Refine
Has Fulltext
- yes (305)
Is part of the Bibliography
- yes (305)
Year of publication
Document Type
- Journal article (196)
- Doctoral Thesis (102)
- Preprint (5)
- Master Thesis (1)
- Report (1)
Keywords
- Thrombozyt (19)
- platelets (17)
- ischemic stroke (10)
- Maus (9)
- Thrombose (8)
- platelet (8)
- gephyrin (7)
- platelet activation (6)
- GPVI (5)
- Microscopy (5)
- Signaltransduktion (5)
- blood (5)
- glycoprotein VI (5)
- platelet aggregation (5)
- thrombo-inflammation (5)
- Biologie (4)
- DNS-Reparatur (4)
- Drosophila melanogaster (4)
- Fluoreszenzmikroskopie (4)
- Mice (4)
- Mikroskopie (4)
- Platelets (4)
- Rezeptor (4)
- Staphylococcus aureus (4)
- Taufliege (4)
- Thrombosis (4)
- X-ray crystallography (4)
- actin (4)
- expression (4)
- integrins (4)
- megakaryocytes (4)
- phosphorylation (4)
- thrombosis (4)
- Arteriosklerose (3)
- Arzneimitteldesign (3)
- Dendritische Zelle (3)
- Diabetes mellitus (3)
- Drosophila (3)
- Entzündung (3)
- Fettsäurestoffwechsel (3)
- GPCR (3)
- Genexpression (3)
- HUWE1 (3)
- Hämostase (3)
- Inflammation (3)
- Megakaryozyt (3)
- Metalloproteinasen (3)
- NRF2 (3)
- Platelet (3)
- Proteinkinase D (3)
- Rac1 (3)
- Transkriptionsfaktor (3)
- Typ 1 (3)
- Ubiquitin (3)
- cancer (3)
- cell wall (3)
- crystal structure (3)
- differentiation (3)
- escherichia coli (3)
- flow cytometry (3)
- fluorescence imaging (3)
- fluorescence microscopy (3)
- inflammation (3)
- lipid bilayer (3)
- macrophages (3)
- mechanisms (3)
- protein (3)
- thrombin (3)
- tumorigenesis (3)
- AFM (2)
- ATF4 (2)
- Agent (2)
- Aorta (2)
- Apoptosis (2)
- Blut-Hirn-Schranke (2)
- Breast-tumors (2)
- C. elegans (2)
- C/EBP (2)
- CML (2)
- CXCR4 (2)
- Caenorhabditis elegans (2)
- Calcium (2)
- Cancer (2)
- Cryo-EM (2)
- Cytomegalie-Virus (2)
- Cytoskeleton (2)
- DNA repair (2)
- Differenzierung (2)
- Einzelmolekülmikroskopie (2)
- Expansion Microscopy (2)
- FRET (2)
- Fettsäurebiosynthese (2)
- Fluorescence (2)
- Fluoreszenz (2)
- G protein-coupled receptors (2)
- GABA (2)
- GABAA receptors (2)
- Gephyrin (2)
- Helicasen (2)
- Herzhypertrophie (2)
- Hirnhautentzündung (2)
- Inhibition (2)
- Inhibitorische Synapse (2)
- Inhibitory synapse (2)
- Interleukin 6 (2)
- Invasion (2)
- Kollagen (2)
- Kristallographie (2)
- Kristallstruktur (2)
- LASP1 (2)
- Learning and memory (2)
- Lung cancer (2)
- MIZ1 (2)
- MYC (2)
- Megakaryocyte (2)
- Megakaryopoese (2)
- Meningitis (2)
- Metabolism (2)
- Molekulargenetik (2)
- Muskelzelle (2)
- Nude-mice (2)
- OSMR (2)
- Obesity (2)
- Oncostatin M (2)
- PIP2 (2)
- Peptidsynthese (2)
- Phagozytose (2)
- Phosphorylierung (2)
- Plasmamembran (2)
- Plättchen (2)
- Pneumolysin (2)
- Protein (2)
- Protein p53 (2)
- Protein p73 (2)
- Regulation (2)
- Rhabdomyosarkom (2)
- Rho GTPase (2)
- SOCE (2)
- Schlaganfall (2)
- Smad (2)
- T-Lymphozyt (2)
- T-cells (2)
- TFIIH (2)
- Therapy (2)
- Toxin (2)
- Type 1 Diabetes (2)
- Zelle (2)
- Zellmigration (2)
- Zellskelett (2)
- Zelltod (2)
- activation (2)
- artemisinin (2)
- atherosclerosis (2)
- atomic-force microscopy (2)
- binding (2)
- biosensors (2)
- blood coagulation (2)
- bone marrow (2)
- brain (2)
- breast-tumors (2)
- cell biology (2)
- cell signalling (2)
- chloroquine (2)
- coagulation (2)
- collagens (2)
- collybistin (2)
- crystallography (2)
- dSTORM (2)
- dendritic cells (2)
- diazepam (2)
- domain (2)
- drug discovery (2)
- enzymes (2)
- extracellular domain (2)
- factor XII (2)
- fluorescence resonance energy transfer (FRET) (2)
- gene expression (2)
- glycine receptor (2)
- gp130 (2)
- human (2)
- hydrodynamics (2)
- inhibitory neurotransmission (2)
- inhibitory postsynapse (2)
- interactome (2)
- ischemic penumbra (2)
- isothermal titration calorimetry (2)
- lipid bilayer membrane (2)
- matrix protein porin (2)
- melanoma (2)
- membrane (2)
- membrane potential (2)
- meningitis (2)
- metastasis (2)
- miRNS (2)
- mice (2)
- middle cerebral artery occlusion (2)
