Katerina Dörner
A5087717325- Enzyme Structure and Function
- Advanced Electron Microscopy Techniques and Applications
- Advanced X-ray Imaging Techniques
- Photosynthetic Processes and Mechanisms
- Metalloenzymes and iron-sulfur proteins
- X-ray Spectroscopy and Fluorescence Analysis
- Mitochondrial Function and Pathology
- Protein Structure and Dynamics
- Light effects on plants
- bioluminescence and chemiluminescence research
- HIV Research and Treatment
- Photoreceptor and optogenetics research
- Monoclonal and Polyclonal Antibodies Research
- Microbial Fuel Cells and Bioremediation
- Metabolism and Genetic Disorders
- Bacteriophages and microbial interactions
- Advanced Radiotherapy Techniques
- Biochemical and Molecular Research
- CO2 Reduction Techniques and Catalysts
- Electron Spin Resonance Studies
- Porphyrin Metabolism and Disorders
- Advanced Fluorescence Microscopy Techniques
- Particle Accelerators and Free-Electron Lasers
- Metal-Catalyzed Oxygenation Mechanisms
- Molecular spectroscopy and chirality
European X-Ray Free-Electron Laser
2018-2025
Arizona State University
2014-2018
Universität Hamburg
2015-2018
Center for Free-Electron Laser Science
2016-2018
University of Freiburg
1917-2017
Deutsches Elektronen-Synchrotron DESY
2015-2017
University of Minnesota
2017
Max Planck Institute for the Structure and Dynamics of Matter
2017
Delft University of Technology
2012
Allgemeines Krankenhaus Celle
2009
Abstract The new European X-ray Free-Electron Laser is the first free-electron laser capable of delivering pulses with a megahertz inter-pulse spacing, more than four orders magnitude higher previously possible. However, to date, it has been unclear whether would indeed be possible measure high-quality diffraction data at pulse repetition rates. Here, we show that structures can obtained using currently available operating conditions XFEL. We present two complete sets, one from well-known...
Abstract Serial femtosecond crystallography requires reliable and efficient delivery of fresh crystals across the beam an X-ray free-electron laser over course experiment. We introduce a double-flow focusing nozzle to meet this challenge, with significantly reduced sample consumption, while improving jet stability previous generations nozzles. demonstrate its use determine first room-temperature structure RNA polymerase II at high resolution, revealing new structural details. Moreover,...
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 crystallography is one of the main methods to determine atomic-resolution 3D images whole spectrum molecules ranging from small inorganic clusters large protein complexes consisting hundred-thousands atoms that constitute macromolecular machinery life. Life not static, and unravelling structure dynamics most important reactions in chemistry biology essential uncover their mechanism. Many these reactions, including photosynthesis which drives our biosphere, are light induced occur on...
Abstract The emergence of high repetition-rate X-ray free-electron lasers (XFELs) powered by superconducting accelerator technology enables the measurement significantly more experimental data per day than was previously possible. European XFEL is expected to provide 27,000 pulses second, over two orders magnitude any other XFEL. increased pulse rate a key enabling factor for single-particle diffractive imaging, which relies on averaging weak diffraction signal from single biological...
The Sample Environment and Characterization (SEC) group of the European X-ray Free-Electron Laser (EuXFEL) develops sample delivery systems for various scientific instruments, including injection liquid samples that enable serial femtosecond crystallography (SFX) single-particle imaging (SPI) experiments, among others. For rapid prototyping device types materials, sub-micrometre precision 3D printers are used to address specific experimental conditions SFX SPI by providing a large number...
Aromatic compounds are widely distributed in nature and can only be biomineralized by microorganisms. In anaerobic bacteria, benzoyl-CoA (BCoA) is a central intermediate of aromatic degradation, serves as substrate for dearomatizing BCoA reductases (BCRs). facultative anaerobes, the mechanistically difficult reduction to cyclohexa-1,5-dienoyl-1-carboxyl-CoA (dienoyl-CoA) driven stoichiometric ATP hydrolysis, catalyzed soluble, three [4Fe-4S] cluster-containing BCR. this work, an vitro assay...
Abstract The world’s first superconducting megahertz repetition rate hard X-ray free-electron laser (XFEL), the European XFEL, began operation in 2017, featuring a unique pulse train structure with 886 ns between pulses. With its rapid rate, XFEL may alleviate some of increasing demand for beamtime, particularly membrane protein serial femtosecond crystallography (SFX), leveraging orders-of-magnitude faster data collection. Here, we report SFX experiment, where determined 2.9 Å-resolution...
The understanding of signal transduction mechanisms in photoreceptor proteins is essential for elucidating how living organisms respond to light as environmental stimuli. In this study, we investigated the ATP binding, photoactivation and process photoactivatable adenylate cyclase from Oscillatoria acuminata (OaPAC) upon blue excitation. Structural models with bound active site native OaPAC at cryogenic well room temperature are presented. found one conformation cryogenic- two conformations...
