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1. Structural effects of high laser power densities on an early bacteriorhodopsin photocycle intermediate

Author

Bertrand, Quentin, Nogly, Przemyslaw, Nango, Eriko, Kekilli, Demet, Khusainov, Georgii, Furrer, Antonia, James, Daniel, Dworkowski, Florian, Skopintsev, Petr, Mous, Sandra, Martiel, Isabelle, Börjesson, Per, Ortolani, Giorgia, Huang, Chia-Ying, Kepa, Michal, Ozerov, Dmitry, Brünle, Steffen, Panneels, Valerie, Tanaka, Tomoyuki, Tanaka, Rie, Tono, Kensuke, Owada, Shigeki, Johnson, Philip J. M., Nass, Karol, Knopp, Gregor, Cirelli, Claudio, Milne, Christopher, Schertler, Gebhard, Iwata, So, Neutze, Richard, Weinert, Tobias, and Standfuss, Jörg

Published
2024
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2. Structural effects of high laser power densities on an early bacteriorhodopsin photocycle intermediate

Author

Quentin Bertrand, Przemyslaw Nogly, Eriko Nango, Demet Kekilli, Georgii Khusainov, Antonia Furrer, Daniel James, Florian Dworkowski, Petr Skopintsev, Sandra Mous, Isabelle Martiel, Per Börjesson, Giorgia Ortolani, Chia-Ying Huang, Michal Kepa, Dmitry Ozerov, Steffen Brünle, Valerie Panneels, Tomoyuki Tanaka, Rie Tanaka, Kensuke Tono, Shigeki Owada, Philip J. M. Johnson, Karol Nass, Gregor Knopp, Claudio Cirelli, Christopher Milne, Gebhard Schertler, So Iwata, Richard Neutze, Tobias Weinert, and Jörg Standfuss

Subjects
Science
Abstract

Abstract Time-resolved serial crystallography at X-ray Free Electron Lasers offers the opportunity to observe ultrafast photochemical reactions at the atomic level. The technique has yielded exciting molecular insights into various biological processes including light sensing and photochemical energy conversion. However, to achieve sufficient levels of activation within an optically dense crystal, high laser power densities are often used, which has led to an ongoing debate to which extent photodamage may compromise interpretation of the results. Here we compare time-resolved serial crystallographic data of the bacteriorhodopsin K-intermediate collected at laser power densities ranging from 0.04 to 2493 GW/cm2 and follow energy dissipation of the absorbed photons logarithmically from picoseconds to milliseconds. Although the effects of high laser power densities on the overall structure are small, in the upper excitation range we observe significant changes in retinal conformation and increased heating of the functionally critical counterion cluster. We compare light-activation within crystals to that in solution and discuss the impact of the observed changes on bacteriorhodopsin biology.

Published
2024
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3. Capturing the blue-light activated state of the Phot-LOV1 domain from Chlamydomonas reinhardtii using time-resolved serial synchrotron crystallography

Author

Guillaume Gotthard, Sandra Mous, Tobias Weinert, Raiza Nara Antonelli Maia, Daniel James, Florian Dworkowski, Dardan Gashi, Antonia Furrer, Dmitry Ozerov, Ezequiel Panepucci, Meitian Wang, Gebhard F. X. Schertler, Joachim Heberle, Joerg Standfuss, and Przemyslaw Nogly

Subjects
time-resolved serial synchrotron crystallography, tr-ssx, room-temperature crystallography, blue-light photoreceptors, chlamydomonas reinhardtii, crphotlov1, structural dynamics, light–oxygen–voltage domains, Crystallography, QD901-999
Abstract

Light–oxygen–voltage (LOV) domains are small photosensory flavoprotein modules that allow the conversion of external stimuli (sunlight) into intracellular signals responsible for various cell behaviors (e.g. phototropism and chloroplast relocation). This ability relies on the light-induced formation of a covalent thioether adduct between a flavin chromophore and a reactive cysteine from the protein environment, which triggers a cascade of structural changes that result in the activation of a serine/threonine (Ser/Thr) kinase. Recent developments in time-resolved crystallography may allow the activation cascade of the LOV domain to be observed in real time, which has been elusive. In this study, we report a robust protocol for the production and stable delivery of microcrystals of the LOV domain of phototropin Phot-1 from Chlamydomonas reinhardtii (CrPhotLOV1) with a high-viscosity injector for time-resolved serial synchrotron crystallography (TR-SSX). The detailed process covers all aspects, from sample optimization to data collection, which may serve as a guide for soluble protein preparation for TR-SSX. In addition, we show that the crystals obtained preserve the photoreactivity using infrared spectroscopy. Furthermore, the results of the TR-SSX experiment provide high-resolution insights into structural alterations of CrPhotLOV1 from Δt = 2.5 ms up to Δt = 95 ms post-photoactivation, including resolving the geometry of the thioether adduct and the C-terminal region implicated in the signal transduction process.