- molecular biology (2)
- mouse (2)
- mouse model (2)
- mouse models (2)
- murine (2)
- neuroinflammation (2)
- neuronal dendrites (2)
- neurons (2)
- nucleotide excision repair (2)
- nucleotide excision-repair (2)
- nude-mice (2)
- oncolytic viruses (2)
- p34 (2)
- p44 (2)
- p53 (2)
- p63 (2)
- p73 (2)
- pathway (2)
- phagocytosis (2)
- phagosome maturation (2)
- phospholipase D (2)
- platelet receptors (2)
- porin (2)
- recognition (2)
- rheumatoid arthritis (2)
- signaling (2)
- stroke (2)
- structural basis (2)
- structure based drug design (2)
- sulfur (2)
- synapses (2)
- synaptic plasticity (2)
- therapy (2)
- thermodynamics (2)
- thrombopoiesis (2)
- translocation (2)
- tumors (2)
- type VII secretion system (2)
- ubiquitin (2)
- vision (2)
- vitamin B6 (2)
- 1,25-dihydroxyvitamin D\(_{3}\) (1)
- 2-photon microscopy (1)
- 3D printing (1)
- 7,8-dihydroxyflavone (7,8-DHF) (1)
- 7-Dehydrocholesterol (1)
- 8-oxoguanine (1)
- AAA (1)
- AAA+ ATPase p97 (1)
- ADAM10 (1)
- ADP-ribosyltransferases (1)
- AGT (1)
- AKT1 (1)
- AMP-activated protein kinase (AMPK) (1)
- AMP‐activated protein kinase (1)
- ATGL (1)
- Abszission (1)
- Acetylation (1)
- Acetylierung (1)
- Ackerschmalwand (1)
- Actin (1)
- Actin binding proteins (1)
- Actin cytoskeleton-related protein (1)
- Actin-bindende Proteine (1)
- Activation (1)
- Active zone (1)
- Adapterprotein (1)
- Adipokine (1)
- Adipositas (1)
- Adrenalin (1)
- Affinity probe (1)
- Aktive Zone (1)
- Allgemeine Zelle (1)
- Anthrax Toxin (1)
- Anti-Gehirn Autoantikörper (1)
- Antikörper (1)
- Antithrombotics (1)
- Antithrombotika (1)
- Antithrombotikum (1)
- Archaea (1)
- Archaebakterien (1)
- Arp2/3 complex (1)
- Art (1)
- Arterial water (1)
- Arterielles Blut (1)
- Aspergillus fumigatus (1)
- Astrozyt (1)
- Atherosclerosis (1)
- Atriales natriuretisches Hormon (1)
- Autoantigen (1)
- Autoantikörper (1)
- Autoinhibition (1)
- Automatische Biegewinkelmessung (1)
- Autophagie (1)
- BCR‐ABL (1)
- BIN2 (1)
- BMP (1)
- BRAF (1)
- BTB domain (1)
- BV-2 (1)
- Bacillus anthracis (1)
- Bacillus subtilis (1)
- Bacterial Toxins (1)
- Bakteriengift (1)
- Basen-Exzisionsreparatur (1)
- Bcl-2 (1)
- Behavior (1)
- Beige adipocytes (1)
- Bericht (1)
- Biegewinkel (1)
- Bildbearbeitung (1)
- Bildgebung intakten Knochens (1)
- Bildverarbeitung (1)
- Bindungsprozess (1)
- Biochemie (1)
- Biochemistry (1)
- Biology (1)
- Biomarker (1)
- Black-lipid-bilayer (1)
- Blood-brain barrier (1)
- Blutgefäßsystem (1)
- Blutplättchen (1)
- Bone morphogenetic proteins (1)
- Bordetella pertussis (1)
- Brain (1)
- Burkholderia (1)
- Burkholderia pseudomallei (1)
- C2-toxin (1)
- CCR2 (1)
- CD coreceptors (1)
- CD11b+ myeloid cells (1)
- CD84 (1)
- CHAC1 (1)
- CIB1 (1)
- CLEC-2 (1)
- CLEC-2 ITAM (1)
- COVID-19 (1)
- COX-2 (1)
- COX2 expression (1)
- CVT (1)
- CXCL12-abundant reticular (CAR)-cells (1)
- CXCL4 (1)
- CXCL7 (1)
- Calcium signalling (1)
- Calcium-bindende Proteine (1)
- Carboxy-Terminus (1)
- Carcinogenese (1)
- Carcinoma (1)
- Caspr1 (1)
- Cation Homeostasis (1)
- Cell (1)
- Cell line (1)
- Cells (1)
- Central nervous system (1)
- Cerebral-ischemia (1)
- ChIP-sequencing (1)
- Chaetomium thermophilum (1)
- Chemerin (1)
- Chemerin processing (1)
- Chemotherapy (1)
- Chlamydia trachomatis (1)
- Cholesterin (1)
- Cholesterol (1)
- Chylomicron (1)
- Chylomicrons (1)
- Circadian rhythms (1)
- Clec16a (1)
- Clostridioides binary toxins (1)
- Clostridium-botulinum (1)
- Cluster (1)
- Coefficient (1)
- Collaborative Research Center (1)
- Collagen (1)
- Combination therapy (1)
- Component selectivity (1)
- Computerprogramm (1)
- Conditioning (1)
- Contactin 1 (1)
- Cortactin (1)
- Corynebacterium urealyticum (1)
- Cranial window (1)
- Crystal structure of MTR1 (1)
- Crystal-structure (1)
- Current noise-analysis (1)
- CyaA (1)
- Cyclophosphamide (1)
- Cys-loop receptor (1)
- Cytokine (1)
- DAMGO (1)
- DAPI staining (1)
- DHCR7 (1)
- DNA Repair (1)
- DNA damage (1)
