Project

Back to overview

Serial femtosecond crystallography of GPCR signaling systems

English title Serial femtosecond crystallography of GPCR signaling systems
Applicant Schertler Gebhard
Number 153145
Funding scheme Project funding
Research institution GPCR Signalling Biomolecular Research Paul Scherrer Institute
Institution of higher education Paul Scherrer Institute - PSI
Main discipline Molecular Biology
Start/End 01.06.2014 - 31.05.2017
Approved amount 834'000.00
Show all

All Disciplines (3)

Discipline
Molecular Biology
Biochemistry
Biophysics

Keywords (7)

free electron lasers; Subtype selectivity; serial femtosecond crystallography; G protein-coupled receptors; SwissFEL; GPCR signaling complex; Ligand specificity

Lay Summary (German)

Lead
Serielle femtosekunden Kristallographie von GPCR SignalsystemenProf Gebhard Schertler Bereichsleiter Biologie und Chemie, Paul Scherrer Institut (PSI)
Lay summary

G-Protein-gekoppelte Rezeptoren (GPCRs) sind mit mehr als 800 Mitgliedern im mensch­lichen Ge­nom kodiert. Aufgrund ihrer vielfältigen Funktionen in der zellulären Signal­weiterleitung sind sie Ziel etwa eines Drittel aller ver­füg­baren Medi­kamente. In ihrer zellulären Umgebung werden GPCRs von einer Vielzahl an kleinen Molekülen, den sogenannten GPCR Liganden, aktiviert. Jedoch binden sehr ähnliche GPCRs häufig dieselben Liganden, wodurch eine Reihe von physiologischen Signalen gleichzeitig ausgelöst werden kann. Ist der Ligand ein Medikament äussern sich die unerwünschten Signale dann in Nebenwirkungen. Das Medikament Risperidone gegen Schizophrenie z.B. bindet sowohl Adrenerge als auch Serotonin Rezeptoren. Ein besseres Verständnis dieser Prozesse kann daher Hinweise liefern wie Nebenwirkungen vermieden oder vermindert werden können.

In diesem Projekt werden wir Adrenerge und Serotonin Rezeptoren und ihre Signalkomplexe mittels der neuesten Technologie (serieller femtosekunden Kristallographie) durchleuchten um die Struktur und Bindung mit Liganden im molekularem Detail zu verstehen. Für dieses und andere Projekte aus Chemie, Physik und Biologie baut das Paul Scherrer Institut den Schweizer Freie-Elektronen Laser (SwissFEL). Ähnliche Grossforschungsanlagen gibt es zur Zeit nur in den USA und in Japan und ein weiteres erklärtes Ziel unserer Arbeit ist es mit unseren Experimenten von diesen existierenden Einrichtungen zu lernen um zukunftsträchtige Technologie besser in der Schweiz etablieren zu können. 

