A5085398715- Photoreceptor and optogenetics research
- Enzyme Structure and Function
- Photosynthetic Processes and Mechanisms
- Photochromic and Fluorescence Chemistry
- Protein Structure and Dynamics
- Mass Spectrometry Techniques and Applications
- Receptor Mechanisms and Signaling
- Light effects on plants
- Advanced Electron Microscopy Techniques and Applications
- Advanced Fluorescence Microscopy Techniques
- Glycosylation and Glycoproteins Research
- Biochemical and Molecular Research
- X-ray Spectroscopy and Fluorescence Analysis
- Neuroscience and Neuropharmacology Research
- Neural dynamics and brain function
- Retinal Development and Disorders
- Spectroscopy and Quantum Chemical Studies
- Nuclear Physics and Applications
- Microtubule and mitosis dynamics
- Angiogenesis and VEGF in Cancer
- Molecular Communication and Nanonetworks
- Advanced Proteomics Techniques and Applications
- Advanced X-ray Imaging Techniques
- Metabolism and Genetic Disorders
- Crystallography and molecular interactions
Paul Scherrer Institute
2016-2025
Goethe Institute
2024
Goethe University Frankfurt
2024
MAX IV Laboratory
2024
University of Vienna
2020
Swiss Light Source
2014-2018
Max Planck Institute of Biophysics
2011-2015
University of Toronto
2015
Structural Genomics Consortium
2015
Max Planck Society
2004
Look fast Organisms from bacteria to humans sense and react light. Proteins that contain the light-sensitive molecule retinal couple absorption of light conformational changes produce a signal or move ions across membrane. Nogly et al. used an x-ray laser probe earliest structural chromophore within microcrystals ion pump bacteriorhodopsin (see Perspective by Moffat). The excited-state wiggles but is held in place so only one double bond capable isomerizing. A water adjacent proton-pumping...
Abstract Historically, room-temperature structure determination was succeeded by cryo-crystallography to mitigate radiation damage. Here, we demonstrate that serial millisecond crystallography at a synchrotron beamline equipped with high-viscosity injector and high frame-rate detector allows typical crystallographic experiments be performed room-temperature. Using crystal scanning approach, determine the high-resolution of sensitive molybdenum storage protein, soaking drug colchicine into...
Refilling the proton pump Proteins are dynamic. Rearrangements of side chains, secondary structure, and entire domains gate functional transitions on time scales ranging from picoseconds to milliseconds. Weinert et al. used time-resolved serial crystallography study large conformational changes in bacteriorhodopsin that allow for redistribution protons during pumping cycle. They adapted methods x-ray free electron lasers synchrotron sources. Large loop movements a chain water molecules were...
Abstract Vision is initiated by the rhodopsin family of light-sensitive G protein-coupled receptors (GPCRs) 1 . A photon absorbed 11- cis retinal chromophore rhodopsin, which isomerizes within 200 femtoseconds to all- trans conformation 2 , thereby initiating cellular signal transduction processes that ultimately lead vision. However, intramolecular mechanism photoactivated induces activation events inside remains experimentally unclear. Here we use ultrafast time-resolved crystallography at...
Chloride transport by microbial rhodopsins is an essential process for which molecular details such as the mechanisms that convert light energy to drive ion pumping and ensure unidirectionality of have remained elusive. We combined time-resolved serial crystallography with spectroscopy multiscale simulations elucidate mechanism a chloride-pumping rhodopsin structural dynamics throughout cycle. traced transient anion-binding sites, obtained evidence how used in mechanism, identified steric...
Photolyases, a ubiquitous class of flavoproteins, use blue light to repair DNA photolesions. In this work, we determined the structural mechanism photolyase-catalyzed cyclobutane pyrimidine dimer (CPD) lesion using time-resolved serial femtosecond crystallography (TR-SFX). We obtained 18 snapshots that show time-dependent changes in four reaction loci. used these results create movie depicts CPD lesions picosecond-to-nanosecond range, followed by recovery enzymatic moieties involved...
The CC chemokine receptor 7 (CCR7) balances immunity and tolerance by homeostatic trafficking of immune cells. In cancer, CCR7-mediated leads to lymph node metastasis, suggesting the as a promising therapeutic target. Here, we present crystal structure human CCR7 fused protein Sialidase NanA using data up 2.1 Å resolution. shows ligand Cmp2105 bound an intracellular allosteric binding pocket. A sulfonamide group, characteristic for various ligands, binds patch conserved residues in Gi region...