- DNA metabolism (1)
- DNA origami (1)
- DNA structures (1)
- DNA-Reparatur (1)
- DNA-based nanostructures (1)
- DNA-processing enzymes (1)
- DNS-Bindung (1)
- Darm (1)
- Deletion (1)
- Dendra2 (1)
- Dendritic cells (1)
- Diabetes (1)
- Diabetes Typ 1 (1)
- Differentiation (1)
- Diphenylether (1)
- Disulfidbrücken (1)
- Diversifikation <Biologie> (1)
- Dopamine (1)
- Durchflusscytometrie (1)
- Dynamik (1)
- Dynamik von Membranrezeptoren (1)
- E1 (1)
- E2 (1)
- E3 enzyme (1)
- EHT1864 (1)
- ERK map kinease (1)
- ERK1/2 (1)
- ESAT‐6‐like secretion system (1)
- ESS (1)
- Edema factor (1)
- Einzelpartikelverfolgung (1)
- Enoyl-Reduktase (1)
- Enoyl-acyl-carrier-protein-Reductase (1)
- Entwicklung (1)
- Enzym (1)
- Enzyme Regulation (1)
- Epitop (1)
- Erlernte Hilflosigkeit (1)
- EsaA (1)
- Escherichia coli (1)
- Escherichia coli AlkA (1)
- Escherichia-coli (1)
- Experimental Biomedicine (1)
- Experimental stroke (1)
- Extracellular Vesicles (1)
- FANCM (1)
- FAS-II (1)
- FAT10ylation (1)
- FIONA (1)
- FMMs (1)
- FRAP (1)
- FRET sensors (1)
- FabI (1)
- FabV (1)
- FeS cluster (1)
- Ferroptose (1)
- Ferroptosis (1)
- Fettsäure-Synthase (1)
- Fgf-signalling (1)
- Fibroblast (1)
- Flippase (1)
- Flotillin (1)
- Fluorescence Microscopy (1)
- Fluorescence Resonance Energy Transfer (1)
- Fluorescence correlation spectroscopy (1)
- Fluorescence imaging with one nanometer accuracy (1)
- Fluorescence microscopy (1)
- Fluorescence resonance energy transfer (1)
- Fluorescent probes (1)
- Fluoreszenzsonde (1)
- Fluoreszmikroskopie (1)
- FoxO3 (1)
- Foxo1 (1)
- Fragiles X Syndrom (1)
- Frühdiagnostik (1)
- Förster Resonance Energy Transfer (1)
- Förster resonance energy transfer (1)
- G Protein-Coupled Receptors (1)
- G protein-coupled receptor (1)
- G-Protein gekoppelte Rezeptoren (1)
- G-protein coupled receptors (1)
- G-protein-coupled receptors (1)
- GABA(A) receptors (1)
- GABA-A receptor (1)
- GABAA (1)
- GBM (1)
- GFAP (1)
- GI-101A tumor xenografts (1)
- GIST (1)
- GLV-1 h153 (1)
- GLV-1H68 (1)
- GPCRs (1)
- GPVI shedding (1)
- GPX4 (1)
- GSH (1)
- GapN (1)
- Gelernte Hilflosigkeit (1)
- Genanalyse (1)
- Gene-expression (1)
- General Transcription Factor II H (1)
- Gerinnungsfaktor (1)
- Geruchswahrnehmung (1)
- Gleiten (1)
- Gliafaserprotein (1)
- Glycoprotein GPV (1)
- Glycoprotein VI (1)
- Glycoprotein hormone (1)
- Glykoprotein VI (1)
- Glykoprotein-Shedding (1)
- Glykoproteine (1)
- Glykosylasen (1)
- Gram-positive bacteria (1)
- Guanine nucleotide exchange factor (GEF) (1)
- Guanylatcyclase (1)
- Guanylyl cyclase A (1)
- Gα\(_{i1}\), Gα\(_{i2}\) and Gα\(_{i3}\) activation (1)
- H1 helix (1)
- H1-Helix (1)
- H2A histone family member X (H2AX) (1)
- HECT Ligase (1)
- HHV-6A (1)
- HIF1alpha (1)
- Head-injury (1)
- Heart (1)
- Helicase (1)
- Hemodynamic depression (1)
- Hemostasis (1)
- Hepatitis B Virus (1)
- Herpesvirus (1)
- Heubacillus (1)
- Hic-5 (1)
- High-throughput screening (1)
- Hirschsprung disease (1)
- Homöostase (1)
- Hypothalamus (1)
- Hypoxia (1)
- Hypoxie (1)
- IMA2.1 (1)
- ITAM (1)
- Identification (1)
- Immune Thrombocytopenia (1)
- Immunologische Synapse (1)
- Immunthrombozytopenie (1)
- In vivo (1)
- In vivo imaging (1)
- In-vivo (1)
- Inhibitor (1)
- Inhibitory-postsynapse (1)
- Insulin (1)
- Insulinsekretion (1)
- Integrine (1)
- Interleukin 17 (1)
- Intravascular coagulation (1)
- Intrazellulärraum (1)
- Invertebrate vision (1)
- Ion-channel (1)
- Iron-uptake (1)
- Ischemic stroke (1)
- JAQ1 (1)
- Jolly bodies (1)
- KEA (1)
- KEAP1 (1)
- KIT (1)
- Kardial Hypertrophy (1)
- Kardiovaskulär (1)
- Karzinomzellen (1)
- Kationen-Homöostase (1)
- Knochen-Morphogenese-Proteine (1)
- Knockout <Molekulargenetik> (1)
- Koagulation (1)
- Kollektive Invasion (1)
- Konditionierung (1)
- Kraftmikroskopie (1)
- Krebsforschung (1)
- Krebszelle (1)
- L-3,4-Dihydroxyphenylalanine-induced dyskinesia (1)
- LC-MS (1)
- LC3-associated phagocytosis (1)
- LC3-assoziierte Phagozytose (1)
