A5022389520- Receptor Mechanisms and Signaling
- Photoreceptor and optogenetics research
- Enzyme Structure and Function
- Monoclonal and Polyclonal Antibodies Research
- Lipid Membrane Structure and Behavior
- Mass Spectrometry Techniques and Applications
- Retinal Development and Disorders
- Protein Structure and Dynamics
- Advanced Fluorescence Microscopy Techniques
- Amino Acid Enzymes and Metabolism
- RNA and protein synthesis mechanisms
- Neuropeptides and Animal Physiology
- Advanced Electron Microscopy Techniques and Applications
- Chemokine receptors and signaling
- Photosynthetic Processes and Mechanisms
- Microbial Metabolic Engineering and Bioproduction
- Computational Drug Discovery Methods
- Neuroscience and Neuropharmacology Research
- Glycosylation and Glycoproteins Research
- Metabolomics and Mass Spectrometry Studies
- Ion channel regulation and function
- SARS-CoV-2 and COVID-19 Research
- Long-Term Effects of COVID-19
- Advanced X-ray Imaging Techniques
- Force Microscopy Techniques and Applications
Paul Scherrer Institute
2015-2024
China Medical University
2019
ETH Zurich
2015
Universität Hamburg
2015
Center for Free-Electron Laser Science
2015
University of Cambridge
2013
Max Planck Institute of Biophysics
2005-2011
Medical University of Vienna
2010
Max Planck Society
2005
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...
The structure of CCR5 in complex with [6P4]CCL5 and the G i protein reveals its activation mechanism.
X-ray diffraction patterns from two-dimensional (2-D) protein crystals obtained using femtosecond pulses an free-electron laser (XFEL) are presented. To date, it has not been possible to acquire transmission individual 2-D due radiation damage. However, the intense and ultrafast generated by XFEL permit a new method of collecting data before sample is destroyed. Utilizing diffract-before-destroy approach at Linac Coherent Light Source, Bragg was acquired better than 8.5 Å resolution for two...
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...
One of the largest membrane protein families in eukaryotes are G protein-coupled receptors (GPCRs). GPCRs modulate cell physiology by activating diverse intracellular transducers, prominently heterotrimeric proteins. The recent surge structural data has expanded our understanding GPCR-mediated signal transduction. However, many aspects, including existence transient interactions, remain elusive. We present cryo-EM structure light-sensitive GPCR rhodopsin complex with Gi. Our density map...
Highlights•The head contains nicastrin ectodomain and overhangs a solute-accessible cavity in base•The base has central channel lateral cleft (putative substrate docking site)•Inhibitors close the rotate head, blocking accessSummaryPresenilin-mediated endoproteolysis of transmembrane proteins plays key role physiological signaling pathogenesis Alzheimer disease some cancers. Numerous inhibitors have been found via library screens, but their structural mechanisms remain unknown. We used...
Activation and inactivation of voltage-gated sodium channels (Navs) are well studied, yet the molecular mechanisms governing channel gating in membrane remain unknown. We present two conformations a Nav from Caldalkalibacillus thermarum reconstituted into lipid bilayers one crystal at 9 Å resolution based on electron crystallography. Despite voltage sensor arrangement identical with that activated form, we observed distinct pore domain structures: prominent form relatively open inner gate...
Abstract Effective public-health measures and vaccination campaigns against SARS-CoV-2 require granular knowledge of population-level immune responses. We developed a Tripartite Automated Blood Immunoassay (TRABI) to assess the IgG response ectodomain receptor-binding domain spike protein as well nucleocapsid SARS-CoV-2. used TRABI for continuous seromonitoring hospital patients healthy blood donors (n=72’222) in canton Zurich from December 2019 2020 (pre-vaccine period). Seroprevalence...
The binding of imidazolium salts to cucurbit[8]uril, CB[8], triggers a stepwise self-assembly process with semiflexible polymer chains and crystalline nanostructures as early- late-stage species, respectively. In such process, which involves the crystallization host–guest complexes, guest plays critical role in directing toward desirable morphologies. These include platelet-like aggregates two-dimensional (2D) fibers, which, moreover, exhibit viscoelastic lyotropic properties. Our...