Published
2024
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4. Fixed-target pump–probe SFX: eliminating the scourge of light contamination

Author

Guillaume Gotthard, Andrea Flores-Ibarra, Melissa Carrillo, Michal W. Kepa, Thomas J. Mason, Dennis P. Stegmann, Bence Olasz, Magdalena Pachota, Florian Dworkowski, Dmitry Ozerov, Bill F. Pedrini, Celestino Padeste, John H. Beale, and Przemyslaw Nogly

Subjects
time-resolved crystallography, fixed targets, x-ray free-electron lasers, room-temperature crystallography, pump–probe, photoreceptor light–oxygen–voltage domains, serial femtosecond crystallography, light contamination, sample consumption, Crystallography, QD901-999
Abstract

X-ray free-electron laser (XFEL) light sources have enabled the rapid growth of time-resolved structural experiments, which provide crucial information on the function of macromolecules and their mechanisms. Here, the aim was to commission the SwissMX fixed-target sample-delivery system at the SwissFEL Cristallina experimental station using the PSI-developed micro-structured polymer (MISP) chip for pump–probe time-resolved experiments. To characterize the system, crystals of the light-sensitive protein light–oxygen–voltage domain 1 (LOV1) from Chlamydomonas reinhardtii were used. Using different experimental settings, the accidental illumination, referred to as light contamination, of crystals mounted in wells adjacent to those illuminated by the pump laser was examined. It was crucial to control the light scattering from and through the solid supports otherwise significant contamination occurred. However, the results here show that the opaque MISP chips are suitable for defined pump–probe studies of a light-sensitive protein. The experiment also probed the sub-millisecond structural dynamics of LOV1 and indicated that at Δt = 10 µs a covalent thioether bond is established between reactive Cys57 and its flavin mononucleotide cofactor. This experiment validates the crystals to be suitable for in-depth follow-up studies of this still poorly understood signal-transduction mechanism. Importantly, the fixed-target delivery system also permitted a tenfold reduction in protein sample consumption compared with the more common high-viscosity extrusion-based delivery system. This development creates the prospect of an increase in XFEL project throughput for the field.

Published
2024
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5. Ultrafast structural changes direct the first molecular events of vision

Author

Gruhl, Thomas, Weinert, Tobias, Rodrigues, Matthew J., Milne, Christopher J., Ortolani, Giorgia, Nass, Karol, Nango, Eriko, Sen, Saumik, Johnson, Philip J. M., Cirelli, Claudio, Furrer, Antonia, Mous, Sandra, Skopintsev, Petr, James, Daniel, Dworkowski, Florian, Båth, Petra, Kekilli, Demet, Ozerov, Dmitry, Tanaka, Rie, Glover, Hannah, Bacellar, Camila, Brünle, Steffen, Casadei, Cecilia M., Diethelm, Azeglio D., Gashi, Dardan, Gotthard, Guillaume, Guixà-González, Ramon, Joti, Yasumasa, Kabanova, Victoria, Knopp, Gregor, Lesca, Elena, Ma, Pikyee, Martiel, Isabelle, Mühle, Jonas, Owada, Shigeki, Pamula, Filip, Sarabi, Daniel, Tejero, Oliver, Tsai, Ching-Ju, Varma, Niranjan, Wach, Anna, Boutet, Sébastien, Tono, Kensuke, Nogly, Przemyslaw, Deupi, Xavier, Iwata, So, Neutze, Richard, Standfuss, Jörg, Schertler, Gebhard, and Panneels, Valerie

Published
2023
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6. Lipidic cubic phase injector is a viable crystal delivery system for time-resolved serial crystallography

Author

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, Koglin, Jason E, Berntsen, Peter, Nango, Eriko, Iwata, So, Chapman, Henry N, Fromme, Petra, Frank, Matthias, Abela, Rafael, Boutet, Sébastien, Barty, Anton, White, Thomas A, Weierstall, Uwe, Spence, John, Neutze, Richard, Schertler, Gebhard, and Standfuss, Jörg

Subjects
Chemical Sciences, Physical Chemistry, Bioengineering, Nanotechnology, Bacteriorhodopsins, Crystallography, X-Ray, Feasibility Studies, Lasers, Lipids, Protein Conformation, Synchrotrons, Time Factors, Viscosity, X-Ray Absorption Spectroscopy
Abstract

Serial femtosecond crystallography (SFX) using X-ray free-electron laser sources is an emerging method with considerable potential for time-resolved pump-probe experiments. Here we present a lipidic cubic phase SFX structure of the light-driven proton pump bacteriorhodopsin (bR) to 2.3 Å resolution and a method to investigate protein dynamics with modest sample requirement. Time-resolved SFX (TR-SFX) with a pump-probe delay of 1 ms yields difference Fourier maps compatible with the dark to M state transition of bR. Importantly, the method is very sample efficient and reduces sample consumption to about 1 mg per collected time point. Accumulation of M intermediate within the crystal lattice is confirmed by time-resolved visible absorption spectroscopy. This study provides an important step towards characterizing the complete photocycle dynamics of retinal proteins and demonstrates the feasibility of a sample efficient viscous medium jet for TR-SFX.

Published
2016

7. Advances in long-wavelength native phasing at X-ray free-electron lasers

Author

Karol Nass, Robert Cheng, Laura Vera, Aldo Mozzanica, Sophie Redford, Dmitry Ozerov, Shibom Basu, Daniel James, Gregor Knopp, Claudio Cirelli, Isabelle Martiel, Cecilia Casadei, Tobias Weinert, Przemyslaw Nogly, Petr Skopintsev, Ivan Usov, Filip Leonarski, Tian Geng, Mathieu Rappas, Andrew S. Doré, Robert Cooke, Shahrooz Nasrollahi Shirazi, Florian Dworkowski, May Sharpe, Natacha Olieric, Camila Bacellar, Rok Bohinc, Michel O. Steinmetz, Gebhard Schertler, Rafael Abela, Luc Patthey, Bernd Schmitt, Michael Hennig, Jörg Standfuss, Meitian Wang, and Christopher J. Milne