- LIFR (1)
- Latrophilin (1)
- Learned Helplessness (1)
- Lebenszyklus (1)
- Leber-Metabolismus (1)
- Lentiviral transgenesis (1)
- Lernen und Gedächtnis (1)
- Lethal factor (1)
- Leukaemia-inhibitory factor (1)
- Lichtblattmikroskopie (1)
- Lichtscheibenmikroskopie (1)
- Lipid Rafts (1)
- Lipid bilayer-membranes (1)
- Lipide (1)
- Lipolysis (1)
- LpxC inhibitors (1)
- Lungenkrebs (1)
- Lymphom (1)
- M14 carboxypeptidasses (1)
- MAPK signaling (1)
- MAX (1)
- MGL (1)
- MRI reporter (1)
- MYCN (1)
- MYCNv (1)
- Magnetic-resonance (1)
- Malignant effusion (1)
- Maligne Transformation (1)
- Mass Spectrometry (1)
- Massenspektrometrie (1)
- Mast cells (1)
- Matrix protein porin (1)
- Mc4r (1)
- Mcl-1 (1)
- Mechanismus (1)
- Medicine (1)
- Medizin (1)
- Megakaryozytopoese (1)
- Membrane Receptor Dynamics (1)
- Membranglykoproteine (1)
- Membranrezeptor (1)
- Metabolismus (1)
- Metabotropic glutamate receptor (1)
- Metalloproteases (1)
- Metalloproteinase (1)
- Methyltransferase Ribozyme MTR1 (1)
- Microarray (1)
- Microvesicle (1)
- Mitose (1)
- Mittelkörper (1)
- Model (1)
- Molecular Biophysics (1)
- Molecular-weight heparin (1)
- Molekularbiologie (1)
- Molekulare Marker (1)
- Mouse (1)
- Mouse model (1)
- Multidrug-Resistenz (1)
- Multiphotonenmikroskopie (1)
- Multiproteinkomplex (1)
- Mushroom body (1)
- Muskelkontraktion (1)
- Myomesin (1)
- NADPH (1)
- NAP-2 (1)
- NF-KAPPA-B (1)
- NFκB-activation (1)
- NOD (1)
- Nanofabrication (1)
- Nanofabrikation (1)
- Ndrg1 (1)
- Neurobiologie (1)
- Neurofascin (1)
- Neurogenetik (1)
- Neurons (1)
- Neuroscience (1)
- Neurowissenschaften (1)
- Neutralisation <Chemie> (1)
- Nrf2 (1)
- Nuclease (1)
- Nucleasen (1)
- Nucleotide-Excision-Repair (1)
- Nukleotid-Exzisions-Reparatur (1)
- O6-alkylguanine-DNA alkyltransferase (1)
- OSC (1)
- Obscurin (1)
- Odor-feeding-time memory (1)
- Olig2 (1)
- Oncolytic vaccinia virus (1)
- Oncolytic virotherapy (1)
- OprO (1)
- OprP (1)
- Orai2 (1)
- P14ARF (1)
- P4-ATPase (1)
- PDE (1)
- PDI (1)
- PDXP inhibitors (1)
- PF4 (1)
- PKA signaling (1)
- PKD1 (1)
- PKR (1)
- PNA (peptide nucleic acid) (1)
- PTMs (1)
- PTPN22 (1)
- Parkinson Disease (1)
- Parkinson-Krankheit (1)
- Parkinsons disease (1)
- Peptidase inhibitor 16 (PI16) (1)
- Peptide (1)
- Perforine (1)
- Perfusion (1)
- Peyer's patch (1)
- Phenylalanine clamp (1)
- Phospholipase D (1)
- Phosphorylation (1)
- Pilzkörper (1)
- Plasmonic (1)
- Platelet activating Factor (1)
- Platelet activation (1)
- Platelet-Membranglykoprotein p62 (1)
- Plätchen aktivierung (1)
- Polkörper (1)
- Pore (1)
- Pore formation (1)
- Pore-formation (1)
- Porenbildung (1)
- Posttranslationale Änderung (1)
- Primärtumor (1)
- Profilierung (1)
- Protease inhibition (1)
- Protective antigen (1)
- Protein Disulfid Isomerase (1)
- Protein Kinase C (1)
- Protein Kinase D (1)
- Protein Kinase D 1 (1)
- Protein Kinase D2 (1)
- Protein kinase D1 (PKD1) (1)
- Protein kinase D3 (PKD3) (1)
- Proteinfaltung (1)
- Proteinkinase C (1)
- Proteins (1)
- Protoplasten (1)
- Ptpn22 (1)
- Purification (1)
- Quantifizierung (1)
- Quantitation (1)
- Quergestreifte Muskulatur (1)
- RARRES2 (1)
- RCK domain (1)
- RHO (1)
- RHO-associated kinease (1)
- RKIP (1)
- RNA (1)
- RNA interference (1)
- RNA polymerase II (1)
- RNA splicing (1)
- RNA structures (1)
- RNA-catalyzed RNA methylation (1)
- RNAi (1)
- RNS-Interferenz (1)
- ROCK (1)
- ROK-alpha (1)
- RSK (1)
- RTX-Toxins (1)
- Rac (1)
- Ranvier-Schnürring (1)
- Rasterkraftmikroskop (1)
- Rats (1)
- Ratte (1)
- RecQ4 (1)
- Receptor Preference (1)
- Receptor internalization (1)
- Receptor signaling (1)
- Recombinant vaccinia (1)
- Registrierung <Bildverarbeitung> (1)
- Regression (1)
- Remorin (1)
- Rheumatoide Arthritis (1)
- Rho GTPasen (1)
- Rho GTPasw (1)
- RhoGTPase (1)
- Rhodesain (1)
- Ribosom (1)
- Röntgenkristallographie (1)
- S-layer protein (1)
- S6KII RSK (1)
- SARS-CoV-2 (1)
- SDS polyacrylamide gel electrophoresis (1)
