A5028458095- Enzyme Structure and Function
- Advanced X-ray Imaging Techniques
- Advanced Electron Microscopy Techniques and Applications
- Photoreceptor and optogenetics research
- Viral Infectious Diseases and Gene Expression in Insects
- Photosynthetic Processes and Mechanisms
- Biofield Effects and Biophysics
- Advanced Fluorescence Microscopy Techniques
- X-ray Spectroscopy and Fluorescence Analysis
- Crystallography and Radiation Phenomena
- Chemical Reactions and Isotopes
- X-ray Diffraction in Crystallography
- Hemoglobin structure and function
- Spectroscopy and Quantum Chemical Studies
- Nanoparticle-Based Drug Delivery
- Laser-Plasma Interactions and Diagnostics
- Healthcare and Venom Research
- Atomic and Molecular Physics
- High-pressure geophysics and materials
- Fungal and yeast genetics research
- Molecular Communication and Nanonetworks
- Enzyme Catalysis and Immobilization
- Photochromic and Fluorescence Chemistry
- RNA and protein synthesis mechanisms
- Granular flow and fluidized beds
Max Planck Institute for Medical Research
2016-2024
Max Planck Society
2021
Abstract Bacteriorhodopsin (bR) is a light-driven proton pump. The primary photochemical event upon light absorption isomerization of the retinal chromophore. Here we used time-resolved crystallography at an X-ray free-electron laser to follow structural changes in multiphoton-excited bR from 250 femtoseconds 10 picoseconds. Quantum chemistry and ultrafast spectroscopy were identify sequential two-photon process, leading excitation tryptophan residue flanking chromophore, as first...
Light makes light work of fatty acids Photosynthetic organisms are notable for their ability to capture energy and use it power biosynthesis. Some algae have gone a step beyond photosynthesis can initiate enzymatic photodecarboxylation acids, producing long-chain hydrocarbons. To understand this transformation, Sorigué et al. brought bear an array structural, computational, spectroscopic techniques fully characterized the catalytic cycle enzyme. These experiments consistent with mechanism...
Abstract High-intensity femtosecond pulses from an X-ray free-electron laser enable pump–probe experiments for the investigation of electronic and nuclear changes during light-induced reactions. On timescales ranging femtoseconds to milliseconds a variety biological systems, time-resolved serial crystallography (TR-SFX) has provided detailed structural data isomerization, breakage or formation chemical bonds electron transfer 1,2 . However, all ultrafast TR-SFX studies date have employed...
Abstract X-ray free-electron lasers (XFELs) enable novel experiments because of their high peak brilliance and femtosecond pulse duration. However, non-superconducting XFELs offer repetition rates only 10–120 Hz, placing significant demands on beam time sample consumption. We describe serial crystallography performed at the European XFEL, first MHz rate delivering 1.128 trains 10 Hz. Given short spacing between pulses, damage caused by shock waves launched one XFEL probed subsequent pulses...
X-ray free-electron lasers (XFELs) enable crystallographic structure determination beyond the limitations imposed upon synchrotron measurements by radiation damage. The need for very short XFEL pulses is relieved through gating of Bragg diffraction loss crystalline order as damage progresses, but not if ionization events are spatially non-uniform due to underlying elemental distributions, in biological samples. Indeed, correlated movements iron and sulfur ions were observed XFEL-irradiated...
Serial femtosecond crystallography (SFX) at X-ray free-electron lasers (XFELs) offers unprecedented possibilities for macromolecular structure determination of systems that are prone to radiation damage. However, phasing XFEL data de novo is complicated by the inherent inaccuracy SFX data, and only a few successful examples, mostly based on exceedingly strong anomalous or isomorphous difference signals, have been reported. Here, it shown from thaumatin microcrystals can be successfully...
Crystallography chips are fixed-target supports consisting of a film (for example Kapton) or wafer silicon) that is processed using semiconductor-microfabrication techniques to yield an array wells through-holes in which single microcrystals can be lodged for raster-scan probing. Although relatively expensive fabricate, offer efficient means high-throughput sample presentation serial diffraction data collection at synchrotron X-ray free-electron laser (XFEL) sources. Truly loading chip (one...
Abstract X-ray free-electron lasers (XFELs) enable obtaining novel insights in structural biology. The recently available MHz repetition rate XFELs allow full data sets to be collected shorter time and can also decrease sample consumption. However, the microsecond spacing of XFEL pulses raises new challenges, including possible damage induced by shock waves that are launched preceding sample-carrying jet. We explored this matter with an X-ray-pump/X-ray-probe experiment employing haemoglobin...