Subjects
serial femtosecond crystallography, x-ray free-electron lasers, single-wavelength anomalous diffraction, de novo protein structure determination, anomalous data-quality indicators, Crystallography, QD901-999
Abstract

Long-wavelength pulses from the Swiss X-ray free-electron laser (XFEL) have been used for de novo protein structure determination by native single-wavelength anomalous diffraction (native-SAD) phasing of serial femtosecond crystallography (SFX) data. In this work, sensitive anomalous data-quality indicators and model proteins were used to quantify improvements in native-SAD at XFELs such as utilization of longer wavelengths, careful experimental geometry optimization, and better post-refinement and partiality correction. Compared with studies using shorter wavelengths at other XFELs and older software versions, up to one order of magnitude reduction in the required number of indexed images for native-SAD was achieved, hence lowering sample consumption and beam-time requirements significantly. Improved data quality and higher anomalous signal facilitate so-far underutilized de novo structure determination of challenging proteins at XFELs. Improvements presented in this work can be used in other types of SFX experiments that require accurate measurements of weak signals, for example time-resolved studies.

Published
2020
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8. Femtosecond-to-millisecond structural changes in a light-driven sodium pump

Author

Skopintsev, Petr, Ehrenberg, David, Weinert, Tobias, James, Daniel, Kar, Rajiv K., Johnson, Philip J. M., Ozerov, Dmitry, Furrer, Antonia, Martiel, Isabelle, Dworkowski, Florian, Nass, Karol, Knopp, Gregor, Cirelli, Claudio, Arrell, Christopher, Gashi, Dardan, Mous, Sandra, Wranik, Maximilian, Gruhl, Thomas, Kekilli, Demet, Brünle, Steffen, Deupi, Xavier, Schertler, Gebhard F. X., Benoit, Roger M., Panneels, Valerie, Nogly, Przemyslaw, Schapiro, Igor, Milne, Christopher, Heberle, Joachim, and Standfuss, Jörg

Published
2020
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9. A tool for visualizing protein motions in time-resolved crystallography

Author

Cecilia Wickstrand, Gergely Katona, Takanori Nakane, Przemyslaw Nogly, Joerg Standfuss, Eriko Nango, and Richard Neutze

Subjects
Crystallography, QD901-999
Abstract

Time-resolved serial femtosecond crystallography (TR-SFX) at an x-ray free electron laser enables protein structural changes to be imaged on time-scales from femtoseconds to seconds. It can, however, be difficult to grasp the nature and timescale of global protein motions when structural changes are not isolated near a single active site. New tools are, therefore, needed to represent the global nature of electron density changes and their correlation with modeled protein structural changes. Here, we use TR-SFX data from bacteriorhodopsin to develop and validate a method for quantifying time-dependent electron density changes and correlating them throughout the protein. We define a spherical volume of difference electron density about selected atoms, average separately the positive and negative electron difference densities within each volume, and walk this spherical volume through all atoms within the protein. By correlating the resulting difference electron density amplitudes with time, our approach facilitates an initial assessment of the number and timescale of structural intermediates and highlights quake-like motions on the sub-picosecond timescale. This tool also allows structural models to be compared with experimental data using theoretical difference electron density changes calculated from refined resting and photo-activated structures.

Published
2020
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10. Serial millisecond crystallography for routine room-temperature structure determination at synchrotrons

Author

Tobias Weinert, Natacha Olieric, Robert Cheng, Steffen Brünle, Daniel James, Dmitry Ozerov, Dardan Gashi, Laura Vera, May Marsh, Kathrin Jaeger, Florian Dworkowski, Ezequiel Panepucci, Shibom Basu, Petr Skopintsev, Andrew S. Doré, Tian Geng, Robert M. Cooke, Mengning Liang, Andrea E. Prota, Valerie Panneels, Przemyslaw Nogly, Ulrich Ermler, Gebhard Schertler, Michael Hennig, Michel O. Steinmetz, Meitian Wang, and Jörg Standfuss

Subjects
Science
Abstract

Serial crystallography was developed for protein crystal data collection with X-ray free-electron lasers. Here the authors present several examples which show that serial crystallography using high-viscosity injectors can also be routinely employed for room-temperature data collection at synchrotrons.

Published
2017
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11. Lipidic cubic phase injector is a viable crystal delivery system for time-resolved serial crystallography

Author

Przemyslaw Nogly, Valerie Panneels, Garrett Nelson, Cornelius Gati, Tetsunari Kimura, Christopher Milne, Despina Milathianaki, Minoru Kubo, Wenting Wu, Chelsie Conrad, Jesse Coe, Richard Bean, Yun Zhao, Petra Båth, Robert Dods, Rajiv Harimoorthy, Kenneth R. Beyerlein, Jan Rheinberger, Daniel James, Daniel DePonte, Chufeng Li, Leonardo Sala, Garth J. Williams, Mark S. Hunter, Jason E. Koglin, Peter Berntsen, Eriko Nango, So Iwata, Henry N. Chapman, Petra Fromme, Matthias Frank, Rafael Abela, Sébastien Boutet, Anton Barty, Thomas A. White, Uwe Weierstall, John Spence, Richard Neutze, Gebhard Schertler, and Jörg Standfuss

Subjects
Science
Abstract

Serial femtosecond crystallography using X-ray free-electron lasers has huge potential for time-resolved structural experiments. Here, the authors present a structure of the light-driven proton pump bacteriorhodopsin using these techniques.

