A5039204846- Photoreceptor and optogenetics research
- Photochromic and Fluorescence Chemistry
- Enzyme Structure and Function
- Photosynthetic Processes and Mechanisms
- Spectroscopy and Quantum Chemical Studies
- Light effects on plants
- Click Chemistry and Applications
- bioluminescence and chemiluminescence research
- Protein Degradation and Inhibitors
- Neural dynamics and brain function
- Biochemical and Molecular Research
- Neuroscience and Neuropharmacology Research
- Ubiquitin and proteasome pathways
- Nanoplatforms for cancer theranostics
- Computational Drug Discovery Methods
- Advanced Fluorescence Microscopy Techniques
- Advanced Electron Microscopy Techniques and Applications
- Solid-state spectroscopy and crystallography
- Porphyrin and Phthalocyanine Chemistry
- Glycosylation and Glycoproteins Research
- Carbohydrate Chemistry and Synthesis
- Photorefractive and Nonlinear Optics
- Electrochemical Analysis and Applications
- Receptor Mechanisms and Signaling
- Radical Photochemical Reactions
Paul Scherrer Institute
2019-2024
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...
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
Photolyase is an enzyme that uses light to catalyze DNA repair. To capture the reaction intermediates involved in enzyme's catalytic cycle, we conducted a time-resolved crystallography experiment. We found photolyase traps excited state of active cofactor, flavin adenine dinucleotide (FAD), highly bent geometry. This performs electron transfer damaged DNA, inducing show repair reaction, which involves lysis two covalent bonds, occurs through single-bond intermediate. The transformation...
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 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...
The structural response of the drosophila (6-4) photolyase to photoinduced electron transfer along a chain tryptophans is revealed using serial crystallographic snapshot protein in its radical pair state.
Here, we performed photo-SAR studies in a series of photoswitchable tubulin inhibiting antimitotics, and discovered that fluorescent antenna motifs permit their single-photon photoisomerisation biological conditions at up to >600 nm; were also able visualise the temporal onset long-term photopharmacological effects, using large-field-of-view microscopy. Previously, azobenzene analogues polymerisation inhibitor combretastatin A4 (PSTs) had been developed optically control microtubule...
Abstract Serial crystallography at X-ray free-electron lasers (XFELs) permits the determination of radiation-damage free static as well time-resolved protein structures room temperature. Efficient sample delivery is a key factor for such experiments. Here, we describe multi-reservoir, high viscosity extruder step towards automation XFELs. Compared to standard single extruder, exchange time was halved and workload users greatly reduced. In-built temperature control samples facilitated optimal...
Abstract Photoswitchable reagents to modulate microtubule stability and dynamics are an exciting tool approach towards micron- millisecond-scale control over endogenous cytoskeleton-dependent processes. When these globally administered yet locally photoactivated in 2D cell culture, they can exert precise biological that would have great potential for vivo translation across a variety of research fields all eukaryotes. However, photopharmacology’s reliance on the azobenzene photoswitch...
Serial data collection has emerged as a major tool for at state-of-the-art light sources, such microfocus beamlines synchrotrons and X-ray free-electron lasers. Challenging targets, characterized by small crystal sizes, weak diffraction stringent dose limits, benefit most from these methods. Here, the use of thin support made polymer-based membrane performing serial or screening experiments is demonstrated. It shown that supports are suitable wide range protein crystals suspended in liquids....
ABSTRACT Here we present GFP-orthogonal optically controlled reagents for reliable and repetitive in cellulo modulation of microtubule dynamics its dependent processes. Optically (“photopharmaceuticals”) have developed into powerful tools high-spatiotemporal-precision control endogenous biology, with numerous applications neuroscience, embryology, cytoskeleton research. However, the restricted chemical domain photopharmaceutical scaffolds has constrained their properties range applications....
G protein-coupled receptors (GPCRs) are the largest family of cell surface in humans. The binding and dissociation ligands tunes inherent conformational flexibility these important drug targets towards distinct functional states. Here we show how to trigger resolve protein-ligand interaction dynamics within human adenosine A
We develop reagents to optically control microtubule dynamics, with (1) lensless microscopy track their bioactivity; (2) photo-SAR find GFP-compatible reagents; & (3) fluorophore antennas drive single-photon isomerisation even ≥600 nm.
Abstract 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...
Abstract Serial crystallography at X-ray free-electron lasers (XFELs) permits the determination of radiation-damage free static as well time-resolved protein structures room temperature. Efficient sample delivery is a key factor for such experiments. Here, we describe multi-reservoir, high viscosity extruder step towards automation XFELs. Compared to standard single extruder, exchange time was halved and workload users greatly reduced. In-built temperature control samples facilitated optimal...