Direct link to Lay Summary Last update: 17.04.2014

Responsible applicant and co-applicants

Employees

Publications

Publication
Cryo-EM structure of the rhodopsin-Gαi-βγ complex reveals binding of the rhodopsin C-terminal tail to the gβ subunit
Tsai Ching-Ju, Marino Jacopo, Adaixo Ricardo, Pamula Filip, Muehle Jonas, Maeda Shoji, Flock Tilman, Taylor Nicholas MI, Mohammed Inayatulla, Matile Hugues, Dawson Roger JP, Deupi Xavier, Stahlberg Henning, Schertler Gebhard (2019), Cryo-EM structure of the rhodopsin-Gαi-βγ complex reveals binding of the rhodopsin C-terminal tail to the gβ subunit, in eLife, 8, 1-19.
Crystal structure of rhodopsin in complex with a mini-G o sheds light on the principles of G protein selectivity
Tsai Ching-Ju, Pamula Filip, Nehmé Rony, Mühle Jonas, Weinert Tobias, Flock Tilman, Nogly Przemyslaw, Edwards Patricia C., Carpenter Byron, Gruhl Thomas, Ma Pikyee, Deupi Xavier, Standfuss Jörg, Tate Christopher G., Schertler Gebhard F. X. (2018), Crystal structure of rhodopsin in complex with a mini-G o sheds light on the principles of G protein selectivity, in Science Advances, 4(9), eaat7052-eaat7052.
Retinal isomerization in bacteriorhodopsin captured by a femtosecond x-ray laser
Nogly Przemyslaw, Weinert Tobias, James Daniel, Carbajo Sergio, Ozerov Dmitry, Furrer Antonia, Gashi Dardan, Borin Veniamin, Skopintsev Petr, Jaeger Kathrin, Nass Karol, Båth Petra, Bosman Robert, Koglin Jason, Seaberg Matthew, Lane Thomas, Kekilli Demet, Brünle Steffen, Tanaka Tomoyuki, Wu Wenting, Milne Christopher, White Thomas, Barty Anton, Weierstall Uwe, et al. (2018), Retinal isomerization in bacteriorhodopsin captured by a femtosecond x-ray laser, in Science, eaat0094-eaat0094.
Ligand channel in pharmacologically stabilized rhodopsin
Mattle Daniel, Kuhn Bernd, Aebi Johannes, Bedoucha Marc, Kekilli Demet, Grozinger Nathalie, Alker Andre, Rudolph Markus G., Schmid Georg, Schertler Gebhard F. X., Hennig Michael, Standfuss Jörg, Dawson Roger J. P. (2018), Ligand channel in pharmacologically stabilized rhodopsin, in Proceedings of the National Academy of Sciences, 115(14), 3640-3645.
A three-dimensional movie of structural changes in bacteriorhodopsin
Nango Eriko, Royant Antoine, Kubo Minoru, Nakane Takanori, Wickstrand Cecilia, Kimura Tetsunari, Tanaka Tomoyuki, Tono Kensuke, Song Changyong, Tanaka Rie, Arima Toshi, Yamashita Ayumi, Kobayashi Jun, Hosaka Toshiaki, Mizohata Eiichi, Nogly Przemyslaw, Sugahara Michihiro, Nam Daewoong, Nomura Takashi, Shimamura Tatsuro, Im Dohyun, Fujiwara Takaaki, Yamanaka Yasuaki, Jeon Byeonghyun, et al. (2016), A three-dimensional movie of structural changes in bacteriorhodopsin, in Science, 354(6319), 1552-1557.
Structural role of the T94I rhodopsin mutation in congenital stationary night blindness
Singhal Ankita, Guo Ying, Matkovic Milos, Schertler Gebhard, Deupi Xavier, Yan Elsa CY, Standfuss Joerg (2016), Structural role of the T94I rhodopsin mutation in congenital stationary night blindness, in EMBO reports, 17(10), 1431-1440.
Lipidic cubic phase injector is a viable crystal delivery system for time-resolved serial crystallography
Nogly Przemyslaw, Panneels Valerie, Nelson Garrett, Gati Cornelius, Kimura Tetsunari, Milne Christopher, Milathianaki Despina, Kubo Minoru, Wu Wenting, Conrad Chelsie, Coe Jesse, Bean Richard, Zhao Yun, Båth Petra, Dods Robert, Harimoorthy Rajiv, Beyerlein Kenneth R., Rheinberger Jan, James Daniel, DePonte Daniel, Li Chufeng, Sala Leonardo, Williams Garth J., Hunter Mark S., et al. (2016), Lipidic cubic phase injector is a viable crystal delivery system for time-resolved serial crystallography, in Nature Communications, 7, 12314-12314.
Diverse activation pathways in class A GPCRs converge near the G-protein-coupling region
Venkatakrishnan A. J., Deupi Xavier, Lebon Guillaume, Heydenreich Franziska M., Flock Tilman, Miljus Tamara, Balaji Santhanam, Bouvier Michel, Veprintsev Dmitry B., Tate Christopher G., Schertler Gebhard F. X., Babu M. Madan (2016), Diverse activation pathways in class A GPCRs converge near the G-protein-coupling region, in Nature, 536(7617), 484-487.
Membrane protein structural biology using X-ray free electron lasers
Neutze Richard, Brändén Gisela, Schertler Gebhard FX (2015), Membrane protein structural biology using X-ray free electron lasers, in Current Opinion in Structural Biology, 33, 115-125.
Batch crystallization of rhodopsin for structural dynamics using an X-ray free-electron laser
Rheinberger Jan, Kick Leonhard M., Nelson Garrett, Deupi Xavier, Standfuss Jörg, Schertler Gebhard, Panneels Valérie, Wu Wenting, Nogly Przemyslaw, Gati Cornelius (2015), Batch crystallization of rhodopsin for structural dynamics using an X-ray free-electron laser, in Acta Crystallographica Section F Structural Biology Communications, 71(7), 856-860.
Time-resolved structural studies with serial crystallography: A new light on retinal proteins
Panneels Valérie, Wu Wenting, Tsai Ching-Ju, Nogly Przemek, Rheinberger Jan, Jaeger Kathrin, Cicchetti Gregor, Gati Cornelius, Kick Leonhard M., Sala Leonardo, Capitani Guido, Milne Chris, Padeste Celestino, Pedrini Bill, Li Xiao-Dan, Standfuss Jörg, Abela Rafael, Schertler Gebhard (2015), Time-resolved structural studies with serial crystallography: A new light on retinal proteins, in Structural Dynamics, 2(4), 041718-041718.
Lipidic cubic phase serial millisecond crystallography using synchrotron radiation
Nogly Przemyslaw, James Daniel, Wang Dingjie, White Thomas A., Zatsepin Nadia, Shilova Anastasya, Nelson Garrett, Liu Haiguang, Johansson Linda, Heymann Michael, Jaeger Kathrin, Metz Markus, Wickstrand Cecilia, Wu Wenting, Båth Petra, Berntsen Peter, Oberthuer Dominik, Panneels Valerie, Chapman Henry, Cherezov Vadim, Schertler Gebhard, Neutze Richard, Spence John, Moraes Isabel, et al. (2015), Lipidic cubic phase serial millisecond crystallography using synchrotron radiation, in IUCrJ, 2(2), 168-176.
Molecular mechanism of phosphorylation-dependent arrestin activation
Ostermaier Martin K, Schertler Gebhard FX, Standfuss Joerg (2014), Molecular mechanism of phosphorylation-dependent arrestin activation, in Current Opinion in Structural Biology, 29, 143-151.
Molecular mechanism of phosphorylation-dependent arrestin activation
Ostermaier Martin K, Schertler Gebhard FX, Standfuss Joerg (2014), Molecular mechanism of phosphorylation-dependent arrestin activation, in Current Opinion in Structural Biology, 143-151.
Serial millisecond crystallography for routine room- temperature structure determination at synchrotrons
Schertler Gebhard, Serial millisecond crystallography for routine room- temperature structure determination at synchrotrons, in NATURE COMMUNICATIONS , 8.