The dynamic instability of microtubules plays a key role in controlling their organization and function, but the cellular mechanisms regulating this process are poorly understood. Here, we show that cytoplasmic linker-associated proteins (CLASPs) suppress transitions from microtubule growth to shortening, termed catastrophes, including those induced by microtubule-destabilizing agents physical barriers. Mammalian CLASPs encompass three TOG-like domains, TOG1, TOG2, TOG3, none which bind free...
Here, a method for presenting crystals of soluble and membrane proteins growing in the lipid cubic or sponge phase situ diffraction data collection at cryogenic temperatures is introduced. The dispenses with need technically demanding inefficient crystal-harvesting step that an integral part meso crystals. Crystals are dispersed bolus mesophase sandwiched between thin plastic windows. contains tens to hundreds crystals, visible in-line microscope macromolecular crystallography synchrotron...
Photoswitchable reagents are powerful tools for high-precision studies in cell biology. When these globally administered yet locally photoactivated two-dimensional (2D) cultures, they can exert micron- and millisecond-scale biological control. This gives them great potential use biologically more relevant three-dimensional (3D) models
The binding and release of ligands from their protein targets is central to fundamental biological processes as well drug discovery. Photopharmacology introduces chemical triggers that allow the changing ligand affinities thus activity by light. Insight into molecular mechanisms photopharmacology largely missing because relevant transitions during light-triggered reaction cannot be resolved conventional structural biology. Using time-resolved serial crystallography at a synchrotron X-ray...
Selective coupling of G protein (heterotrimeric guanine nucleotide-binding protein)-coupled receptors (GPCRs) to specific Gα-protein subtypes is critical transform extracellular signals, carried by natural ligands and clinical drugs, into cellular responses. At the center this transduction event lies formation a signaling complex between receptor protein. We report crystal structure light-sensitive GPCR rhodopsin bound an engineered mini-Go The conformation identical all previous structures...
Macromolecular crystallography has historically provided the atomic structures of proteins fundamental to cellular functions. However, advent cryo-electron microscopy for structure determination large and increasingly smaller flexible signaled a paradigm shift in structural biology. The extensive sequence data from advanced sequencing techniques have been pivotal training computational models accurate prediction, unveiling general fold most proteins. Here, we present perspective on rise...
Abstract Charge-transfer reactions in proteins are important for life, such as photolyases which repair DNA, but the role of structural dynamics remains unclear. Here, using femtosecond X-ray crystallography, we report changes that take place while electrons transfer along a chain four conserved tryptophans Drosophila melanogaster (6-4) photolyase. At femto- and picosecond delays, photoreduction flavin by first tryptophan causes directed responses at key asparagine, salt bridge,...
The Parallel Robotics Inspired Goniometer (PRIGo) is a novel compact and high-precision goniometer providing an alternative to (mini-)kappa, traditional three-circle goniometers Eulerian cradles used for sample reorientation in macromolecular crystallography. Based on combination of serial parallel kinematics, PRIGo emulates arc. It mounted air-bearing stage rotation around ω consists four linear positioners working synchronously achieve x, y, z translations χ (0-90°), followed by ϕ (0-360°)...
Recent improvements in data-collection strategies have pushed the limits of native SAD (single-wavelength anomalous diffraction) phasing, a method that uses weak signal light elements naturally present macromolecules. These involve merging multiple data sets from either crystals or single crystal collected orientations at low X-ray dose. Both approaches yield high multiplicity while minimizing radiation damage and systematic error, thus ensuring accurate measurements differences. Here,...
Significance Adenylyl and guanylyl cyclases are at the core of cellular signaling. Although molecular mechanisms reactions catalyzed by these enzymes well established, their structures biophysical properties remain only partially characterized. Here, we report structure cytosolic domain a mycobacterial adenylyl cyclase Cya, an evolutionary ancestor mammalian membrane cyclases. The reveals helical domain, highly conserved structural element that links catalytic transmembrane portions Cya. We...
DM9 domain was first identified in Drosophila melanogaster, and it subsequently found to integrate with or without other protein domains across a wide range of invertebrates vertebrates. In the present study, member containing (DM9CP) family from marine invertebrate Crassostrea gigas (designated CgDM9CP-1), which only composed two domains, taken as model study biological functions its molecular determinants. CgDM9CP-1 exhibit high binding specificity avidity toward d-mannose residue. It...
Serial and time-resolved macromolecular crystallography are on the rise. However, beam time at X-ray free-electron lasers is limited most third-generation synchrotron-based beamlines do not offer necessary infrastructure yet. Here, a new setup demonstrated, based JUNGFRAU detector Jungfraujoch data-acquisition system, that enables collection of kilohertz serial data fourth-generation synchrotrons. More importantly, it shown this capable collecting multiple-time-point protein dynamics rates,...