Published
2016
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12. Crystal Structure of Mannose Specific IIA Subunit of Phosphotransferase System from Streptococcus pneumoniae

Author

Malgorzata Magoch, Przemyslaw Nogly, Przemyslaw Grudnik, Pikyee Ma, Bozena Boczkus, Ana Rute Neves, Margarida Archer, and Grzegorz Dubin

Subjects
Streptococcus pneumoniae, phosphotransferase system, IIA subunit, mannose PTS, sugar transport, X-ray crystallography, Organic chemistry, QD241-441
Abstract

Streptococcus pneumoniae is a frequent bacterial pathogen of the human respiratory tract causing pneumonia, meningitis and sepsis, a serious healthcare burden in all age groups. S. pneumoniae lacks complete respiratory chain and relies on carbohydrate fermentation for energy generation. One of the essential components for this includes the mannose phosphotransferase system (Man-PTS), which plays a central role in glucose transport and exhibits a broad specificity for a range of hexoses. Importantly, Man-PTS is involved in the global regulation of gene expression for virulence determinants. We herein report the three-dimensional structure of the EIIA domain of S. pneumoniae mannose phosphotransferase system (SpEIIA-Man). Our structure shows a dimeric arrangement of EIIA and reveals a detailed molecular description of the active site. Since PTS transporters are exclusively present in microbes and sugar transporters have already been suggested as valid targets for antistreptococcal antibiotics, our work sets foundation for the future development of antimicrobial strategies against Streptococcus pneumoniae.

Published
2020
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13. Lipidic cubic phase serial millisecond crystallography using synchrotron radiation

Author

Przemyslaw Nogly, Daniel James, Dingjie Wang, Thomas A. White, Nadia Zatsepin, Anastasya Shilova, Garrett Nelson, Haiguang Liu, Linda Johansson, Michael Heymann, Kathrin Jaeger, Markus Metz, Cecilia Wickstrand, Wenting Wu, Petra Båth, Peter Berntsen, Dominik Oberthuer, Valerie Panneels, Vadim Cherezov, Henry Chapman, Gebhard Schertler, Richard Neutze, John Spence, Isabel Moraes, Manfred Burghammer, Joerg Standfuss, and Uwe Weierstall

Subjects
lipidic cubic phases, protein crystallography, bacteriorhodopsin, XFEL, Crystallography, QD901-999
Abstract

Lipidic cubic phases (LCPs) have emerged as successful matrixes for the crystallization of membrane proteins. Moreover, the viscous LCP also provides a highly effective delivery medium for serial femtosecond crystallography (SFX) at X-ray free-electron lasers (XFELs). Here, the adaptation of this technology to perform serial millisecond crystallography (SMX) at more widely available synchrotron microfocus beamlines is described. Compared with conventional microcrystallography, LCP-SMX eliminates the need for difficult handling of individual crystals and allows for data collection at room temperature. The technology is demonstrated by solving a structure of the light-driven proton-pump bacteriorhodopsin (bR) at a resolution of 2.4 Å. The room-temperature structure of bR is very similar to previous cryogenic structures but shows small yet distinct differences in the retinal ligand and proton-transfer pathway.

Published
2015
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14. NmHR light state structure at 55 ms (50 - 60 ms) after photoexcitation determined by serial millisecond crystallography

Author

Mous, S., primary, Gotthard, G., additional, Ehrenberg, D., additional, Sen, S., additional, James, D., additional, Johnson, P., additional, Weinert, T., additional, Nass, K., additional, Furrer, A., additional, Kekilli, D., additional, Ma, P., additional, Bruenle, S., additional, Casadei, C., additional, Martiel, I., additional, Dworkowski, F., additional, Gashi, D., additional, Skopintsev, P., additional, Wranik, M., additional, Knopp, G., additional, Panepucci, E., additional, Panneels, V., additional, Cirelli, C., additional, Ozerov, D., additional, Schertler, G., additional, Wang, M., additional, Milne, C., additional, Standfuss, J., additional, Schapiro, I., additional, Heberle, J., additional, and Nogly, P., additional

Published
2022
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15. Anomalous bromide substructure of NmHR under dark state conditions determined at 13.7 keV with serial crystallography

Author

Mous, S., primary, Gotthard, G., additional, Ehrenberg, D., additional, Sen, S., additional, James, D., additional, Johnson, P., additional, Weinert, T., additional, Nass, K., additional, Furrer, A., additional, Kekilli, D., additional, Ma, P., additional, Bruenle, S., additional, Casadei, C., additional, Martiel, I., additional, Dworkowski, F., additional, Gashi, D., additional, Skopintsev, P., additional, Wranik, M., additional, Knopp, G., additional, Panepucci, E., additional, Panneels, V., additional, Cirelli, C., additional, Ozerov, D., additional, Schertler, G., additional, Wang, M., additional, Milne, C., additional, Standfuss, J., additional, Schapiro, I., additional, Heberle, J., additional, and Nogly, P., additional

Published
2022
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16. Time-resolved structural studies with serial crystallography: A new light on retinal proteins

Author

Valérie Panneels, Wenting Wu, Ching-Ju Tsai, Przemek Nogly, Jan Rheinberger, Kathrin Jaeger, Gregor Cicchetti, Cornelius Gati, Leonhard M. Kick, Leonardo Sala, Guido Capitani, Chris Milne, Celestino Padeste, Bill Pedrini, Xiao-Dan Li, Jörg Standfuss, Rafael Abela, and Gebhard Schertler

