Dr. Jörg Standfuss

Photo of Jörg Standfuss

Group Leader

Paul Scherrer Institute
Forschungsstrasse 111
5232 Villigen PSI
Suisse

Serial crystallography using synchrotron radiation and free electron lasers

One of the major promises of X-ray free-electron laser (XFEL) technology is to advance structural biology from the determination of molecular snapshots to molecular movies. Together structural and dynamic information provides unique insights into the function of proteins as principal building blocks of our biology.



The key advantage of XFEL-based over conventional diffraction strategies stems from the characteristics of the XFEL pulses, which contain ~1012 X-ray photons but are only tens of femtoseconds long. This fluence is sufficient to produce high-resolution diffraction patterns from very small crystals, while at the same time outrunning most radiation damage processes. The ultrafast XFEL pulses further open up the possibility for time-resolved pump probe experiments on femtosecond to millisecond timescales. Together these advantages allow studying protein structural dynamics at unprecedented spatial and temporal resolution; essentially opening a new frontier in structural biology.

My group focuses on the implementation of time-resolved serial crystallography at the Swiss Light Source (SLS) and the Swiss Free Electron Laser (SwissFEL). Naturally light-sensitive retinal-binding proteins like bacteriorhodopsin are paving the way for time-resolved studies using XFELs. In the near future, we will further see the molecular details of how protons, chloride and sodium ions are pumped across biological membranes by light. My research further aims to answer the question of how protein interactions guide the high quantum efficiency and stereo selectivity of the ultrafast retinal isomerization. Time-resolved crystallography will help us understand fundamental biological processes including the high photo efficiency of the visual sense and how retinal proteins might be improved to better manipulate neural cells in the field of optogenetics.

Laser light allows unmatched precision which makes it an ideal trigger for time-resolved measurements. Learning from biology, chemists have developed a large repertoire of synthetic photoswitches with highly tunable properties. Like their natural counterpart retinal, these chromophores can be inserted into proteins to put them under optical control. In photopharmacological applications reversibly binding ligands are envisioned to precisely control pharmaceutical targets such as ion channels, GPCRs or microtubules. Compared to optogenetics the approach has a wider range of applications including medical intervention in humans since the method relies on the chemical manipulation of native proteins and is not dependent on genetic manipulation. Harnessing the chemistry of such synthetic photoswitches to study proteins that cannot be natively activated by light will dramatically increase the number of biological systems whose structural dynamics can be studied at modern XFEL sources. Dynamic information on how ligands influence a particular protein conformation will provide a crucial new dimension to molecular pharmacology.

Jul 2019, Muotatal and Hölloch Exploration

Exploration Muotatal and Hölloch

Jul 2019,  Time-resolved crystallography now possible at the SLS
https://www.psi.ch/en/media/our-research/molecular-energy-machine-as-a-movie-star

Oct 2018, Climbing at high wire park Pilatus

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Nov 2017, Przemek Nogly started group at the ETH Zurich on a SNSF Ambizione grant. Congratulations well deserved!!

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Sep 2017, Demet Kekilli and Steffen Brünle joined the PSI-FELLOW-II programme co-funded by Horizon2020 of the European Commission

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Jan 2017, Our research featured by the Swiss National Science Foundation

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Aug 2016, PSI Press Release "Catching Proteins in the Act"
https://www.psi.ch/media/catching-proteins-in-the-act

Jul 2016, Besserstein Hike

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Jaeger,Kathrin
Ph.D. Student