Collaboration

Group / person Country
Types of collaboration
Michael Hennig Switzerland (Europe)
- Industry/business/other use-inspired collaboration
Richard Neutze Sweden (Europe)
- in-depth/constructive exchanges on approaches, methods or results
- Publication
So Iwata Japan (Asia)
- in-depth/constructive exchanges on approaches, methods or results
- Publication
Linac Coherent Light Source (LCLS) United States of America (North America)
- in-depth/constructive exchanges on approaches, methods or results
- Research Infrastructure
Madan Babu Great Britain and Northern Ireland (Europe)
- in-depth/constructive exchanges on approaches, methods or results
- Publication

Scientific events

Active participation

Title Type of contribution Title of article or contribution Date Place Persons involved
"International Symposium on Biophysics of Rhodopsins: Celebrating Prof. Yoshinori Shichida" Talk given at a conference Keynote: Exploring GPCR and membrane protein dynamics with solution NMR and X-ray free-electron lasers 11.05.2017 Kyoto, Japan Schertler Gebhard;
52th Winter Seminar Talk given at a conference Exploring GPCR and membrane protein dynamics with solution NMR and X-ray free-electron lasers 04.01.2017 Kosters, Switzerland Schertler Gebhard;
16th International Conference on Retinal Proteins Poster Towards the study of the ultrafast rhodopsin photoactivation process using femtosecond serial crystallograpy 02.10.2016 Potsdam, Germany Gruhl Thomas; Mühle Jonas;
16th International Conference on Retinal Proteins Talk given at a conference Session chair and intro: Non-visual rhodopsins and synthetic photoreceptors 02.10.2016 Potsdam, Germany Schertler Gebhard;
14th Annual Discovery on Target Talk given at a conference Keynote: NMR AND ALLOSTERIC SIGNAL TRANSDUCTION NETWORKS IN THE β1- ADRENERGIC RECEPTOR 19.09.2016 Boston, United States of America Schertler Gebhard;
2nd European Meeting on Phototransduction Talk given at a conference The structure of visual pigments: implications for rare GPCR diseases 04.09.2016 Monte Veritas, Switzerland Schertler Gebhard;
FASEB 2016 “Molecular Biophysics of Membranes” Talk given at a conference Structural Biology of GPCRs: The impact of NMR and x-ray Free Electron Lasers 10.07.2016 Snowmass, CO, United States of America Schertler Gebhard;
D-BIOL Symposium 2016 Poster Study of the ultrafast rhodopsin photoactivation process using femtosecond serial crystallograpy 13.06.2016 Davos, Switzerland Mühle Jonas;
3rd NovAliX Conference Talk given at a conference Exploring GPCR and Membrane Protein Dynamics with Solution NMR and X-ray Free-Electron Lasers 07.06.2016 Strasbourg, France Schertler Gebhard;
8th International Adhesion GPCR Workshop Talk given at a conference Plenary Lecture: Exploring GPCR and membrane protein dynamics with solution NMR and X-ray free-electron lasers 02.06.2016 Leipzig, Germany Schertler Gebhard;
GLISTEN Meeting 2016 Talk given at a conference The application of Free Electron Lasers to biology: a new age of time resolved crystallography 06.04.2016 Erlangen, Germany Piscitelli Chayne; Schertler Gebhard;
2nd SACLA Workshop on Structural Biology with FELs Talk given at a conference Update on some recent time-resolved experiments performedat SACLA & LCLS 02.02.2016 RIKEN SPring-8 Center , Japan Gruhl Thomas; Wu Wenting;
Gordon conference: Ligand Recognition & Molecular Gating Talk given at a conference "The Application of Free Electron Lasers to Biology: A New Age of Time-Resolved Crystallography" 31.01.2016 Lucca, Italy Schertler Gebhard;
2016 Senri Life Science International Symposium "Frontiers in Structural Biology --- X-ray Free Electron Laser and Drug Discovery" Talk given at a conference The application of Free Electron Lasers to biology: a new age of time resolved crystallography 22.01.2016 Osaka, Japan Schertler Gebhard;
Pacifichem 2015 Talk given at a conference From the structure of vertebrate and invertebrate rhodopsins to new applications in optogenetics 15.12.2015 Honolulu, Hawaii, United States of America Schertler Gebhard;
FIMPSB 2015 Frontiers in Integral Membrane Protein Structural Biology Talk given at a conference The impact of Free Electron Lasers on membrane protein structural biology 02.10.2015 Oxford, Great Britain and Northern Ireland Schertler Gebhard;