Abstract Respiratory complex I converts the free energy of ubiquinone reduction by NADH into a proton motive force, redox reaction catalyzed flavin mononucleotide(FMN) and chain seven iron–sulfur centers. Electron transfer rates between centers were determined ultrafast freeze‐quenching analysis EPR UV/Vis spectroscopy. The rapidly oxidizes three molecules. electron‐tunneling rate most distant in middle depends on state center N2 at end chain, is sixfold slower when reduced. conformational...
The NADH:ubiquinone oxidoreductase (complex I) from Escherichia coli is composed of 13 subunits called NuoA through NuoN. It catalyzes the electron transfer NADH to ubiquinone by a chain redox groups consisting one FMN and seven iron-sulfur clusters. function additional, nonconserved cluster N7 located on NuoG not known. has been speculated that it involved in transfer, due its distance more than 20 A chain. Dithionite-reduced minus NADH-reduced EPR difference spectra complex I soluble...
The NADH:ubiquinone oxidoreductase, respiratory complex I, couples the transfer of electrons from NADH to ubiquinone with a translocation protons across membrane. consists peripheral arm catalyzing electron reaction and membrane involved in proton translocation. recently published X-ray structures revealed presence unique 110 Å "horizontal" helix aligning arm. On basis this finding, it was proposed that energy released by redox is transmitted via conformational change horizontal helix....
Here we present a new approach to diffraction imaging of amyloid fibrils, combining free-standing graphene support and single nanofocused X-ray pulses femtosecond duration from an free-electron laser. Due the very low background scattering mutual alignment filaments, tobacco mosaic virus (TMV) filaments protofibrils is obtained 2.7 Å 2.4 resolution in patterns, respectively. Some TMV patterns exhibit asymmetry that indicates presence limited number axial rotations XFEL focus. Signal-to-noise...
Serial femtosecond crystallography (SFX) with X-ray free electron lasers (XFELs) allows structure determination of membrane proteins and time-resolved crystallography. Common liquid sample delivery continuously jets the protein crystal suspension into path XFEL, wasting a vast amount due to pulsed nature all current XFEL sources. The European (EuXFEL) delivers (fs) pulses in trains spaced 100 ms apart whereas within are currently separated by 889 ns. Therefore, continuous via fast wastes...
The SPB/SFX instrument of the European XFEL provides unique possibilities for high-throughput serial femtosecond crystallography. This publication presents liquid-jet sample delivery setup this instrument. is compatible with state-of-the-art gas dynamic virtual nozzle systems as well high-viscosity extruders and space flexibility other liquid injection devices future upgrades. jets are confined in a differentially pumped catcher assembly can be replaced within couple minutes through...
Free-electron lasers (FEL) are revolutionizing X-ray-based structural biology methods. While protein crystallography is already routinely performed at FELs, Small Angle X-ray Scattering (SAXS) studies of biological macromolecules not as prevalent. SAXS allows the study shape and overall structure proteins nucleic acids in solution, a quasi-native environment. In chemical biophysical parameters that have an influence on dynamics molecules can be varied their effect conformational changes...
Time-resolved crystallography enables the visualization of protein molecular motion during a reaction. Although light is often used to initiate reactions in time-resolved crystallography, only small number proteins can be activated by light. However, many biological triggered interaction between and ligands. The sample delivery method presented here uses mix-and-extrude approach based on 3D-printed microchannels conjunction with micronozzle. diffusive mixing study dynamics samples viscous...
The Lysinibacillus sphaericus proteins Tpp49Aa1 and Cry48Aa1 can together act as a toxin toward the mosquito Culex quinquefasciatus have potential use in biocontrol. Given that with sequence homology to individual activity alone against other insect species, structure of was solved order understand this protein more fully inform design improved biopesticides. is naturally expressed crystalline inclusion within host bacterium, MHz serial femtosecond crystallography using novel nanofocus...
Penicillin-binding protein 2a (PBP2a) is a transpeptidase responsible for the β-lactam resistance in methicillin-resistant Staphylococcus aureus (MRSA), posing significant challenges to antibiotic therapy. PBP2a's unique structural features, including its highly flexible active site and allosteric regulation, enable it maintain catalytic activity even presence of antibiotics. Despite extensive characterization using cryogenic crystallography, key questions remain about dynamic properties...
The advent and application of the X-ray free-electron laser (XFEL) has uncovered structures proteins that could not previously be solved using traditional crystallography. While this new technology is powerful, optimization process still needed to improve data quality analysis efficiency. One area sample heterogeneity, where variations in crystal size (among other factors) lead requirement large sets (and thus 10–100 mg protein) for determining accurate structure factors. To decrease...
Abstract The European X-ray Free Electron Laser (XFEL) and Linac Coherent Light Source (LCLS) II are extremely intense sources of X-rays capable generating Serial Femtosecond Crystallography (SFX) data at megahertz (MHz) repetition rates. Previous work has shown that it is possible to use consecutive pulses collect diffraction patterns from individual crystals. Here, we exploit the MHz pulse structure XFEL obtain two complete datasets same lysozyme crystal, first hit second hit, before exits...