Mattle,Daniel Dr.
Postdoc

Nogly,Przemek Dr.
Postdoc

Peterhans,Christian
Ph.D. Student

Singhal,Ankita
Ph.D. Student



Publications since 2010

  • Haider RS, Wilhelm F, Rizk A, Mutt E, Deupi X, Peterhans C, et al.
    Arrestin-1 engineering facilitates complex stabilization with native rhodopsin
    Scientific Reports. 2019; 9(1): 439 (13 pp.). https://doi.org/10.1038/s41598-018-36881-4
    DORA PSI
  • Jaeger K, Bruenle S, Weinert T, Guba W, Muehle J, Miyazaki T, et al.
    Structural basis for allosteric ligand recognition in the human CC chemokine receptor 7
    Cell. 2019; 178(5): 1222-1230. https://doi.org/10.1016/j.cell.2019.07.028
    DORA PSI
  • James D, Weinert T, Skopintsev P, Furrer A, Gashi D, Tanaka T, et al.
    Improving high viscosity extrusion of microcrystals for time-resolved serial femtosecond crystallography at X-ray lasers
    Journal of Visualized Experiments. 2019;(144): e59087. https://doi.org/10.3791/59087
    DORA PSI
  • Mayer D, Damberger FF, Samarasimhareddy M, Feldmueller M, Vuckovic Z, Flock T, et al.
    Distinct G protein-coupled receptor phosphorylation motifs modulate arrestin affinity and activation and global conformation
    Nature Communications. 2019; 10: 1261 (14 pp.). https://doi.org/10.1038/s41467-019-09204-y
    DORA PSI
  • Standfuss J
    Membrane protein dynamics studied by X-ray lasers - or why only time will tell
    Current Opinion in Structural Biology. 2019; 57: 63-71. https://doi.org/10.1016/j.sbi.2019.02.001
    DORA PSI
  • Weinert T, Skopintsev P, James D, Dworkowski F, Panepucci E, Kekilli D, et al.
    Proton uptake mechanism in bacteriorhodopsin captured by serial synchrotron crystallography
    Science. 2019; 365(6448): 61-65. https://doi.org/10.1126/science.aaw8634
    DORA PSI
  • Wickstrand C, Nogly P, Nango E, Iwata S, Standfuss J, Neutze R
    Bacteriorhodopsin: structural insights revealed using X-ray lasers and synchrotron radiation
    Annual Review of Biochemistry. 2019; 88: 59-83. https://doi.org/10.1146/annurev-biochem-013118-111327
    DORA PSI
  • Mattle D, Kuhn B, Aebi J, Bedoucha M, Kekilli D, Grozinger N, et al.
    Ligand channel in pharmacologically stabilized rhodopsin
    Proceedings of the National Academy of Sciences of the United States of America PNAS. 2018; 115(14): 3640-3645. https://doi.org/10.1073/pnas.1718084115
    DORA PSI
  • Nogly P, Weinert T, James D, Carbajo S, Ozerov D, Furrer A, et al.
    Retinal isomerization in bacteriorhodopsin captured by a femtosecond x-ray laser
    Science. 2018; 361(6398): eaat0094 (7 pp.). https://doi.org/10.1126/science.aat0094
    DORA PSI
  • Tsai C-J, Pamula F, Nehmé R, Mühle J, Weinert T, Flock T, et al.
    Crystal structure of rhodopsin in complex with a mini-Go sheds light on the principles of G protein selectivity
    Science Advances. 2018; 4(9): aat7052 (9 pp.). https://doi.org/10.1126/sciadv.aat7052
    DORA PSI
  • Abela R, Beaud P, van Bokhoven JA, Chergui M, Feurer T, Haase J, et al.
    Perspective: opportunities for ultrafast science at SwissFEL
    Structural Dynamics. 2017; 4(6): 61602 (25 pp.). https://doi.org/10.1063/1.4997222
    DORA PSI
  • Standfuss J, Spence J
    Serial crystallography at synchrotrons and X-ray lasers
    IUCrJ. 2017; 4: 100-101. https://doi.org/10.1107/S2052252517001877
    DORA PSI
  • Tsai C-J, Standfuss J
    Structural biology: signalling under the microscope
    Nature. 2017; 546(7656): 36-37. https://doi.org/10.1038/nature22491