Self-organised

Title Date Place
Hybrid Methods Workshop 21.03.2016 Paul Scherrer Institut, Switzerland

Knowledge transfer events

Active participation

Title Type of contribution Date Place Persons involved
UCB Structural Biology Symposium Talk 19.11.2015 Windsor, Great Britain and Northern Ireland Schertler Gebhard;
Silicon Valley Meets Switzerland Talk 12.03.2015 Brugg, Switzerland Schertler Gebhard;


Communication with the public

Communication Title Media Place Year
Talks/events/exhibitions Mit Röntgenstrahlen die Geheimnisse des Lebens entschlüsseln German-speaking Switzerland 2015

Use-inspired outputs


Start-ups

Name Year
InterAx 2016
leadXpro 2015

Associated projects

Number Title Start Funding scheme
132815 Linking G protein-coupled receptor structure to signaling output 01.12.2010 Project funding
132815 Linking G protein-coupled receptor structure to signaling output 01.12.2010 Project funding
192760 Structure based engineering of photo-catalysis using Free Electron Lasers 01.06.2020 Project funding
141898 NMR studies of GPCRs: Structure, dynamics and interactions with ligands and signaling proteins 01.12.2012 Sinergia
173335 Analyzing protein dynamics in receptor signaling relevant to pharmacology 01.06.2017 Project funding
170802 Direct electron detector for cryo-EM single particle analysis, electron tomography and protein nanocrystallography 01.11.2017 R'EQUIP
177125 Tunable nanosecond laser for time-resolved serial crystallography at SLS and SwissFEL 01.01.2018 R'EQUIP

Abstract

G protein-coupled receptors (GPCRs) comprise 2-3% of human proteins and are key for cellular communication in higher organisms. These membrane receptors operate by ligand-dependent coupling to several intracellular effector proteins, including G proteins, kinases and arrestins. GPCR signaling is often modulated by a series of closely related receptors that vary in their relative affinity to natural ligands and pharmaceutical drugs. Cross-reactivity between such related receptors is a common cause of side effects, which reduces the benefits of many medical drugs.The goal of this project is to gain a better understanding of the molecular basis of ligand selectivity in serotonin and adrenergic receptors, both members of the amine family of GPCRs. My group has a long experience in the structure determination of GPCRs, in particular rhodopsin and the ß1 and ß2 adrenergic receptors. Here we propose to target all nine adrenergic receptor subtypes to obtain a set as complete as possible of human GPCR subtypes. The structure of the related amine receptor 5-HTc will provide further templates to study GPCR ligand selectivity. For instance, the antipsychotic drug risperidone binds both adrenergic as well as serotonin receptors with high affinity. Thus, crystal structures of several amine receptor subtypes bound to this inverse agonist may provide important clues on how to prevent drug cross-reactivity.In addition, a deeper understanding of the molecular mechanisms of GPCR activation requires structural knowledge of the complexes between the receptors and their effector molecules. This proposal also aims to solve the structure of the complex between the GPCR rhodopsin, responsible for vision in dim-light conditions, and the Gt and Gi proteins.The recent development of serial femtosecond nanocrystallography using free electron lasers has opened the door for high-throughput structure determination of closely related proteins or protein-ligand complexes. The use of lipidic cubic phases for crystal growth and delivery significantly reduces the amount of protein needed for such experiments. Importantly, data collection using highly brilliant femtosecond laser pulses allows to outrun radiation damage and determine membrane protein structures at previously unprecedented resolution. GPCRs bound to many different pharmacologically important ligands, and in complex with G proteins, are the ideal targets to establish serial femtosecond nanocrystallography for membrane proteins in Switzerland. This project will therefore allow us to be fully prepared for a new user community when the X-ray laser SwissFEL becomes operational at the Paul Scherrer Institute.
-