ensure healthy lives and promote well-being for all at all agesBuild resilient infrastructure, promote inclusive and sustainable industrialization and foster innovationThe field of photopharmacology develops light-responsive drugs that can modulate protein activity, enabling precise and dynamic investigations of their roles in health and disease. Adrenergic receptors are prominent targets for this approach because they are prototypical G protein-coupled receptors with high clinical relevance in bronchial and cardiovascular diseases. Here, we employed the azobenzene-based compound photoazolol-1 in combination with time-resolved serial crystallography at X-ray free-electron lasers to resolve the molecular mechanisms by which photoswitchable β-blockers modulate activity of the β2-adrenoceptor (β2AR). Time-resolved structures of the receptor bound to trans-photoazolol-1 (pre-photoconversion), a strained intermediate in the nanosecond range, and the fully photoisomerized cis-photoazolol-1 reveal how isomerization of the azobenzene moiety induces distinct conformational changes within the orthosteric ligand binding pocket. Within seconds, light-excited photoazolol-1 adopts a new binding pose, altering interactions with extracellular loop 2 and shifting the positions of transmembrane helices 5, 6, and 7. Functional assays of β2AR in cellular membranes show that photoazolol-1 acts as an efficacy photoswitch, changing from an inverse agonist to a neutral antagonist upon isomerization without leaving the binding pocket. In combination, these findings suggest a molecular mechanism for activity modulation via efficacy photoswitches and provide a framework for designing ligands that exploit light-driven transitions within the binding pocket to achieve spatiotemporal control of receptor function.We are grateful for the support from the PSI Crystallization Facility and the Macromolecular Crystallography group during the growing and testing of crystals at the Swiss Light Source. The Biomolecular Structure and Mechanism Program of the Life Science Zürich Graduate School is acknowledged for its academic framework for our graduate students. Use of the Linac Coherent Light Source (LCLS), SLAC National Accelerator Laboratory, is supported by the U.S. Department of Energy, Office of Science, Office of Basic Energy Sciences under Contract No. DE-AC02-76SF00515. This work was supported by the National Institutes of Health grant S10 OD025079. Beamtime at the Cristallina station of the Swiss X-ray Free Electron laser (SwissFEL) was granted under proposal 20231114 in the first user run. The work was further funded by the Swiss National Science Foundation Project Grants 310030_197674 (to T.W.) and 310030_207462 (J.S.), the Swiss Innovation Agency Innosuisse Grant 42711.1 IP-LS (to M.H. and J.S.), the Swiss National Science Foundation Sinergia Grant CRSII5_213507 (J.S.), the Swiss Nanoscience Institute SNI #1904 (M.C.), the PSI Research Grant (to J.B. and J.S.), the Ministerio de Ciencia e Innovación and Agencia Estatal de Investigación and ERDF—A way of making Europe, PID2020-120499RB-I00 (AL and XR) and RYC2020-029485-I (XR), the Ministerio de Ciencia, Innovación y Universidades, PID2023-152950OB-I00 (AL) and PID2023-151614NB-I00 (XR), the Agència de Gestió d'Ajuts Universitaris i de Recerca (AGAUR)—Generalitat de Catalunya, 2021 SGR 00508 (AL) and the Spanish National Research Council, 20228AT014 (XR) Open access publishing facilitated by ETH-Bereich Forschungsanstalten, as part of the Wiley - ETH-Bereich Forschungsanstalten agreement via the Consortium Of Swiss Academic Libraries.Peer reviewedWiley-VCHConsejo Superior de Investigaciones Científicas [https://ror.org/02gfc7t72]202620262026info:eu-repo/semantics/articlehttp://purl.org/coar/resource_type/c_6501Publisher's versioninfo:eu-repo/semantics/publishedVersionhttp://hdl.handle.net/10261/425703reponame:DIGITAL.CSIC. Repositorio Institucional del CSICinstname:Consejo Superior de Investigaciones Científicas (CSIC)InglésAngewandte Chemie (International ed. in English)https://doi.org/10.1002/anie.202517995Síinfo:eu-repo/semantics/openAccessoai:dnet:digitalcsic_::e1f097cf1384d5a69e9a14d0ff9220592026-05-22T06:33:51Z
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