    DORA PSI
  • Weinert T, Olieric N, Cheng R, Brünle S, James D, Ozerov D, et al.
    Serial millisecond crystallography for routine room-temperature structure determination at synchrotrons
    Nature Communications. 2017; 8(1): 542 (11 pp.). https://doi.org/10.1038/s41467-017-00630-4
    DORA PSI
  • Jaeger K, Dworkowski F, Nogly P, Milne C, Wang M, Standfuss J
    Serial millisecond crystallography of membrane proteins
    In: Moraes I, ed. The next generation in membrane protein structure determination. Advances in experimental medicine and biology. Cham: Springer; 2016:137-149. https://doi.org/10.1007/978-3-319-35072-1_10
    DORA PSI
  • Nango E, Royant A, Kubo M, Nakane T, Wickstrand C, Kimura T, et al.
    A three-dimensional movie of structural changes in bacteriorhodopsin
    Science. 2016; 354(6319): 1552-1557. https://doi.org/10.1126/science.aaH3497
    DORA PSI
  • Nogly P, Panneels V, Nelson G, Gati C, Kimura T, Milne C, et al.
    Lipidic cubic phase injector is a viable crystal delivery system for time-resolved serial crystallography
    Nature Communications. 2016; 7: 12314 (9 pp.). https://doi.org/10.1038/ncomms12314
    DORA PSI
  • Peterhans C, Lally CCM, Ostermaier MK, Sommer ME, Standfuss J
    Functional map of arrestin binding to phosphorylated opsin, with and without agonist
    Scientific Reports. 2016; 6: 28686 (14 pp.). https://doi.org/10.1038/srep28686
    DORA PSI
  • Singhal A, Guo Y, Matkovic M, Schertler G, Deupi X, Yan ECY, et al.
    Structural role of the T94I rhodopsin mutation in congenital stationary night blindness
    EMBO Reports. 2016; 17(10): 1431-1440. https://doi.org/10.15252/embr.201642671
    DORA PSI
  • Kang Y, Zhou XE, Gao X, He Y, Liu W, Ishchenko A, et al.
    Crystal structure of rhodopsin bound to arrestin by femtosecond X-ray laser
    Nature. 2015; 523: 561-567. https://doi.org/10.1038/nature14656
    DORA PSI
  • Mattle D, Singhal A, Schmid G, Dawson R, Standfuss J
    Mammalian expression, purification, and crystallization of rhodopsin variants
    In: Jastrzebska B, ed. Rhodopsin. Methods and protocols. Methods in molecular biology. New York: Humana Press; 2015:39-54. https://doi.org/10.1007/978-1-4939-2330-4_3
    DORA PSI
  • Nogly P, Standfuss J
    Light-driven Na+ pumps as next-generation inhibitory optogenetic tools
    Nature Structural and Molecular Biology. 2015; 22(5): 351-353. https://doi.org/10.1038/nsmb.3017
    DORA PSI
  • Nogly P, James D, Wang D, White TA, Zatsepin N, Shilova A, et al.
    Lipidic cubic phase serial millisecond crystallography using synchrotron radiation
    IUCrJ. 2015; 2: 168-176. https://doi.org/10.1107/S2052252514026487
    DORA PSI
  • Panneels V, Wu W, Tsai C-J, Nogly P, Rheinberger J, Jaeger K, et al.
    Time-resolved structural studies with serial crystallography: a new light on retinal proteins
    Structural Dynamics. 2015; 2(4): 041718 (8 pp.). https://doi.org/10.1063/1.4922774
    DORA PSI
  • Standfuss J
    Viral chemokine mimicry. How do viruses trick the human immune system?
    Science. 2015; 347(6226): 1071-1072. https://doi.org/10.1126/science.aaa7998
    DORA PSI
  • Wu W, Nogly P, Rheinberger J, Kick LM, Gati C, Nelson G, et al.
    Batch crystallization of rhodopsin for structural dynamics using an X-ray free-electron laser
    Acta Crystallographica Section F: Structural Biology and Crystallization Communications. 2015; 71: 856-860. https://doi.org/10.1107/S2053230X15009966