- SLC7A11 (1)
- SOC (1)
- SPT5 (1)
- SPT6 (1)
- STIM2 (1)
- SUPT5H (1)
- Salmonella-typhimurium (1)
- Schlafkrankheit (1)
- Secretion (1)
- Sepsis (1)
- Serotonin (1)
- Sezernierte Faktoren (1)
- Signaling (1)
- Single Particle Tracking (1)
- Sinusthrombose (1)
- Solid-phase peptide synthesis (1)
- Sonderforschungsbereich Transregio 240 (1)
- Spezifität (1)
- Spleen tyrosine kinase (1)
- Src family (1)
- Ssl1 (1)
- Staphylococcus aureus USA300 (1)
- Store-Operated (1)
- Streptococcus pneumoniae (1)
- Stress (1)
- Structural Biology (1)
- Struktur-basiertes Wirkstoff Design (1)
- Sugar-transport (1)
- Super-Resolution Microscopy (1)
- Super-resolution microsopy (1)
- Superhochauflösende Mikroskopie (1)
- Synapse (1)
- T cell (1)
- T cell development (1)
- T cell migration (1)
- T cell rezeptor transfer (1)
- T lymphocyte (1)
- T-Lymphozyten-Rezeptor (1)
- T-Zell-Rezeptor Transfer (1)
- T-Zellaktivierung (1)
- T-Zellhomöostase (1)
- T-Zellrezeptor (1)
- TGN1412 (1)
- TIG2 (1)
- TPp73 (1)
- TRRAP (1)
- Tagesrhythmus (1)
- Telomerase (1)
- Tfb4 (1)
- Thrombopoese (1)
- Thrombopoiesis (1)
- Thrombozytenaggregation (1)
- Thrombozytenfunktionsanalyse (1)
- Thrombozytopathie (1)
- Thrombozytopoese (1)
- Thrombus (1)
- Thrombus formation (1)
- Tissue (1)
- Tissue staining (1)
- Titin (1)
- Transcription (1)
- Transkription (1)
- Translokation (1)
- Transparent motion (1)
- Triclosan (1)
- Triton X 100 (1)
- Trypanosoma brucei (1)
- Trypanosomiasis (1)
- TspanC8 (1)
- Tuberkelbakterium (1)
- Tumorigenese (1)
- Tumorzelle (1)
- Type 1 diabetes (1)
- Type VIIb secretion system (1)
- UBA6 (1)
- UBE2S (1)
- UBE2Z (1)
- UBE3A (1)
- US22 gene family (1)
- USP (1)
- Ube2S (1)
- Ubiquitin-PA (1)
- Ubiquitin-Protein-Ligase (1)
- Ubiquitin-conjugating (E2) enzymes (1)
- Ubiquitin-conjugating enzyme (1)
- Ubiquitinierung (1)
- Ubiquitylation (1)
- Unique Selling Proposition (1)
- VACV (1)
- VEGF (1)
- VSMC (1)
- Vaccinia virus (1)
- Vascular endothelial Growth Factor (1)
- Vaskuläre Integrität (1)
- Verhalten (1)
- Verweildauer (1)
- Vesikelbildung (1)
- Visual attention (1)
- Visuelle Aufmerksamkeit (1)
- Von-Willebrand-factor (1)
- WDR5 (1)
- Warsaw breakage syndrome (1)
- Wilms tumor (1)
- Wirkstoff (1)
- X-Ray Chrystallography (1)
- X-ray Crystallography (1)
- XPD (1)
- XPD helicase (1)
- Xeroderma pigmentosum (1)
- Xiphophorus (1)
- X‐ray crystallography (1)
- Yersinia (1)
- Yersinia pestis (1)
- Zebrabärbling (1)
- Zebrafisch (1)
- Zebrafish (1)
- Zelldifferenzierung (1)
- Zellkern (1)
- Zelllinie (1)
- Zelltransport (1)
- Zellwand (1)
- Zentralnervensystem (1)
- Zielstruktur (1)
- Zinc (1)
- Zink-Finger-Proteine (1)
- Zytoskelett (1)
- abscission (1)
- accelerated atherosclerosis (1)
- accumulation (1)
- acetylsalicylic acid (1)
- actins (1)
- activating transcription factor 4 (ATF4) (1)
- active zone (1)
- acute graft-versus host disease (1)
- acute graft-versus-host disease (1)
- acute myeloid leukemia (1)
- acute slices (1)
- adaptor protein Swiprosin-1/EFhd2 (1)
- additive manufacturing (1)
- adenylate cyclase toxin (1)
- adenylyl cyclase (1)
- adhesion GPCR (1)
- adipocyte (1)
- adipocytes (1)
- adipose (1)
- adrenergic receptor (1)
- adsorption (1)
- aggregation (1)
- alkene-alkyne [2+2] photocycloaddition (1)
- alkylation damage (1)
- alloreactive T cells (1)
- allosterism (1)
- amino acid sequence (1)
- aminoquinolinium salts (1)
- ampa receptors (1)
- ancistrocladinium A (1)
- angiogenesis (1)
- animal behavior (1)
- animal models (1)
- animal-model (1)
- animals (1)
- anions (1)
- antagonists (1)
- anthrax (1)
- anti-brain autoantibodies (1)
- anti-inflammatory (1)
- antibacterial activity (1)
- antibiotics (1)
- antibody (1)
- antibody/autoantibody (1)
- antigen processing and recognition (1)
- antioxidant function (1)
- aorta (1)
- apolipoprotein E (1)
- apoptose (1)
- apoptosis (1)
- arachidonic acid metabolic network (1)
- aspergillus fumigatus (1)
