scholarly journals Double-focusing mixing jet for XFEL study of chemical kinetics

2014 ◽  
Vol 21 (6) ◽  
pp. 1364-1366 ◽  
Author(s):  
Dingjie Wang ◽  
Uwe Weierstall ◽  
Lois Pollack ◽  
John Spence
Keyword(s):  
Chemical Kinetics ◽  
Chemical Kinetic ◽  
Time Resolved ◽  
X Ray ◽  
Liquid Mixing ◽  
Combine Structure ◽  
Kinetic Mechanisms ◽  
Injection Methods ◽  
Double Focusing

Several liquid sample injection methods have been developed to satisfy the requirements for serial femtosecond X-ray nanocrystallography, which enables radiation-damage-free determination of molecular structure at room temperature. Time-resolved nanocrystallography would combine structure analysis with chemical kinetics by determining the structures of the transient states and chemical kinetic mechanisms simultaneously. A windowless liquid mixing jet device has been designed for this purpose. It achieves fast uniform mixing of substrates and enzymes in the jet within 250 µs, with an adjustable delay between mixing and probing by the X-ray free-electron laser beam of up to 1 s for each frame of a `movie'. The principle of the liquid mixing jet device is illustrated using numerical simulation, and experimental results are presented using a fluorescent dye.

scholarly journals Mix and Inject: Reaction Initiation by Diffusion for Time-Resolved Macromolecular Crystallography

2013 ◽  
Vol 2013 ◽  
pp. 1-10 ◽  
Author(s):  
Marius Schmidt
Keyword(s):  
Chemical Kinetics ◽  
Structure Determination ◽  
Macromolecular Crystallography ◽  
Time Resolved ◽  
X Ray ◽  
Transient States ◽  
Kinetic Mechanisms ◽  
Active Substrate ◽  
Reaction Initiation

Time-resolved macromolecular crystallography unifies structure determination with chemical kinetics, since the structures of transient states and chemical and kinetic mechanisms can be determined simultaneously from the same data. To start a reaction in an enzyme, typically, an initially inactive substrate present in the crystal is activated. This has particular disadvantages that are circumvented when active substrate is directly provided by diffusion. However, then it is prohibitive to use macroscopic crystals because diffusion times become too long. With small micro- and nanocrystals diffusion times are adequately short for most enzymes and the reaction can be swiftly initiated. We demonstrate here that a time-resolved crystallographic experiment becomes feasible by mixing substrate with enzyme nanocrystals which are subsequently injected into the X-ray beam of a pulsed X-ray source.

scholarly journals Time-Resolved Macromolecular Crystallography at Pulsed X-ray Sources

2019 ◽  
Vol 20 (6) ◽  
pp. 1401 ◽  
Author(s):  
Marius Schmidt
Keyword(s):  
Structural Biology ◽  
Free Electron ◽  
Reaction Dynamics ◽  
X Rays ◽  
Free Electron Lasers ◽  
Macromolecular Crystallography ◽  
Time Resolved ◽  
X Ray ◽  
Methodological Aspects

The focus of structural biology is shifting from the determination of static structures to the investigation of dynamical aspects of macromolecular function. With time-resolved macromolecular crystallography (TRX), intermediates that form and decay during the macromolecular reaction can be investigated, as well as their reaction dynamics. Time-resolved crystallographic methods were initially developed at synchrotrons. However, about a decade ago, extremely brilliant, femtosecond-pulsed X-ray sources, the free electron lasers for hard X-rays, became available to a wider community. TRX is now possible with femtosecond temporal resolution. This review provides an overview of methodological aspects of TRX, and at the same time, aims to outline the frontiers of this method at modern pulsed X-ray sources.

scholarly journals Determination of Excited State Molecular Structures from Time-Resolved Gas-Phase X-Ray Scattering

2020 ◽  
Author(s):  
Haiwang Yong ◽  
Andrés Moreno Carrascosa ◽  
Lingyu Ma ◽  
Brian Stankus ◽  
Michael P Minitti ◽  
...  
Keyword(s):  
Excited State ◽  
Gas Phase ◽  
Electronic States ◽  
Molecular Structures ◽  
Excited Electronic States ◽  
Comprehensive Investigation ◽  
Time Resolved ◽  
X Ray ◽  
X Ray Scattering

We present a comprehensive investigation of a recently introduced method to determine transient structures of molecules in excited electronic states with sub-Ångstrom resolution from time-resolved gas-phase scattering signals. The method,...

scholarly journals X-ray diagnostics of dense plasmas produced by high-energy heavy ions

1995 ◽  
Vol 13 (2) ◽  
pp. 183-190 ◽  
Author(s):  
W. Laux ◽  
J. Jacoby ◽  
D.H.H. Hoffmann
Keyword(s):  
Heavy Ions ◽  
High Energy ◽  
State Density ◽  
Time Resolved ◽  
X Ray ◽  
Dense Plasmas ◽  
Space And Time ◽  
Intensity Measurements ◽  
Near Future

