retinal binding proteins; light-driven pumps; serial femtosecond crystallography; optogenetics; G protein coupled receptors; visual photoreceptor rhodopsin; seven transmembrane proteins; kinetic crystallography; light-gated ion channels
Standfuss Jörg (2019), Membrane protein dynamics studied by X-ray lasers – or why only time will tell, in Current Opinion in Structural Biology
, 57, 63-71.
Weinert Tobias, Skopintsev Petr, James Daniel, Dworkowski Florian, Panepucci Ezequiel, Kekilli Demet, Furrer Antonia, Brünle Steffen, Mous Sandra, Ozerov Dmitry, Nogly Przemyslaw, Wang Meitian, Standfuss Jörg (2019), Proton uptake mechanism in bacteriorhodopsin captured by serial synchrotron crystallography, in Science
, 365(6448), 61-65.
Wickstrand Cecilia, Nogly Przemyslaw, Nango Eriko, Iwata So, Standfuss Jörg, Neutze Richard (2019), Bacteriorhodopsin: Structural Insights Revealed Using X-Ray Lasers and Synchrotron Radiation, in Annual Review of Biochemistry
, 88(1), 59-83.
James Daniel, Weinert Tobias, Skopintsev Petr, Furrer Antonia, Gashi Dardan, Tanaka Tomoyuki, Nango Eriko, Nogly Przemyslaw, Standfuss Joerg (2019), Improving High Viscosity Extrusion of Microcrystals for Time-resolved Serial Femtosecond Crystallography at X-ray Lasers, in Journal of Visualized Experiments
, (144), e59087.
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
In 2017 the Swiss X-ray Free Electron Laser (SwissFEL) will start operation at the Paul Scherrer Institute (PSI). One of the most promising applications of the X-ray free electron laser (XFEL) technology is the determination of molecular movies of protein function by time-resolved serial femtosecond crystallography (TR-SFX). Together structural and dynamic information will provide unique insights into the function of proteins as the principal building blocks of our biology. With its unique infrastructure within Switzerland, the PSI is in the ideal position to establish itself as a world leader in the study of protein structural dynamics. It already provides well-equipped laboratory space, robotic crystallization facilities and three world-class protein crystallography beamlines at the Swiss Light Source (SLS) to a large community of structural biologists. At the SLS nearly 5000 protein structures have been determined and deposited in public databases with many more resulting from work by private pharma companies and not publically available. Soon the PSI will extend this portfolio of services by giving access to SwissFEL to further expand its large national and international user base. Serial crystallography relying on the ultrashort, high brilliance X-ray pulses produced by SwissFEL will allow researchers to perform both radiation damage free room-temperature protein structure determinations and time-resolved measurements with femtosecond time resolution.In the last two years, we have established methods for routine room temperature serial crystallography experiments at both 3rd-generation storage rings and at several XFEL facilities worldwide. Here we propose to build on these experiences and install identical tunable nanosecond laser pump systems at SLS and SwissFEL that are optimized for time-resolved measurements on protein crystals. This will allow users to perform many different types of experiments on a wide range of timescales, all using identical excitation conditions, which is important for comparing data sets between experiments and facilities. Currently the available time resolution at the SLS is in the millisecond regime, which is primarily due to the limited diffraction power of protein crystals. The planned SLS-2 upgrade to a diffraction limited storage ring will dramatically improve the flux density of the X-ray beam and allow measurements to be performed on faster timescales and smaller crystals. Serial sample injection at the SLS will allow us to solve the active state of many proteins and can act as important test for users if more extensive pump probe studies can be done efficiently at SwissFEL. The ability to use nanosecond long laser pulses to efficiently photoexcite samples at SwissFEL will enable researchers to probe long-lived intermediates in the protein’s photocycle in the nanosecond to millisecond regime. This feature is complementary to the current infrastructure available at SwissFEL to perform femtosecond to picosecond time-resolved experiments. By this the envisaged setup will maximize synergy between the two advanced X-ray sources and allow users to choose the one most appropriate for their biological questions.