    DORA PSI
  • Maeda S, Sun D, Singhal A, Foggetta M, Schmid G, Standfuss J, et al.
    Crystallization scale preparation of a stable GPCR signaling complex between constitutively active rhodopsin and G-protein
    PLoS One. 2014; 9(6): e98714 (11 pp.). https://doi.org/10.1371/journal.pone.0098714
    DORA PSI
  • Ostermaier MK, Peterhans C, Jaussi R, Deupi X, Standfuss J
    Functional map of arrestin-1 at single amino acid resolution
    Proceedings of the National Academy of Sciences of the United States of America PNAS. 2014; 111(5): 1825-1830. https://doi.org/10.1073/pnas.1319402111
    DORA PSI
  • Ostermaier MK, Schertler GFX, Standfuss J
    Molecular mechanism of phosphorylation-dependent arrestin activation
    Current Opinion in Structural Biology. 2014; 29: 143-151. https://doi.org/10.1016/j.sbi.2014.07.006
    DORA PSI
  • Brueckner F, Piscitelli CL, Tsai C-J, Standfuss J, Deupi X, Schertler GFX
    Structure of β-Adrenergic receptors
    In: Conn PM, ed. G protein coupled receptors. Structure. Methods in enzymology. San Diego: Elsevier; 2013:117-151. https://doi.org/10.1016/B978-0-12-391861-1.00006-X
    DORA PSI
  • Singhal A, Ostermaier MK, Vishnivetskiy SA, Panneels V, Homan KT, Tesmer JJG, et al.
    Insights into congenital stationary night blindness based on the structure of G90D rhodopsin
    EMBO Reports. 2013; 14(6): 520-526. https://doi.org/10.1038/embor.2013.44
    DORA PSI
  • Sun D, Ostermaier MK, Heydenreich FM, Mayer D, Jaussi R, Standfuss J, et al.
    AAscan, PCRdesign and MutantChecker: a suite of programs for primer design and sequence analysis for high-throughput scanning mutagenesis.
    PLoS One. 2013; 8(10): e78878 (9 pp.). https://doi.org/10.1371/journal.pone.0078878
    DORA PSI
  • Vishnivetskiy SA, Ostermaier MK, Singhal A, Panneels V, Homan KT, Glukhova A, et al.
    Constitutively active rhodopsin mutants causing night blindness are effectively phosphorylated by GRKs but differ in arrestin-1 binding
    Cellular Signalling. 2013; 25(11): 2155-2162. https://doi.org/10.1016/j.cellsig.2013.07.009
    DORA PSI
  • Deupi X, Standfuss J, Schertler G
    Conserved activation pathways in G-protein-coupled receptors
    Biochemical Society Transactions. 2012; 40(2): 383-388. https://doi.org/10.1042/BST20120001
    DORA PSI
  • Deupi X, Edwards P, Singhal A, Nickle B, Oprian D, Schertler G, et al.
    Stabilized G protein binding site in the structure of constitutively active metarhodopsin-II
    Proceedings of the National Academy of Sciences of the United States of America PNAS. 2012; 109(1): 119-124. https://doi.org/10.1073/pnas.1114089108
    DORA PSI
  • Deupi X, Standfuss J
    Structural insights into agonist-induced activation of G-protein-coupled receptors
    Current Opinion in Structural Biology. 2011; 21(4): 541-551. https://doi.org/10.1016/j.sbi.2011.06.002
    DORA PSI
  • Standfuss J, Edwards PC, D'Antona A, Fransen M, Xie G, Oprian DD, et al.
    The structural basis of agonist-induced activation in constitutively active rhodopsin
    Nature. 2011; 471(7340): 656-660. https://doi.org/10.1038/nature09795
    DORA PSI
  • Xie G, D'Antona AM, Edwards PC, Fransen M, Standfuss J, Schertler GFX, et al.
    Preparation of an activated rhodopsin/transducin complex using a constitutively active mutant of rhodopsin
    Biochemistry. 2011; 50(47): 10399-10407. https://doi.org/10.1021/bi201126r
    DORA PSI