A5086090521- Photoreceptor and optogenetics research
- Enzyme Structure and Function
- Photosynthetic Processes and Mechanisms
- Metalloenzymes and iron-sulfur proteins
- Mass Spectrometry Techniques and Applications
- Antibiotic Resistance in Bacteria
- Receptor Mechanisms and Signaling
- Retinal Development and Disorders
- Neuroscience and Neuropharmacology Research
- Neural dynamics and brain function
- Polyoxometalates: Synthesis and Applications
- bioluminescence and chemiluminescence research
- Vitamin C and Antioxidants Research
- Tuberculosis Research and Epidemiology
- Photochromic and Fluorescence Chemistry
- Bioactive Compounds and Antitumor Agents
- ATP Synthase and ATPases Research
- Chemokine receptors and signaling
- Nicotinic Acetylcholine Receptors Study
- Microbial Metabolic Engineering and Bioproduction
- Nanocluster Synthesis and Applications
- Mycobacterium research and diagnosis
- Ocular and Laser Science Research
- Protein Structure and Dynamics
- Microbial Natural Products and Biosynthesis
Paul Scherrer Institute
2018-2024
Leiden University
2023-2024
Max Planck Institute of Biophysics
2016-2023
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...
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...
Abstract 8‐demethyl‐8‐aminoriboflavin‐5′‐phosphate (AFP) synthase (RosB) catalyzes the key reaction of roseoflavin biosynthesis by forming AFP from riboflavin‐5′‐phosphate (RP) and glutamate via intermediates 8‐demethyl‐8‐formylriboflavin‐5′‐phosphate (OHC‐RP) 8‐demethyl‐8‐carboxylriboflavin‐5′‐phosphate (HO 2 C‐RP). To understand this in which a methyl substituent an aromatic ring is replaced amine we structurally characterized RosB complex with OHC‐RP (2.0 Å) (1.7 Å). composed four...
Conserved residues are often considered essential for function, and substitutions in such expected to have a negative influence on the properties of protein. However, mutations few highly conserved β-lactamase from Mycobacterium tuberculosis, BlaC, were shown no or only limited effect enzyme. One mutant, D179N, even conveyed increased ceftazidime resistance upon bacterial cells, while displaying good activity against penicillins. The crystal structures BlaC D179N resting state complex with...
Cytochrome bd from Mycobacterium tuberculosis (Mtbd) is a menaquinol oxidase that has gained interest as an antibiotic target due to its importance in survival under infectious conditions. Mtbd contains characteristic disulfide bond been hypothesized allow for activity regulation at the enzymatic level, possibly helping M. rapidly adapt hostile environment of phagosome. Here, role and quinone specificity have determined by reconstitution minimal respiratory chain single-particle cryo-EM...
A continuous FeMo cofactor supply for nitrogenase maturation is ensured in Azotobacter vinelandii by developing a cage-like molybdenum storage protein (MoSto) capable to store ca. 120 molybdate molecules ( MoO42- ) as discrete polyoxometalate (POM) clusters. To gain mechanistic insight into this process, MoSto was characterized Mo and ATP/ADP content, structural, kinetic analysis. We defined three functionally relevant states specified the presence of both POM clusters (MoStofunct ), only...
The molybdenum storage protein (MoSto) deposits large amounts of as polyoxomolybdate clusters in a heterohexameric (αβ)3 cage-like complex under ATP consumption. Here, we suggest unique mechanism for the ATP-powered molybdate pumping process based on X-ray crystallography, cryoelectron microscopy, hydrogen-deuterium exchange mass spectrometry, and mutational studies MoSto from Azotobacter vinelandii. First, show that molybdate, ATP, Mg2+ consecutively bind into open ATP-binding groove...
Abstract Vision is initiated by the rhodopsin family of light-sensitive G protein-coupled receptors (GPCRs). A photon absorbed 11- cis retinal chromophore which isomerises within 200 femtoseconds to all- trans conformation, thereby initiating cellular signal transduction processes that ultimately lead vision. However, intramolecular mechanism photoactivated induces activation events inside remains elusive. In this work, we use ultrafast time-resolved crystallography at room temperature...
Serine β-lactamases inactivate β-lactam antibiotics in a two-step mechanism comprising acylation and deacylation. For the deacylation step, water molecule is activated by conserved glutamate residue to release adduct from enzyme. The third-generation cephalosporin ceftazidime poor substrate for class A β-lactamase BlaC Mycobacterium tuberculosis but it can be hydrolyzed faster when active site pocket enlarged, as was reported mutant P167S. conformational change Ω-loop of P167S displaces...
Abstract Channelrhodopsins, light-gated cation channels, enable precise control of neural cell depolarization or hyperpolarization with light in the field optogenetics. This study integrates time-resolved serial crystallography and atomistic molecular dynamics (MD) simulations to resolve structural changes during C1C2 channelrhodopsin activation. Our observations reveal that within crystal environment, predominantly remains a light-activated state characteristics M 390 intermediate. Here,...
Abstract Time-resolved serial crystallography at X-ray Free Electron Lasers offers the opportunity to observe ultrafast photochemical reactions atomic level. The technique has yielded exciting molecular insights into various biological processes including light sensing and energy conversion. However, achieve sufficient levels of activation within an optically dense crystal, high laser power densities are often used, which led ongoing debate extent photodamage may compromise interpretation...
Channelrhodopsins, light-gated cation channels, enable precise control of neural cell depolarization or hyperpolarization with light in the field optogenetics. This study integrates time-resolved serial crystallography and atomistic molecular dynamics (MD) simulations to resolve structural changes during C1C2 channelrhodopsin activation. Our observations reveal that within crystal environment, predominantly remains a light-activated state characteristics M
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 Conformational dynamics are essential for proteins to function. Here we describe how adapted time-resolved serial crystallography developed at X-ray lasers visualize protein motions using synchrotrons. We recorded the structural changes upon proton pumping in bacteriorhodopsin over 200 ms time. The snapshot from first 5 after photoactivation shows associated with release comparable quality previous laser experiments. From 10-15 onwards observe large additional rearrangements up 9 Å...
Abstract Die 8‐Demethyl‐8‐aminoriboflavin‐5′‐phosphat(AFP)‐Synthase RosB katalysiert die Schlüsselreaktion der Roseoflavinbiosynthese, Bildung von AFP aus Riboflavin‐5′‐phosphat (RP) und Glutamat. Zwischenprodukte dieser enzymatischen Reaktion sind 8‐Demethyl‐8‐formylriboflavin‐5′‐phosphat (OHC‐RP) 8‐Demethyl‐8‐carboxylriboflavin‐5′‐phosphat (HO 2 C‐RP). Um den schwierigen Austausch einer Methylgruppe am Aromaten gegen eine Aminogruppe zu verstehen, haben wir Struktur im Komplex mit OHC‐RP...