- astrocytes (1)
- ataxia teleagiectasia mutated (ATM) (1)
- atheriosclerotic lesions (1)
- atomic force microscopy (1)
- atomic force microscopy (AFM) (1)
- atopic dermatitis (1)
- atrial natriuretic peptide (1)
- attention (1)
- autoantibodies (1)
- autoantibody (aAb) (1)
- autoantigen (1)
- autoimmunity (1)
- autoinhibition (1)
- axonal transport (1)
- bacteria (1)
- bacterial fatty-acid biosynthesis (1)
- bacterial meningitis (1)
- base excision repair (1)
- base pairs (1)
- bead models (1)
- behavioral conditioning (1)
- beige adipocytes (1)
- beta (1)
- beta cell function (1)
- biased signaling (1)
- binding components (1)
- binding mode (1)
- biochemical characterization (1)
- bioconjugation (1)
- biological techniques (1)
- biomarkers (1)
- biomedicine (1)
- biopharmaceuticals (1)
- biosensor (1)
- black lipid bilayer (1)
- blood brain barrier (1)
- blood flow (1)
- blood platelets (1)
- blue native polyacrylamide gel electrophoresis (1)
- bone morphogenetic protein (1)
- bone-marrow (1)
- borrelia burgdorferi (1)
- boundary cap (1)
- brain damage (1)
- breast cancer (1)
- bulky damages recognition (1)
- cAMP (1)
- cAMP signaling (1)
- calcitonin gene-related peptide (1)
- calcium (1)
- calmodulin (1)
- cancer biology (1)
- cancer metabolism (1)
- cancer therapy (1)
- cancer treatment (1)
- carboxy terminus (1)
- carcinoma (1)
- carcinoma cells (1)
- cardiac hypertrophy (1)
- cardiovascular (1)
- cardiovascular disease (1)
- cascade (1)
- cell (1)
- cell corpse clearance (1)
- cell membranes (1)
- cell migration (1)
- cell proliferation (1)
- cell staining (1)
- cell wall channel (1)
- cell walls (1)
- cell-autonomous defense (1)
- cell-cycle arrest (1)
- cell-line (1)
- cells (1)
- cellular neuroscience (1)
- cellular stress response (1)
- cellular-trafficking (1)
- cerebellum (1)
- cerebrovascular diseases (1)
- cerebrovascular disorders (1)
- channel (1)
- chemical libraries (1)
- chemokine (1)
- chemokines (1)
- chemotherapy (1)
- cholesterol (1)
- cholesterol-dependent cytolysin (1)
- cholinergic interneurons (1)
- chronic heart failure (1)
- chronic kidney disease (1)
- chronophin (1)
- chylomicron (1)
- ciliary neurotrophic factor (1)
- click-chemistry (1)
- clinical neurology (1)
- coagulation system (1)
- coffin-lowry-syndrome (1)
- collective invasion (1)
- combinatorial libraries (1)
- complex (1)
- complex stability (1)
- computational docking (1)
- conditioned response (1)
- conformational activation (1)
- conformational auto-epitope (1)
- conjugation (1)
- conservation (1)
- conserving surgery (1)
- containing GABA(A) receptors (1)
- controllability (1)
- convergent extension movements (1)
- corticosteroids and cyclophosphamide (1)
- corynebacteria (1)
- corynebacterium diphtheriae (1)
- cultured hippocampal-neurons (1)
- cyclic AMP (1)
- cyclic peptides/cyclopeptides (1)
- cyclopeptide therapy (1)
- cysteine (1)
- cytokine release (1)
- cytomegaloviren (1)
- cytomegalovirus (1)
- cytoskeleton (1)
- cytotoxic T cells (1)
- dCIRL (1)
- damage responses (1)
- damaged DNA (1)
- deficient mice (1)
- demography (1)
- dendric cells (1)
- dendritic cell (1)
- dendritic growth (1)
- dendritic spines (1)
- dentate gyrus (1)
- detergents (1)
- deubiquitinase (1)
- developmental cycle (1)
- diabetes (1)
- diacylglycerol (DAG) (1)
- diet (1)
- dimeric peptide (1)
- dimerization (1)
- direct muss spectrometric profiling (1)
- directionality (1)
- disulfide bonds (1)
- dogs (1)
- domain swapping (1)
- dominant (1)
- dopa-induced dyskinesia (1)
- dopamine (1)
- dopamine transporters (1)
- dopaminergics (1)
- double knockout mice (1)
- drosophila melanogaster (1)
- drug development (1)
- drug repurposing (1)
- drug target (1)
- dunce (1)
- ectodomain cleavage (1)
- electrohydrodynamics (1)
- electron cryo microscopy (1)
- electron cryo-microscopy (1)
- electron microscopy (1)
- elongation rate (1)
- embryos (1)
- end-stage renal failure (1)
- endocannabinoid release (1)
