A5013529282- Photoreceptor and optogenetics research
- Photosynthetic Processes and Mechanisms
- Mass Spectrometry Techniques and Applications
- Enzyme Structure and Function
- Hemoglobin structure and function
- Metal-Catalyzed Oxygenation Mechanisms
- Receptor Mechanisms and Signaling
- Photochromic and Fluorescence Chemistry
- Microbial metabolism and enzyme function
- Molecular Communication and Nanonetworks
- Retinal Development and Disorders
- Spectroscopy and Quantum Chemical Studies
- Microbial bioremediation and biosurfactants
- Crystallization and Solubility Studies
- Porphyrin and Phthalocyanine Chemistry
- Neural dynamics and brain function
- Crystallography and molecular interactions
- Iron oxide chemistry and applications
- Heme Oxygenase-1 and Carbon Monoxide
- Genomics and Phylogenetic Studies
- X-ray Diffraction in Crystallography
- Metalloenzymes and iron-sulfur proteins
- Subcritical and Supercritical Water Processes
- Lipid Membrane Structure and Behavior
- Advanced X-ray Imaging Techniques
Paul Scherrer Institute
2018-2024
University of Essex
2014-2020
Colchester Institute
2016-2017
University of the West of England
2012
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...
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...
Significance A substantial number of known genetic disorders have their origin in mutations that cause misfolding or dysfunction G protein-coupled receptors (GPCRs). Pharmacological chaperones can rescue such mutant from the endoplasmic reticulum by stabilizing protein conformations support trafficking into target membrane. Rhodopsin-mediated retinitis pigmentosa is a disease might be targeted PCs. Here we present structure-based drug design approach to identify nonretinal compounds bind and...
The four Rep proteins of adeno-associated virus (AAV) orchestrate all aspects its viral life cycle, including transcription regulation, DNA replication, assembly, and site-specific integration the genome into human chromosome 19. All share a central SF3 superfamily helicase domain. In other members this domain is sufficient to induce oligomerization. However, in AAV (i.e. Rep40/Rep52) as shown by monomeric characteristic, not able mediate stable This observation led us hypothesize existence...
Powerful synergies are available from the combination of multiple methods to study proteins in crystalline form. Spectroscopies which probe same region crystal X-ray structures determined can give insights into redox, ligand and spin states complement information gained electron-density maps. The correct assignment protein redox is essential avoid misinterpretation structural data. This a particular concern for haem proteins, occupy wide range exquisitely sensitive becoming reduced by...
High-resolution crystal structures of enzymes in relevant redox states have transformed our understanding enzyme catalysis. Recent developments demonstrated that X-rays can be used, via the generation solvated electrons, to drive reactions crystals at cryogenic temperatures (100 K) generate `structural movies' reactions. However, a serious limitation these is protein conformational motion significantly supressed. Here, recently developed MSOX (multiple serial from one crystal) approach has...
Serial femtosecond crystallography (SFX) at X-ray free-electron lasers (XFELs) enables essentially radiation-damage-free macromolecular structure determination using microcrystals that are too small for synchrotron studies. However, SFX experiments often require large amounts of sample in order to collect highly redundant data where some the many stochastic errors can be averaged out determine accurate structure-factor amplitudes. In this work, capability Swiss laser (SwissFEL) was used...
ABSTRACT The adeno-associated virus (AAV) encodes four regulatory proteins called Rep. large AAV Rep Rep68 and Rep78 are essential factors required in almost every step of the viral life cycle. Structurally, they share two domains: a modified version AAA + domain that characterizes SF3 family helicases an N-terminal binds DNA specifically. combination these domains imparts extraordinary multifunctionality to work as initiators replication regulators transcription, addition their role during...
Nitrite coordination to heme cofactors is a key step in the anaerobic production of signaling molecule nitric oxide (NO). An ambidentate ligand, nitrite has potential coordinate via N- (nitro) or O- (nitrito) atoms manner that can direct its reactivity. Distinguishing nitro vs nitrito coordination, along with influence surrounding protein, therefore particular interest. In this study, we probed Fe(III) heme-nitrite Alcaligenes xylosoxidans cytochrome c′ (AXCP), an NO carrier excludes anions...
It is crucial to assign the correct redox and ligand states crystal structures of proteins with an active centre gain valid functional information prevent misinterpretation structures. Single-crystal spectroscopies, particularly when applied in situ at macromolecular crystallography beamlines, allow spectroscopic investigations identification reaction intermediates protein crystals during collection structural data. resonance Raman spectroscopy was carried out combination on Swiss Light...
Distal <italic>versus</italic> proximal binding of nitric oxide to haem is controlled <italic>via</italic> a ‘balance affinities’ kinetic mechanism.
Abstract 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. To trigger such protein-ligand interaction dynamics within human adenosine A 2A receptor, we designed seven photochemical affinity switches derived from anti-Parkinson’s istradefylline. In a rational approach based on UV/Vis spectroscopy, time-resolved...
Microbial nitrite reductases are denitrifying enzymes that a major component of the global nitrogen cycle. Multiple structures measured from one crystal (MSOX data) copper reductase at 240 K, together with molecular-dynamics simulations, have revealed protein dynamics type 2 site significant for its catalytic properties and entry exit solvent or ligands to active site. Molecular-dynamics simulations were performed using different protonation states key residues (Asp CAT His ) involved in...
<title>Abstract</title> Molecular photoswitches are versatile natural or synthetic molecules that undergo reversible conformational changes in response to light. In chemistry azobenzenes act as ubiquitous photoswitches<sup>1</sup> with applications ranging from opto-electronics<sup>2</sup>, over molecular machines<sup>3</sup> photopharmacology<sup>4</sup>. Their isomerization mechanism defines their properties and yet is controversially debated, the underlying ultrafast photochemistry...
Hydroxylamine (NH2OH or HA) is a redox-active nitrogen oxide that occurs as toxic intermediate in the oxidation of ammonium by nitrifying and methanotrophic bacteria. Within containing environments, HA generated ammonia monooxygenase (nitrifiers) methane (methanotrophs). Subsequent catalyzed heme proteins, including cytochromes P460 multiheme hydroxylamine oxidoreductases, former contributing to emissions N2O, an ozone-depleting greenhouse gas. A heme–HA complex also proposed reduction...
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...
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