Effective public health measures against SARS-CoV-2 require granular knowledge of population-level immune responses. We developed a Tripartite Automated Blood Immunoassay (TRABI) to assess the IgG response three proteins. used TRABI for continuous seromonitoring hospital patients and blood donors (n = 72′250) in canton Zurich from December 2019 2020 (pre-vaccine period). found that antibodies waned with half-life 75 days, whereas cumulative incidence rose 2.3% June 12.2% mid-December 2020. A...
Abstract In biological and biomedical research the focus progressively moves towards difficult human proteins, which often can only be expressed in higher eukaryotic cells. Nuclear magnetic resonance (NMR) could contribute significantly to understanding of important proteins as it is one most information-rich methods: allows studying structure, function dynamics biomolecules and, importantly, their interactions with natural ligands or drugs. However, exploit full potential NMR, must isotope...
X-ray free-electron lasers (XFELs) offer a new avenue to the structural probing of complex materials, including biomolecules. Delivery precious sample XFEL beam is key consideration, as interest must be serially replaced after each destructive pulse. The fixed-target approach delivery involves depositing samples on thin-film support and subsequent serial introduction via translating stage. Some classes biological two-dimensional protein crystals, introduced supports, they require flat...
Membrane proteins arranged as two-dimensional crystals in the lipid environment provide close-to-physiological structural information, which is essential for understanding molecular mechanisms of protein function. Previously, X-ray diffraction from individual did not represent a suitable investigational tool because radiation damage. The recent availability ultrashort pulses free-electron lasers (XFELs) has now provided means to outrun Here, we report on measurements performed at Linac...
Structural information of the different conformational states two prototypical light-sensitive membrane proteins, bacteriorhodopsin and rhodopsin, has been obtained in past by X-ray cryo-crystallography cryo-electron microscopy. However, these methods do not allow for structure determination most intermediate conformations. Recently, potential X-Ray Free Electron Lasers (X-FELs) tracking dynamics light-triggered processes pump-probe serial femtosecond crystallography demonstrated using...
Resistance to the current therapies is main clinical challenge in treatment of lethal metastatic prostate cancer (mPCa). Developing novel therapeutic approaches with effective regimes and minimal side effects for this fatal disease remain a priority study. In present study, we demonstrated that traditional Chinese medicine, quality-assured Ganoderma tsugae ethanol extract (GTEE), significantly suppressed cell growth capability caused cycle arrest through decreasing expression cyclins mPCa...
In this work, we examine methyl nuclear magnetic resonance (NMR) spectra of the methionine ε-[13CH3] labelled thermostabilized β1 adrenergic receptor from turkey in association with a variety different effectors, including mini-Gs and nanobody 60 (Nb60), which have not been previously studied complex (β1AR) by NMR. Complexes pindolol Nb60 induce highly similar inactive states receptor, closely resembling resting state conformational ensemble. We show that, upon binding or 80 (Nb80), large...
Significance G protein–coupled receptors (GPCRs) are important pharmaceutical targets for the treatment of a broad spectrum diseases. Upon ligand binding, GPCRs initiate intracellular signaling pathways by interacting with partner proteins. Assays that quantify interplay between binding and initiation downstream cascades critical in early stages drug development. We have developed high-throughput mass spectrometry method to unravel GPCR–protein complex demonstrated its use three provide...
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...
Previous proof-of-concept measurements on single-layer two-dimensional membrane-protein crystals performed at X-ray free-electron lasers (FELs) have demonstrated that the collection of meaningful diffraction patterns, which is not possible synchrotrons because radiation-damage issues, feasible. Here, results obtained from analysis a thousand single-shot, room-temperature FEL images bacteriorhodopsin mutant are reported in detail. The high redundancy boosts intensity signal-to-noise ratio, so...
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....