A5080468830- Enzyme Structure and Function
- Antibiotic Resistance in Bacteria
- Protein Structure and Dynamics
- Erythrocyte Function and Pathophysiology
- X-ray Diffraction in Crystallography
- Biochemical and Molecular Research
- Peptidase Inhibition and Analysis
- Crystallization and Solubility Studies
- Advanced X-ray Imaging Techniques
- Bacteriophages and microbial interactions
- Experimental and Theoretical Physics Studies
- Bacterial Genetics and Biotechnology
- Probiotics and Fermented Foods
- HIV/AIDS drug development and treatment
University Medical Center Hamburg-Eppendorf
2022-2024
Universität Hamburg
2022-2024
Serial crystallography at conventional synchrotron light sources (SSX) offers the possibility to routinely collect data room temperature using micrometre-sized crystals of biological macromolecules. However, SSX collection is not yet as routine and currently takes significantly longer than standard rotation series cryo-crystallography. Thus, its use for high-throughput approaches, such fragment-based drug screening, where measure physiological temperatures would be a great benefit, impaired....
Abstract We introduce the spitrobot , a protein crystal plunger, enabling reaction quenching via cryo-trapping with time-resolution in millisecond range. Protein crystals are mounted on canonical micromeshes an electropneumatic piston, where kept humidity and temperature-controlled environment, then reactions initiated liquid application method (LAMA) plunging into nitrogen is after electronically set delay time to cryo-trap intermediate states. High-magnification images automatically...
Abstract β-lactamases are a major cause of rapidly emerging and spreading antibiotic resistance. Currently β-lactamase inhibitors (BLIs) in clinical use act only on Ambler Class A, C some class D lactamases. The urgent need to identify new BLIs recently lead FDA approval boron-based compounds BLIs, e.g. Vaborbactam. proteasome Bortezomib Ixazomib used cancer therapy as multiple myeloma drugs but they also bind Ser-/Thr- proteases. In this study we show the crystal structures CTX-M-14 with...
Fixed targets are a versatile sample delivery method for serial crystallography experiments and favorable choice regarding efficiency. The hit-and-return (HARE) chips utilize pyramidal-shaped features to pre-position crystals in regular pattern. Typically, these micron-sized chemically etched into single-crystal silicon wafers providing rigid mount with minimal background. As polymer emerging as an alternative, here we report the production of HARE identical micron accuracy by laser ablating...
Abstract The emergence and spread of antibiotic resistance represent a growing threat to public health. Of particular concern is the appearance β-lactamases, which are capable hydrolyze inactivate most important class antibiotics, β-lactams. Effective β-lactamase inhibitors mechanistic insights into their action central in overcoming this type resistance, context boronate-based were just recently approved treat multidrug-resistant bacteria. Using boric acid as simplified inhibitor model,...
Abstract We present an environmental enclosure for fixed-target serial crystallography, enabling X-ray diffraction experiments in a temperature window from below 10 °C to above 70 - universal parameter of protein function. Via 5D-SSX time-resolved can now be carried out at physiological temperatures, providing fundamentally new insights into show temperature-dependent modulation turnover kinetics the mesophilic β -lactamase CTX-M-14 and hyperthermophilic enzyme xylose isomerase.
<title>Abstract</title> β-lactamase inhibitors are central in overcoming emerging antibiotic resistance, and boronate-based were just recently approved to treat multidrug-resistant bacteria. Using boric acid as a simplified inhibitor model, time-resolved serial crystallography was employed obtain mechanistic insights into binding the active site serine of CTX M 14, identifying reaction time frame 80 – 100 ms. In next step, subsequent 1,2-diol ester formation with glycerol monitored...
Abstract We introduce the spitrobot , a protein crystal plunger, enabling reaction quenching via cryo-trapping with millisecond time-resolution. Canonical micromesh loops are mounted on an electropneumatic piston, reactions initiated liquid application method (LAMA), and finally intermediate states cryo-trapped in nitrogen. demonstrate binding of several ligands microcrystals three enzymes, trapping intermediates conformational changes macroscopic crystals tryptophan synthase.