Subjects
Crystallography, QD901-999
Abstract

Structural information of the different conformational states of the two prototypical light-sensitive membrane proteins, bacteriorhodopsin and rhodopsin, has been obtained in the past by X-ray cryo-crystallography and cryo-electron microscopy. However, these methods do not allow for the structure determination of most intermediate conformations. Recently, the potential of X-Ray Free Electron Lasers (X-FELs) for tracking the dynamics of light-triggered processes by pump-probe serial femtosecond crystallography has been demonstrated using 3D-micron-sized crystals. In addition, X-FELs provide new opportunities for protein 2D-crystal diffraction, which would allow to observe the course of conformational changes of membrane proteins in a close-to-physiological lipid bilayer environment. Here, we describe the strategies towards structural dynamic studies of retinal proteins at room temperature, using injector or fixed-target based serial femtosecond crystallography at X-FELs. Thanks to recent progress especially in sample delivery methods, serial crystallography is now also feasible at synchrotron X-ray sources, thus expanding the possibilities for time-resolved structure determination.

Published
2015
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18. An efficient strategy for small-scale screening and production of archaeal membrane transport proteins in Escherichia coli.

Author

Pikyee Ma, Filipa Varela, Malgorzata Magoch, Ana Rita Silva, Ana Lúcia Rosário, José Brito, Tânia Filipa Oliveira, Przemyslaw Nogly, Miguel Pessanha, Meike Stelter, Arnulf Kletzin, Peter J F Henderson, and Margarida Archer

Subjects
Medicine, Science
Abstract

Membrane proteins play a key role in many fundamental cellular processes such as transport of nutrients, sensing of environmental signals and energy transduction, and account for over 50% of all known drug targets. Despite their importance, structural and functional characterisation of membrane proteins still remains a challenge, partially due to the difficulties in recombinant expression and purification. Therefore the need for development of efficient methods for heterologous production is essential.Fifteen integral membrane transport proteins from Archaea were selected as test targets, chosen to represent two superfamilies widespread in all organisms known as the Major Facilitator Superfamily (MFS) and the 5-Helix Inverted Repeat Transporter superfamily (5HIRT). These proteins typically have eleven to twelve predicted transmembrane helices and are putative transporters for sugar, metabolite, nucleobase, vitamin or neurotransmitter. They include a wide range of examples from the following families: Metabolite-H(+)-symporter; Sugar Porter; Nucleobase-Cation-Symporter-1; Nucleobase-Cation-Symporter-2; and neurotransmitter-sodium-symporter. Overproduction of transporters was evaluated with three vectors (pTTQ18, pET52b, pWarf) and two Escherichia coli strains (BL21 Star and C43 (DE3)). Thirteen transporter genes were successfully expressed; only two did not express in any of the tested vector-strain combinations. Initial trials showed that seven transporters could be purified and six of these yielded quantities of ≥ 0.4 mg per litre suitable for functional and structural studies. Size-exclusion chromatography confirmed that two purified transporters were almost homogeneous while four others were shown to be non-aggregating, indicating that they are ready for up-scale production and crystallisation trials.Here, we describe an efficient strategy for heterologous production of membrane transport proteins in E. coli. Small-volume cultures (10 mL) produced sufficient amount of proteins to assess their purity and aggregation state. The methods described in this work are simple to implement and can be easily applied to many more membrane proteins.

Published
2013
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20. Structure of thaumatin determined at SwissFEL using native-SAD at 4.57 keV from 20,000 diffraction patterns

Author

Nass, K., primary, Cheng, R., additional, Vera, L., additional, Mozzanica, A., additional, Redford, S., additional, Ozerov, D., additional, Basu, S., additional, James, D., additional, Knopp, G., additional, Cirelli, C., additional, Martiel, I., additional, Casadei, C., additional, Weinert, T., additional, Nogly, P., additional, Skopintsev, P., additional, Usov, I., additional, Leonarski, F., additional, Geng, T., additional, Rappas, M., additional, Dore, A.S., additional, Cooke, R., additional, Nasrollahi Shirazi, S., additional, Dworkowski, F., additional, Sharpe, M., additional, Olieric, N., additional, Steinmetz, M.O., additional, Schertler, G., additional, Abela, R., additional, Patthey, L., additional, Schmitt, B., additional, Hennig, M., additional, Standfuss, J., additional, Wang, M., additional, and Milne, J.C., additional

Published
2020
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21. Structure of thaumatin determined at SwissFEL using native-SAD at 6.06 keV from 50,000 diffraction patterns.

