scholarly journals Investigating photoreception and bio-catalysis using time-resolved serial femtosecond crystallography

2021 ◽  
Vol 77 (a1) ◽  
pp. a21-a21
Author(s):  
Suraj Pandey ◽  
Marius Schmidt
Keyword(s):  
Time Resolved ◽  
Serial Femtosecond Crystallography

Time-Resolved Serial Femtosecond Crystallography at the European X-ray Free Electron Laser

2019 ◽  
Author(s):  
Marius Schmidt ◽  
Suraj Pandey ◽  
Adrian Mancuso ◽  
Richard Bean
Keyword(s):  
Free Electron ◽  
Free Electron Laser ◽  
Crystallographic Data ◽  
Time Resolved ◽  
X Ray ◽  
Serial Femtosecond Crystallography

Abstract This protocol introduces step by step into the collection of time resolved crystallographic data and their analysis at the European Free Electron Laser.

scholarly journals Time-resolved serial femtosecond crystallography at the European XFEL

2019 ◽  
Vol 17 (1) ◽  
pp. 73-78 ◽  
Author(s):  
Suraj Pandey ◽  
Richard Bean ◽  
Tokushi Sato ◽  
Ishwor Poudyal ◽  
Johan Bielecki ◽  
...  
Keyword(s):  
Time Resolved ◽  
Serial Femtosecond Crystallography

scholarly journals Improving High Viscosity Extrusion of Microcrystals for Time-resolved Serial Femtosecond Crystallography at X-ray Lasers

10.3791/59087 ◽  
2019 ◽  
Author(s):  
Daniel James ◽  
Tobias Weinert ◽  
Petr Skopintsev ◽  
Antonia Furrer ◽  
Dardan Gashi ◽  
...  
Keyword(s):  
High Viscosity ◽  
Time Resolved ◽  
X Ray ◽  
Serial Femtosecond Crystallography

scholarly journals Pump-Probe Time-Resolved Serial Femtosecond Crystallography at SACLA: Current Status and Data Collection Strategies

2019 ◽  
Vol 9 (24) ◽  
pp. 5505 ◽  
Author(s):  
Eriko Nango ◽  
Minoru Kubo ◽  
Kensuke Tono ◽  
So Iwata
Keyword(s):  
Data Collection ◽  
Free Electron ◽  
Free Electron Laser ◽  
Current Status ◽  
Optical Pump ◽  
Time Resolved ◽  
Pump Probe ◽  
X Ray ◽  
Collection Strategies ◽  
Serial Femtosecond Crystallography

Structural information on protein dynamics is a critical factor in fully understanding the protein functions. Pump-probe time-resolved serial femtosecond crystallography (TR-SFX) is a recently established technique for visualizing the structural changes or reactions in proteins that are at work with high spatial and temporal resolution. In the pump-probe method, protein microcrystals are continuously delivered from an injector and exposed to an X-ray free-electron laser (XFEL) pulse after a trigger to initiate a reaction, such as light, chemicals, temperature, and electric field, which affords the structural snapshots of intermediates that occur in the protein. We are in the process of developing the device and techniques for pump-probe TR-SFX while using XFEL produced at SPring-8 Angstrom Compact Free-Electron Laser (SACLA). In this paper, we described our current development details and data collection strategies for the optical pump X-ray probe TR-SFX experiment at SACLA and then reported the techniques of in crystallo TR spectroscopy, which is useful in clarifying the nature of reaction that takes place in crystals in advance.

scholarly journals Nanosecond pump–probe device for time-resolved serial femtosecond crystallography developed at SACLA

2017 ◽  
Vol 24 (5) ◽  
pp. 1086-1091 ◽  
Author(s):  
Minoru Kubo ◽  
Eriko Nango ◽  
Kensuke Tono ◽  
Tetsunari Kimura ◽  
Shigeki Owada ◽  
...  
Keyword(s):  
Conformational Changes ◽  
Free Electron ◽  
Good Choice ◽  
Cubic Phase ◽  
Liquid Droplets ◽  
Optical Pump ◽  
Time Resolved ◽  
X Ray ◽  
X Ray Crystallography ◽  
Serial Femtosecond Crystallography

X-ray free-electron lasers (XFELs) have opened new opportunities for time-resolved X-ray crystallography. Here a nanosecond optical-pump XFEL-probe device developed for time-resolved serial femtosecond crystallography (TR-SFX) studies of photo-induced reactions in proteins at the SPring-8 Angstrom Compact free-electron LAser (SACLA) is reported. The optical-fiber-based system is a good choice for a quick setup in a limited beam time and allows pump illumination from two directions to achieve high excitation efficiency of protein microcrystals. Two types of injectors are used: one for extruding highly viscous samples such as lipidic cubic phase (LCP) and the other for pulsed liquid droplets. Under standard sample flow conditions from the viscous-sample injector, delay times from nanoseconds to tens of milliseconds are accessible, typical time scales required to study large protein conformational changes. A first demonstration of a TR-SFX experiment on bacteriorhodopsin in bicelle using a setup with a droplet-type injector is also presented.