Plasmas with solid-state density at temperatures of a few eV will be produced in the near future at GSI by high-energy heavy ions. To diagnose these plasmas, novel methods–based on hard X-ray diagnostics–have to be applied. An investigation of the hydrodynamic expansion of the target requires a space- and time-resolved measurement of the plasma density. The determination of the target density independently of its temperature is obtained by a special X-ray pinhole camera enabling a selective registration of the target K-shell emission in combination with space- and time-resolved beam intensity measurements.

scholarly journals Towards the time resolved X-ray structure determination of proteins

2012 ◽  
Vol 1817 ◽  
pp. S29-S30
Author(s):  
Raimund Fromme ◽  
Ingo Grotjohann ◽  
Christopher Kupitz ◽  
Shibom Basu ◽  
Petra Fromme
Keyword(s):  
Structure Determination ◽  
Time Resolved ◽  
X Ray

First performance assessment of the small-angle X-ray scattering beamline at ELETTRA

1998 ◽  
Vol 5 (3) ◽  
pp. 506-508 ◽  
Author(s):  
H. Amenitsch ◽  
M. Rappolt ◽  
M. Kriechbaum ◽  
H. Mio ◽  
P. Laggner ◽  
...  
Keyword(s):  
Small Angle ◽  
Wide Angle ◽  
Fibrous Materials ◽  
Time Resolved ◽  
X Ray ◽  
X Ray Scattering ◽  
Angle Scattering ◽  
Scattering Measurements ◽  
Double Focusing ◽  
Ray Scattering

The double-focusing high-flux wiggler beamline dedicated to small-angle X-ray scattering (SAXS) and wide-angle X-ray scattering (WAXS) at ELETTRA has gone into user operation recently. It has been designed specifically for time-resolved studies of non-crystalline and fibrous materials in the submillisecond time scale, and has been optimized for small-angle scattering measurements. An overview of the beamline status and of some representative results, highlighting the performance of the SAXS beamline, are given.

scholarly journals 3D-MiXD: 3D-printed X-ray-compatible microfluidic devices for rapid, low-consumption serial synchrotron crystallography data collection in flow

IUCrJ ◽  
2020 ◽  
Vol 7 (2) ◽  
pp. 207-219 ◽  
Author(s):  
Diana C. F. Monteiro ◽  
David von Stetten ◽  
Claudia Stohrer ◽  
Marta Sans ◽  
Arwen R. Pearson ◽  
...  
Keyword(s):  
Data Collection ◽  
Room Temperature ◽  
Time Resolved ◽  
X Ray ◽  
Biologically Relevant ◽  
3D Printed ◽  
Serial Crystallography ◽  
Low X ◽  
Very High

Serial crystallography has enabled the study of complex biological questions through the determination of biomolecular structures at room temperature using low X-ray doses. Furthermore, it has enabled the study of protein dynamics by the capture of atomically resolved and time-resolved molecular movies. However, the study of many biologically relevant targets is still severely hindered by high sample consumption and lengthy data-collection times. By combining serial synchrotron crystallography (SSX) with 3D printing, a new experimental platform has been created that tackles these challenges. An affordable 3D-printed, X-ray-compatible microfluidic device (3D-MiXD) is reported that allows data to be collected from protein microcrystals in a 3D flow with very high hit and indexing rates, while keeping the sample consumption low. The miniaturized 3D-MiXD can be rapidly installed into virtually any synchrotron beamline with only minimal adjustments. This efficient collection scheme in combination with its mixing geometry paves the way for recording molecular movies at synchrotrons by mixing-triggered millisecond time-resolved SSX.

scholarly journals Observation of substrate diffusion and ligand binding in enzyme crystals using high-repetition-rate mix-and-inject serial crystallography

IUCrJ ◽  
2021 ◽  
Vol 8 (6) ◽  
Author(s):  
Suraj Pandey ◽  
George Calvey ◽  
Andrea M. Katz ◽  
Tek Narsingh Malla ◽  
Faisal H. M. Koua ◽  
...  
Keyword(s):  
Repetition Rate ◽  
Initial Phase ◽  
High Repetition Rate ◽  
Irreversible Inhibitor ◽  
Time Resolved ◽  
X Ray ◽  
High Repetition ◽  
Substrate Diffusion ◽  
Crystal Volume

Here, we illustrate what happens inside the catalytic cleft of an enzyme when substrate or ligand binds on single-millisecond timescales. The initial phase of the enzymatic cycle is observed with near-atomic resolution using the most advanced X-ray source currently available: the European XFEL (EuXFEL). The high repetition rate of the EuXFEL combined with our mix-and-inject technology enables the initial phase of ceftriaxone binding to the Mycobacterium tuberculosis β-lactamase to be followed using time-resolved crystallography in real time. It is shown how a diffusion coefficient in enzyme crystals can be derived directly from the X-ray data, enabling the determination of ligand and enzyme–ligand concentrations at any position in the crystal volume as a function of time. In addition, the structure of the irreversible inhibitor sulbactam bound to the enzyme at a 66 ms time delay after mixing is described. This demonstrates that the EuXFEL can be used as an important tool for biomedically relevant research.

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