Author

Nass, K., primary, Cheng, R., additional, Vera, L., additional, Mozzanica, A., additional, Redford, S., additional, Ozerov, D., additional, Basu, S., additional, James, D., additional, Knopp, G., additional, Cirelli, C., additional, Martiel, I., additional, Casadei, C., additional, Weinert, T., additional, Nogly, P., additional, Skopintsev, P., additional, Usov, I., additional, Leonarski, F., additional, Geng, T., additional, Rappas, M., additional, Dore, A.S., additional, Cooke, R., additional, Nasrollahi Shirazi, S., additional, Dworkowski, F., additional, Sharpe, M., additional, Olieric, N., additional, Steinmetz, M.O., additional, Schertler, G., additional, Abela, R., additional, Patthey, L., additional, Schmitt, B., additional, Hennig, M., additional, Standfuss, J., additional, Wang, M., additional, and Milne, J.C., additional

Published
2020
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22. Structure of thaumatin determined at SwissFEL using native-SAD at 6.06 keV from all available diffraction patterns

Author

Nass, K., primary, Cheng, R., additional, Vera, L., additional, Mozzanica, A., additional, Redford, S., additional, Ozerov, D., additional, Basu, S., additional, James, D., additional, Knopp, G., additional, Cirelli, C., additional, Martiel, I., additional, Casadei, C., additional, Weinert, T., additional, Nogly, P., additional, Skopintsev, P., additional, Usov, I., additional, Leonarski, F., additional, Geng, T., additional, Rappas, M., additional, Dore, A.S., additional, Cooke, R., additional, Nasrollahi Shirazi, S., additional, Dworkowski, F., additional, Sharpe, M., additional, Olieric, N., additional, Steinmetz, M.O., additional, Schertler, G., additional, Abela, R., additional, Patthey, L., additional, Schmitt, B., additional, Hennig, M., additional, Standfuss, J., additional, Wang, M., additional, and Milne, J.C., additional

Published
2020
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23. Structure of the A2A adenosine receptor determined at SwissFEL using native-SAD at 4.57 keV from all available diffraction patterns

Author

Nass, K., primary, Cheng, R., additional, Vera, L., additional, Mozzanica, A., additional, Redford, S., additional, Ozerov, D., additional, Basu, S., additional, James, D., additional, Knopp, G., additional, Cirelli, C., additional, Martiel, I., additional, Casadei, C., additional, Weinert, T., additional, Nogly, P., additional, Skopintsev, P., additional, Usov, I., additional, Leonarski, F., additional, Geng, T., additional, Rappas, M., additional, Dore, A.S., additional, Cooke, R., additional, Nasrollahi Shirazi, S., additional, Dworkowski, F., additional, Sharpe, M., additional, Olieric, N., additional, Steinmetz, M.O., additional, Schertler, G., additional, Abela, R., additional, Patthey, L., additional, Schmitt, B., additional, Hennig, M., additional, Standfuss, J., additional, Wang, M., additional, and Milne, J.C., additional

Published
2020
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24. Structure of thaumatin determined at SwissFEL using native-SAD at 4.57 keV from all available diffraction patterns

Author

Nass, K., primary, Cheng, R., additional, Vera, L., additional, Mozzanica, A., additional, Redford, S., additional, Ozerov, D., additional, Basu, S., additional, James, D., additional, Knopp, G., additional, Cirelli, C., additional, Martiel, I., additional, Casadei, C., additional, Weinert, T., additional, Nogly, P., additional, Skopintsev, P., additional, Usov, I., additional, Leonarski, F., additional, Geng, T., additional, Rappas, M., additional, Dore, A.S., additional, Cooke, R., additional, Nasrollahi Shirazi, S., additional, Dworkowski, F., additional, Sharpe, M., additional, Olieric, N., additional, Steinmetz, M.O., additional, Schertler, G., additional, Abela, R., additional, Patthey, L., additional, Schmitt, B., additional, Hennig, M., additional, Standfuss, J., additional, Wang, M., additional, and Milne, J.Ch., additional

Published
2020
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25. Structure of the A2A adenosine receptor determined at SwissFEL using native-SAD at 4.57 keV from 50,000 diffraction patterns

Author

Nass, K., primary, Cheng, R., additional, Vera, L., additional, Mozzanica, A., additional, Redford, S., additional, Ozerov, D., additional, Basu, S., additional, James, D., additional, Knopp, G., additional, Cirelli, C., additional, Martiel, I., additional, Casadei, C., additional, Weinert, T., additional, Nogly, P., additional, Skopintsev, P., additional, Usov, I., additional, Leonarski, F., additional, Geng, T., additional, Rappas, M., additional, Dore, A.S., additional, Cooke, R., additional, Nasrollahi Shirazi, S., additional, Dworkowski, F., additional, Sharpe, M., additional, Olieric, N., additional, Steinmetz, M.O., additional, Schertler, G., additional, Abela, R., additional, Patthey, L., additional, Schmitt, B., additional, Hennig, M., additional, Standfuss, J., additional, Wang, M., additional, and Milne, J.C., additional

Published
2020
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26. Femtosecond to millisecond structural changes in a light-driven sodium pump: 1ns+16ns structure of KR2 with extrapolated, light and dark datasets

Author

Skopintsev, P., primary, Ehrenberg, D., additional, Weinert, T., additional, James, D., additional, Kar, R., additional, Johnson, P., additional, Ozerov, D., additional, Furrer, A., additional, Martiel, I., additional, Dworkowski, F., additional, Nass, K., additional, Knopp, G., additional, Cirelli, C., additional, Gashi, D., additional, Mous, S., additional, Wranik, M., additional, Gruhl, T., additional, Kekilli, D., additional, Bruenle, S., additional, Deupi, X., additional, Schertler, G.F.X., additional, Benoit, R., additional, Panneels, V., additional, Nogly, P., additional, Schapiro, I., additional, Milne, C., additional, Heberle, J., additional, and Standfuss, J., additional

Published
2020
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27. Femtosecond to millisecond structural changes in a light-driven sodium pump: Dark structure in acidic conditions

