A5074330837- Receptor Mechanisms and Signaling
- Photoreceptor and optogenetics research
- Crystallization and Solubility Studies
- X-ray Diffraction in Crystallography
- Crystallography and molecular interactions
- Mass Spectrometry Techniques and Applications
- Retinal Development and Disorders
- Protein Kinase Regulation and GTPase Signaling
- Hereditary Neurological Disorders
- Neuroscience and Neuropharmacology Research
- Enzyme Structure and Function
- Prion Diseases and Protein Misfolding
- Monoclonal and Polyclonal Antibodies Research
- Protein Structure and Dynamics
- Biochemical and Structural Characterization
- RNA Research and Splicing
- Mesoporous Materials and Catalysis
- Synthesis and properties of polymers
- Silicone and Siloxane Chemistry
- Neurobiology and Insect Physiology Research
- Advanced Fluorescence Microscopy Techniques
- Neurological diseases and metabolism
- Metabolism, Diabetes, and Cancer
- Retinopathy of Prematurity Studies
- Neuropeptides and Animal Physiology
Paul Scherrer Institute
2018-2024
Molecular Research Institute
2020
Material (Belgium)
1999
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...
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...
Light-sensitive G protein-coupled receptors (GPCRs)-rhodopsins-absorb photons to isomerize their covalently bound retinal, triggering conformational changes that result in downstream signaling cascades. Monostable rhodopsins release retinal upon isomerization as opposed the bistable "reisomerize" absorption of a second photon. Understanding mechanistic differences between these light-sensitive GPCRs has been hindered by scarcity recombinant models latter. Here, we reveal high-resolution...
Abstract Animal vision depends on opsins, a category of G protein-coupled receptor (GPCR) that achieves light sensitivity by covalent attachment to retinal. Typically binding as an inverse agonist in the 11-cis form, retinal photoisomerizes all-trans isomer and activates receptor, initiating downstream signaling cascades. Retinal bound bistable opsins isomerizes back state after absorption second photon, inactivating receptor. Bistable are essential for invertebrate non-visual perception...
Animal vision depends on opsins, a category of G protein-coupled receptor (GPCR) that achieves light sensitivity by covalent attachment to retinal. Typically binding as an inverse agonist, 11-cis retinal photoisomerizes the all-trans isomer and activates receptor, initiating downstream signaling cascades. Retinal bound bistable opsins isomerizes back state after absorption second photon, inactivating receptor. Bistable are essential for invertebrate nonvisual perception across animal...
ABSTRACT Heterotrimeric G proteins play a central role in cellular signaling, acting as switchable molecular regulators. Consequently, pharmacological agents to control protein activity are of utmost importance advance our understanding this signal transduction system. The natural depsipeptides FR900359 (FR) and YM-254890 (YM) two highly specific widely used inhibitors heterotrimeric Gq/11 proteins. These compounds have traditionally been understood inhibit GDP dissociation by preventing the...
The adhesion G-protein coupled receptor Adgrg6 (formerly Gpr126) is instrumental in the development, maintenance and repair of peripheral nervous system myelin. prion protein (PrP) a potent activator could be used as potential therapeutic agent treating demyelinating dysmyelinating diseases. We designed dimeric Fc-fusion comprising myelinotrophic domain PrP (FT 2 Fc), which activated vitro exhibited favorable pharmacokinetic properties for vivo treatment neuropathies. While chronic FT Fc...
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....
Label free, biosensor-based screening approaches can be easily adapted for testing interactions of arrestins with different GPCRs.
Abstract Arrestin-1 desensitizes the activated and phosphorylated photoreceptor rhodopsin by forming transient rhodopsin−arrestin-1 complexes that eventually decay to opsin, retinal arrestin-1. Via a multi-dimensional screening setup, we identified combined arrestin-1 mutants form lasting with light-activated in harsh conditions, such as high ionic salt concentration. Two quadruple mutants, D303A + T304A E341A F375A R171A share similar heterologous expression thermo-stability levels wild...
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...
The selective G q inhibitor FR900359 was probed by NMR spectroscopy bound to the membrane-anchored heterotrimer, revealing its mode of membrane interaction, protein binding and allosteric response.
Abstract The adhesion G-protein coupled receptor Adgrg6 (formerly Gpr126) is instrumental in the development, maintenance and repair of peripheral nervous system myelin. prion protein (PrP) a potent activator could be used as potential therapeutic agent treating demyelinating dysmyelinating diseases. We designed dimeric Fc-fusion comprising myelinotrophic domain PrP (FT 2 Fc), which activated vitro exhibited favorable pharmacokinetic properties for vivo treatment neuropathies. While chronic...
The key to determining crystal structures of membrane protein complexes is the quality sample prior crystallization. In particular, choice detergent critical, because it affects both stability and monodispersity complex. We recently determined structure an active state bovine rhodopsin coupled engineered G protein, mini-Go, at 3.1 Å resolution. Here, we detail procedure for optimizing preparation rhodopsin–mini-Go Dark-state was prepared in classical neopentyl glycol (NPG) detergents,...
Abstract G protein-coupled receptors (GPCRs) are the largest class of integral membrane proteins and represent key targets for pharmacological research. GPCRs modulate cell physiology by engaging activating a diversity intracellular transducers, prominently heterotrimeric proteins, but also protein-receptor kinases (GRKs) arrestins. The recent surge in number structures GPCR-G protein complexes has expanded our understanding recognition GPCR-mediated signal transduction. However, many...
The key to determining crystal structures of membrane protein complexes is the quality sample prior crystallization. In particular, choice detergent critical, because it affects both stability and monodispersity complex. We recently determined structure an active state bovine rhodopsin coupled engineered G protein, mini-Go, at 3.1 Å resolution. Here, we detail procedure for optimizing preparation rhodopsin–mini-Go Dark-state was prepared in classical neopentyl glycol (NPG) detergents,...