Abstract Serial femtosecond crystallography at X-ray free-electron lasers (XFELs) offers unprecedented possibilities for macromolecular structure determination of systems prone to radiation damage. However, de novo determination, i.e., without prior structural knowledge, is complicated by the inherent inaccuracy serial data. By its very nature, data collection entails shot-to-shot fluctuations in wavelength and intensity as well variations crystal size quality that must be averaged out....
Abstract Reversibly photoswitchable fluorescent proteins are essential markers for advanced biological imaging, and optimization of their photophysical properties underlies improved performance novel applications. Here we establish a link between photoswitching contrast, one the key parameters that dictate achievable resolution in nanoscopy applications, chromophore conformation non‐fluorescent state rsEGFP2, widely employed label REversible Saturable OpticaL Fluorescence Transitions...
Serial crystallography (SX) efficiently distributes over many crystals the radiation dose absorbed during diffraction data acquisition, enabling structure determination of samples at ambient temperature. SX relies on rapid and reliable replacement X-ray-exposed with fresh a rate commensurate acquisition rate. `Solid supports', also known as `fixed targets' or `chips', offer one approach. These are microscopically thin solid panes into onto which deposited to be individually interrogated by...
Shock waves launched by x-ray pulses in sample-carrying liquid jets may affect protein crystallography data collected at MHz repetition rate free-electron laser (XFEL) facilities, damaging the crystals before they are probed. We investigated shock damage lysozyme microcrystals using a double-pulse operation mode low facility. The generated with pressures that covered and exceeded expected pulse experiments European XFEL (EuXFEL) laser. quality of diffraction from was degraded after passed. A...
Summary High-intensity femtosecond pulses from an X-ray free-electron laser enable pump probe experiments for investigating electronic and nuclear changes during light-induced reactions. On time scales ranging femtoseconds to milliseconds a variety of biological systems, time-resolved serial crystallography (TR-SFX) has provided detailed structural data isomerization, breakage or formation chemical bonds electron transfer 1 . However, all ultra-fast TR-SFX studies date have employed such...
Serial femtosecond crystallography (SFX) data were recorded at the European X-ray free-electron laser facility (EuXFEL) with protein microcrystals delivered via a microscopic liquid jet. An XFEL beam striking such jet may launch supersonic shock waves up jet, compromising oncoming sample. To investigate this efficiently, we employed novel pulse pattern to nominally expose sample between zero and four before being probed. Analyzing hit rate, indexing resolution for diffraction MHz rates,...
We provide a detailed description of serial femtosecond crystallography (SFX) dataset collected at the European X-ray free-electron laser facility (EuXFEL). The EuXFEL is first high repetition rate XFEL delivering MHz pulse trains 10 Hz. short spacing (<1 µs) between pulses requires fast flowing microjets for sample injection and frame detectors. A data set was recorded microcrystalline mixture least three different jack bean proteins (urease, concanavalin A, B). one megapixel Adaptive Gain...
Abstract We provide a detailed description of gadoteridol-derivatized lysozyme (gadolinium lysozyme) two-colour serial femtosecond crystallography (SFX) dataset for multiple wavelength anomalous dispersion (MAD) structure determination. The data was collected at the Spring-8 Angstrom Compact free-electron LAser (SACLA) facility using double-pulse beam to record two diffraction patterns simultaneously in one image. Gadolinium chosen as well-established model system that has very strong...
Abstract Reversibly photoswitchable fluorescent proteins are essential markers for advanced biological imaging, and optimization of their photophysical properties underlies improved performance novel applications. Here we establish a link between photoswitching contrast, key parameter that largely dictates the achievable resolution in nanoscopy applications, chromophore conformation non-fluorescent state rsEGFP2, widely employed label REversible Saturable OpticaL Fluorescence Transitions...
Using X-ray free electron laser (XFEL) radiation to conduct an pump probe experiment, we studied strongly ionized water as part of our ongoing work on damage. After irradiance with a pulse nominal fluence ~$5 \times 10^5$ J/cm$^2$, observed for pump-probe delays 75 fs and longer unexpected structural rearrangement, exhibiting characteristic length scale ~9 \r{A}. Simulations suggest that the experiment probes superposition in two distinct regimes. In first, fluences expected at focus create...
Abstract In the warm dense matter (WDM) regime, where condensed, gas, and plasma phases coexist, frequently exhibits unusual properties that cannot be described by contemporary theory. Experiments reporting phenomena in WDM are therefore of interest to advance our physical understanding this which is found dwarf stars, giant planets, fusion ignition experiments. Using 7.1 keV X-ray free electron laser radiation (nominally 5×10 5 J/cm 2 ), we produced probed transient liquid water. Wide-angle...