- endothelial cell (1)
- endothelial cell interactions (1)
- endothelial growth-factor (1)
- enoyl reductase (1)
- enoyl-ACP reductase (1)
- enoyl-ACP reductase inhibitors (1)
- envelopment (1)
- enzyme (1)
- enzyme activator (1)
- enzyme mechanism (1)
- enzyme mechanisms (1)
- epithelial-mesenchymal transition (1)
- epitope (1)
- essential genes (1)
- exchange factor collybistin (1)
- experimental autoimmune encephalomyelitis (1)
- extracellular vesicle (1)
- eye movements (1)
- eyes (1)
- factor Sox10 (1)
- factor-I (1)
- fat absorption (1)
- fatty acid biosynthesis (1)
- fatty acid synthesis (1)
- fentanyl (1)
- ferroptosis (1)
- fetal liver (1)
- fibrin (1)
- fibrinogen (1)
- field flow fractionation (1)
- fine-mapping (1)
- flippase (1)
- flotillin (1)
- fluorescence (1)
- fluorescence lifetime imaging microscopy (1)
- fluorescence resonance energy transfer (1)
- fluorescence-detected sedimentation (1)
- fluorescent probes (1)
- fluorescent protein (1)
- fosmidomycin (1)
- fostamatinib (1)
- fractalkine (1)
- fragment screening (1)
- friut fly behaviour (1)
- fusion and fission (1)
- gamma-aminobutyric-acid (1)
- gastric cancer (1)
- gel electrophoresis (1)
- gene (1)
- gene therapy (1)
- general transcription factor IIH (TFIIH) (1)
- genetic code expansion (1)
- genome integrity (1)
- genomic stability (1)
- germline mutation (1)
- glutamate (1)
- glutamate receptor (1)
- glv-1h68 (1)
- glycine (1)
- glycine receptor (GlyR) (1)
- glycine transporter 2 (1)
- glycolytic flux control (1)
- glycoprotein GPV (1)
- glycoprotein Ib (1)
- glycoprotein Ibα (1)
- glycoprotein receptor Ib (1)
- glycoprotein receptor Ibα (1)
- glycoprotein-shedding (1)
- glycosylation (1)
- glyvine uptake (1)
- gram negative bacteria (1)
- gram-negative bacteria (1)
- granule cells (1)
- growth regulator (1)
- guanine nucleotide exchange factor (GEF) (1)
- gut microbiome (1)
- hTERT (1)
- haemostasis (1)
- haloacid dehalogenase (1)
- helicase (1)
- hematopoiesis (1)
- hemodialysis (1)
- hemostasis (1)
- hemostasis, (1)
- hepatitis B core protein (1)
- hepatitis B virus (1)
- hippocampus (1)
- histologic diversity (1)
- histology (1)
- homeostasisIon channels (1)
- homoFRET (1)
- homology modeling (1)
- hormone receptors (1)
- human cells (1)
- human interleukin-4 (1)
- human sodium iodide symporter (hNIS) (1)
- humanized tumor (1)
- humans (1)
- hybrid imaging (1)
- hybridomas (1)
- hypercholeterolemia (1)
- hyperekplexia (1)
- hyperlipedemic mice (1)
- hypertonic solution (1)
- identification (1)
- image analysis (1)
- imaging and sensing (1)
- imiquimod (1)
- immune cell function (1)
- immune cell recruitment (1)
- immune evasion (1)
- immunohistochemistry techniques (1)
- immunological synapse (1)
- immunoprecipitation (1)
- in vitro (1)
- in vitro and in vivo thrombus formation (1)
- in vivo (1)
- in vivo Modelle (1)
- in vivo cell tracking (1)
- in vivo imaging (1)
- in vivo models (1)
- in-vitro (1)
- in-vitro propagation (1)
- in-vivo (1)
- inactivation (1)
- increases atherosclersosis (1)
- indoleamine 2,3-dioxygenase (1)
- indoxyl sulfate (1)
- inflammatory sites (1)
- inhibition (1)
- inhibitor residence time (1)
- inhibitors (1)
- inhibitory synapse (1)
- innate immune response (1)
- insect flight (1)
- insulin (1)
- intact bone imaging (1)
- integrated stress response (1)
- integrin α2 (1)
- interleukin-6-type cytokines (1)
- internalization (1)
- interspecies comparison (1)
- intestine (1)
- intoxication (1)
- intracranial bleeding (1)
- intrahelical lesion (1)
- intrinsic metabolism (1)
- invasion (1)
- ion channels in the nervous system (1)
- ionic selectivity (1)
- iota-toxin (1)
- kidneys (1)
- kinase Syk (1)
- kinease (1)
- kinetics (1)
- knockout (1)
- laminin receptor (1)
- latency (1)
- learning (1)
- lesion formation (1)
- leukemia-inhibitory factor (1)
- leukocyte adhesion (1)
- ligand potencies (1)
- ligases (1)
- light pulses (1)
- light-induced interstrand DNA crosslinking (1)