Author

Skopintsev, P., primary, Ehrenberg, D., additional, Weinert, T., additional, James, D., additional, Kar, R., additional, Johnson, P., additional, Ozerov, D., additional, Furrer, A., additional, Martiel, I., additional, Dworkowski, F., additional, Nass, K., additional, Knopp, G., additional, Cirelli, C., additional, Gashi, D., additional, Mous, S., additional, Wranik, M., additional, Gruhl, T., additional, Kekilli, D., additional, Bruenle, S., additional, Deupi, X., additional, Schertler, G.F.X., additional, Benoit, R., additional, Panneels, V., additional, Nogly, P., additional, Schapiro, I., additional, Milne, C., additional, Heberle, J., additional, and Standfuss, J., additional

Published
2020
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28. Femtosecond to millisecond structural changes in a light-driven sodium pump: 30us+150us structure of KR2 with extrapolated, light and dark datasets

Author

Skopintsev, P., primary, Ehrenberg, D., additional, Weinert, T., additional, James, D., additional, Kar, R., additional, Johnson, P., additional, Ozerov, D., additional, Furrer, A., additional, Martiel, I., additional, Dworkowski, F., additional, Nass, K., additional, Knopp, G., additional, Cirelli, C., additional, Gashi, D., additional, Mous, S., additional, Wranik, M., additional, Gruhl, T., additional, Kekilli, D., additional, Bruenle, S., additional, Deupi, X., additional, Schertler, G.F.X., additional, Benoit, R., additional, Panneels, V., additional, Nogly, P., additional, Schapiro, I., additional, Milne, C., additional, Heberle, J., additional, and Standfuss, J., additional

Published
2020
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29. Femtosecond to millisecond structural changes in a light-driven sodium pump: 800fs+2ps structure of KR2 with extrapolated, light and dark datasets

Author

Skopintsev, P., primary, Ehrenberg, D., additional, Weinert, T., additional, James, D., additional, Kar, R., additional, Johnson, P., additional, Ozerov, D., additional, Furrer, A., additional, Martiel, I., additional, Dworkowski, F., additional, Nass, K., additional, Knopp, G., additional, Cirelli, C., additional, Gashi, D., additional, Mous, S., additional, Wranik, M., additional, Gruhl, T., additional, Kekilli, D., additional, Bruenle, S., additional, Deupi, X., additional, Schertler, G.F.X., additional, Benoit, R., additional, Panneels, V., additional, Nogly, P., additional, Schapiro, I., additional, Milne, C., additional, Heberle, J., additional, and Standfuss, J., additional

Published
2020
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30. Femtosecond to millisecond structural changes in a light-driven sodium pump: 20ms structure of KR2 with extrapolated, light and dark datasets

Author

Skopintsev, P., primary, Ehrenberg, D., additional, Weinert, T., additional, James, D., additional, Kar, R., additional, Johnson, P., additional, Ozerov, D., additional, Furrer, A., additional, Martiel, I., additional, Dworkowski, F., additional, Nass, K., additional, Knopp, G., additional, Cirelli, C., additional, Gashi, D., additional, Mous, S., additional, Wranik, M., additional, Gruhl, T., additional, Kekilli, D., additional, Bruenle, S., additional, Deupi, X., additional, Schertler, G.F.X., additional, Benoit, R., additional, Panneels, V., additional, Nogly, P., additional, Schapiro, I., additional, Milne, C., additional, Heberle, J., additional, and Standfuss, J., additional

Published
2020
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31. Femtosecond to millisecond structural changes in a light-driven sodium pump: Dark structure in neutral conditions with attached light datasets at 800fs, 2ps, 100ps, 1ns, 16ns, 1us, 30us, 150us, 1ms and 20ms

Author

Skopintsev, P., primary, Ehrenberg, D., additional, Weinert, T., additional, James, D., additional, Kar, R., additional, Johnson, P., additional, Ozerov, D., additional, Furrer, A., additional, Martiel, I., additional, Dworkowski, F., additional, Nass, K., additional, Knopp, G., additional, Cirelli, C., additional, Gashi, D., additional, Mous, S., additional, Wranik, M., additional, Gruhl, T., additional, Kekilli, D., additional, Bruenle, S., additional, Deupi, X., additional, Schertler, G.F.X., additional, Benoit, R., additional, Panneels, V., additional, Nogly, P., additional, Schapiro, I., additional, Milne, C., additional, Heberle, J., additional, and Standfuss, J., additional

Published
2020
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32. Structure of the human CC Chemokine Receptor 7 in complex with the intracellular allosteric antagonist Cmp2105 and the insertion protein Sialidase NanA

Author

Jaeger, K., primary, Bruenle, S., additional, Weinert, T., additional, Guba, W., additional, Muehle, J., additional, Miyazaki, T., additional, Weber, M., additional, Furrer, A., additional, Haenggi, N., additional, Tetaz, T., additional, Huang, C.Y., additional, Mattle, D., additional, Vonach, J.M., additional, Gast, A., additional, Kuglstatter, A., additional, Rudolph, M.G., additional, Nogly, P., additional, Benz, J., additional, Dawson, R.J.P., additional, and Standfuss, J., additional

Published
2019
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38. Crystal structure of the rhodopsin-mini-Go complex

Author

Tsai, C.-J., primary, Weinert, T., additional, Muehle, J., additional, Pamula, F., additional, Nehme, R., additional, Flock, T., additional, Nogly, P., additional, Edwards, P.C., additional, Carpenter, B., additional, Gruhl, T., additional, Ma, P., additional, Deupi, X., additional, Standfuss, J., additional, Tate, C.G., additional, and Schertler, G.F.X., additional