scholarly journals Time-resolved serial crystallography captures high-resolution intermediates of photoactive yellow protein

Science ◽  
2014 ◽  
Vol 346 (6214) ◽  
pp. 1242-1246 ◽  
Author(s):  
Jason Tenboer ◽  
Shibom Basu ◽  
Nadia Zatsepin ◽  
Kanupriya Pande ◽  
Despina Milathianaki ◽  
...  
Keyword(s):  
High Resolution ◽  
Protein A ◽  
Ultrashort Pulses ◽  
Photoactive Yellow Protein ◽  
Time Resolved ◽  
Density Maps ◽  
Serial Crystallography ◽  
Serial Femtosecond Crystallography ◽  
Reaction Initiation

Serial femtosecond crystallography using ultrashort pulses from x-ray free electron lasers (XFELs) enables studies of the light-triggered dynamics of biomolecules. We used microcrystals of photoactive yellow protein (a bacterial blue light photoreceptor) as a model system and obtained high-resolution, time-resolved difference electron density maps of excellent quality with strong features; these allowed the determination of structures of reaction intermediates to a resolution of 1.6 angstroms. Our results open the way to the study of reversible and nonreversible biological reactions on time scales as short as femtoseconds under conditions that maximize the extent of reaction initiation throughout the crystal.

scholarly journals Viscous medium-based crystal support in sample holder for fixed-target serial femtosecond crystallography

2020 ◽  
Author(s):  
Keondo Lee ◽  
Donghyeon Lee ◽  
Sangwon Baek ◽  
Jaehyun Park ◽  
Sang Jae Lee ◽  
...  
Keyword(s):  
Room Temperature ◽  
Polyimide Film ◽  
Glucose Isomerase ◽  
Viscous Medium ◽  
Sample Holder ◽  
Polyimide Films ◽  
Fixed Target ◽  
Time Resolved ◽  
Viscous Media ◽  
Serial Femtosecond Crystallography

AbstractSerial femtosecond crystallography (SFX) enables the determination of a room-temperature crystal structure of macromolecules without causing radiation damage, as well as provides time-resolved molecular dynamics data in pump-probe experiments. Fixed-target SFX (FT-SFX) can minimize sample consumption and physical effects to crystals during sample delivery. Various types of sample holders have been developed and applied in FT-SFX; however, no sample holder has been developed that can universally mount crystals of various sizes and shapes. Here, we introduce a viscous media-based crystal support in a sample holder for FT-SFX. Crystal samples were embedded in viscous media such as gelatin and agarose, which were enclosed in a polyimide film. In the vertically placed sample holder, the viscous medium stably supported crystals between the two polyimide films without crystal sinking due to gravity. Using this method, we performed FT-SFX experiments with glucose isomerase and lysozyme embedded in gelatin and agarose, respectively. The room-temperature crystal structures of glucose isomerase and lysozyme were successfully determined at 1.75 and 1.80 Å resolutions, respectively. Viscous media used in this experiment showed negligible background scattering in data processing. This method is useful for delivering crystal samples of various sizes and shapes in FT-SFX experiments.

scholarly journals Advances in Diffraction Studies of Light-Induced Transient Species in Molecular Crystals and Selected Complementary Techniques

Crystals ◽  
2021 ◽  
Vol 11 (11) ◽  
pp. 1345
Author(s):  
Krystyna A. Deresz ◽  
Piotr Łaski ◽  
Radosław Kamiński ◽  
Katarzyna N. Jarzembska
Keyword(s):  
Free Electron Lasers ◽  
Transient Species ◽  
X Ray Diffraction ◽  
Time Resolved ◽  
X Ray ◽  
Recent Developments ◽  
Complementary Techniques ◽  
X Ray Absorption ◽  
Serial Femtosecond Crystallography ◽  
Major Emphasis

The review provides a summary of the current methods of tracing photoexcitation processes and structural dynamics in the solid state, putting major emphasis on the X-ray diffraction techniques (time-resolved Laue diffraction on synchrotron sources and time-resolved serial femtosecond crystallography on X-ray free-electron lasers). The recent developments and nowadays experimental possibilities in the field are discussed along with the data processing and analysis approaches, and illustrated with some striking literature examples of the respective successful studies. Selected complementary methods, such as ultrafast electron diffraction or time-resolved X-ray absorption spectroscopy, are briefly presented.

Sign in / Sign up

Export Citation Format

Share Document