Published
2018
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39. Retinal isomerization in bacteriorhodopsin revealed by a femtosecond X-ray laser: resting state structure

Author

Nogly, P., primary, Weinert, T., additional, James, D., additional, Cabajo, S., additional, Ozerov, D., additional, Furrer, A., additional, Gashi, D., additional, Borin, V., additional, Skopintsev, P., additional, Jaeger, K., additional, Nass, K., additional, Bath, P., additional, Bosman, R., additional, Koglin, J., additional, Seaberg, M., additional, Lane, T., additional, Kekilli, D., additional, Bruenle, S., additional, Tanaka, T., additional, Wu, W., additional, Milne, C., additional, White, T., additional, Barty, A., additional, Weierstall, U., additional, Panneels, V., additional, Nango, E., additional, Iwata, S., additional, Hunter, M., additional, Schapiro, I., additional, Schertler, G., additional, Neutze, R., additional, and Standfuss, J., additional

Published
2018
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40. Retinal isomerization in bacteriorhodopsin revealed by a femtosecond X-ray laser: 49-406 fs state structure

Author

Nogly, P., primary, Weinert, T., additional, James, D., additional, Cabajo, S., additional, Ozerov, D., additional, Furrer, A., additional, Gashi, D., additional, Borin, V., additional, Skopintsev, P., additional, Jaeger, K., additional, Nass, K., additional, Bath, P., additional, Bosman, R., additional, Koglin, J., additional, Seaberg, M., additional, Lane, T., additional, Kekilli, D., additional, Bruenle, S., additional, Tanaka, T., additional, Wu, W., additional, Milne, C., additional, White, T., additional, Barty, A., additional, Weierstall, U., additional, Panneels, V., additional, Nango, E., additional, Iwata, S., additional, Hunter, M., additional, Schapiro, I., additional, Schertler, G., additional, Neutze, R., additional, and Standfuss, J., additional

Published
2018
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41. Retinal isomerization in bacteriorhodopsin revealed by a femtosecond X-ray laser: 457-646 fs state structure

Author

Nogly, P., primary, Weinert, T., additional, James, D., additional, Cabajo, S., additional, Ozerov, D., additional, Furrer, A., additional, Gashi, D., additional, Borin, V., additional, Skopintsev, P., additional, Jaeger, K., additional, Nass, K., additional, Bath, P., additional, Bosman, R., additional, Koglin, J., additional, Seaberg, M., additional, Lane, T., additional, Kekilli, D., additional, Bruenle, S., additional, Tanaka, T., additional, Wu, W., additional, Milne, C., additional, White, T., additional, Barty, A., additional, Weierstall, U., additional, Panneels, V., additional, Nango, E., additional, Iwata, S., additional, Hunter, M., additional, Schapiro, I., additional, Schertler, G., additional, Neutze, R., additional, and Standfuss, J., additional

Published
2018
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42. Retinal isomerization in bacteriorhodopsin revealed by a femtosecond X-ray laser: 8.3 ms state structure

Author

Nogly, P., primary, Weinert, T., additional, James, D., additional, Cabajo, S., additional, Ozerov, D., additional, Furrer, A., additional, Gashi, D., additional, Borin, V., additional, Skopintsev, P., additional, Jaeger, K., additional, Nass, K., additional, Bath, P., additional, Bosman, R., additional, Koglin, J., additional, Seaberg, M., additional, Lane, T., additional, Kekilli, D., additional, Bruenle, S., additional, Tanaka, T., additional, Wu, W., additional, Milne, C., additional, White, T., additional, Barty, A., additional, Weierstall, U., additional, Panneels, V., additional, Nango, E., additional, Iwata, S., additional, Hunter, M., additional, Schapiro, I., additional, Schertler, G., additional, Neutze, R., additional, and Standfuss, J., additional

Published
2018
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43. Retinal isomerization in bacteriorhodopsin revealed by a femtosecond X-ray laser: 10 ps state structure

Author

Nogly, P., primary, Weinert, T., additional, James, D., additional, Cabajo, S., additional, Ozerov, D., additional, Furrer, A., additional, Gashi, D., additional, Borin, V., additional, Skopintsev, P., additional, Jaeger, K., additional, Nass, K., additional, Bath, P., additional, Bosman, R., additional, Koglin, J., additional, Seaberg, M., additional, Lane, T., additional, Kekilli, D., additional, Bruenle, S., additional, Tanaka, T., additional, Wu, W., additional, Milne, C., additional, White, T., additional, Barty, A., additional, Weierstall, U., additional, Panneels, V., additional, Nango, E., additional, Iwata, S., additional, Hunter, M., additional, Schapiro, I., additional, Schertler, G., additional, Neutze, R., additional, and Standfuss, J., additional

Published
2018
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45. Tubulin Darpin cryo structure

Author

Weinert, T., primary, Olieric, N., additional, James, D., additional, Gashi, D., additional, Nogly, P., additional, Jaeger, K., additional, Steinmetz, M.O., additional, and Standfuss, J., additional

Published
2017
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48. A2A Adenosine receptor cryo structure

Author

Weinert, T., primary, Cheng, R., additional, James, D., additional, Gashi, D., additional, Nogly, P., additional, Jaeger, K., additional, Dore, A.S., additional, Geng, T., additional, Cooke, R., additional, Hennig, M., additional, and Standfuss, J., additional

Published
2017
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