- lightsheet microscopy (1)
- lines (1)
- lipid asymmetry (1)
- lipid bilayer membranes (1)
- lipidomics (1)
- listeriolysin O (1)
- liver (1)
- localization (1)
- locomotion (1)
- long-term depression (1)
- long-term potentation (1)
- loop B (1)
- loss of function (1)
- low-secretion phenotype mutants (1)
- lung and intrathoracic tumors (1)
- lymph node stromal cells (1)
- lymph node transplantation (1)
- lymph nodes (1)
- lymphocyte homing (1)
- lymphoma (1)
- lysosomal degradation (1)
- mRNA (1)
- magnetic resonance imaging (1)
- marcophages (1)
- mass spectrometry (1)
- mast cells (1)
- mastectomy (1)
- mastocytosis (1)
- mathematical modeling (1)
- matrix metalloproteinase (1)
- maturation signal (1)
- maus (1)
- mechanotransduction (1)
- medicine (1)
- medium spiny neurons (1)
- megakaryopoiesis (1)
- melanoma malignancy (1)
- melt electrowriting (1)
- membrane interaction (1)
- mesenteric lymph node (1)
- metabolism (1)
- metabotropic signalling (1)
- metalloproteinase (1)
- metals (1)
- metastases (1)
- miR-17~92 (1)
- miR-30 (1)
- miRNA (1)
- miRNA expression (1)
- miRNA processing (1)
- microenvironment (1)
- microglia (1)
- microscopy (1)
- microtubules (1)
- microvascular complications (1)
- microvascular density (1)
- microvasculature (1)
- midbody (1)
- migration (1)
- mismatch recognition (1)
- mismatches (1)
- mitochondria (1)
- mitochondrial transport (1)
- mitogenic signaling (1)
- mitosis (1)
- mobilization (1)
- model (1)
- modulation of virus replication (1)
- molecular basis (1)
- molecular mass (1)
- molecular mobility (1)
- molecular structure (1)
- molybdenum cofactor biosynthesis (1)
- monoclonal antibodies (1)
- monocyte (1)
- monocytes (1)
- monovalent cation:proton antiporter-2 (CPA2) family (1)
- morphogenesis (1)
- motion (1)
- motoneuron (1)
- movement disorders (1)
- mrsk2 KO mouse (1)
- multi-drug-resistance (1)
- multiphoton microscopy (1)
- multiple myeloma (1)
- multiple sclerosis (1)
- murine cytomegalovirus (1)
- muscle (1)
- mutant p53 (1)
- mutation screening (1)
- mutation triggers (1)
- mutations (1)
- mycobacterium smegmatis (1)
- mycolic acid (1)
- myogenic differentiation (1)
- myomesin (1)
- nanoelectronics (1)
- nanolithography (1)
- nanomedicine (1)
- nanorobot (1)
- nanotechnology (1)
- naphthylisoquinoline alkaloids (1)
- neddylation (1)
- neocortex (1)
- network (1)
- neural networks (1)
- neurite outgrowth (1)
- neuroblastoma cell (1)
- neurologin-2 (1)
- neurology (1)
- neuromuscular junction (1)
- neuronal differentiation (1)
- neuropil (1)
- neurotransmitters (1)
- neurotrophic factor (1)
- neutralization (1)
- neutrophil (1)
- neutrophils (1)
- nilotinib (1)
- nitric oxide synthase (1)
- non-apoptotic programmed cell death (1)
- non-canonical autophagy (1)
- noncooperative binding (1)
- nuclear envelope (1)
- nuclear export (1)
- nuclear receptors (1)
- obesity (1)
- object segmentation (1)
- obscurin (1)
- octopamine (1)
- oncolytic viral therapy (1)
- oncolytic virotherapy (1)
- opioid peptides (1)
- opioid receptors (1)
- outer membrane proteins (1)
- oxazolone colitis (1)
- oxidation (1)
- oxidative stress (1)
- p-cresyl sulfate (1)
- p53-dependent apoptosis (1)
- p53-inducible regulator (1)
- pain behavior (1)
- pancreas (1)
- parathyroid hormone 1 receptor (1)
- pathway analysis (1)
- peptide (1)
- peptide inhibitor design (1)
- peptide inhibitor of envelopment (1)
- peptide microarray (1)
- peptides (1)
- peptidomoics (1)
- peripheral nerve (1)
- peripheral nervous system (1)
- permeability (1)
- pet dogs (1)
- phagolysosome tubulation (1)
- phosphatidic acid (1)
- phosphatidylethanolamine (1)
- phosphatidylserine (1)
- phosphoglycolate phosphatase (1)
- phospholipase C gamma 2 (1)
- phosphorylation sites (1)
- photo-cross-linking (1)
- photoconversion (1)
- pkd (1)
- planar cell polarity (1)
- plasma membrane (1)
- plasticity (1)
- platelet adhesion (1)
- platelet degranulation (1)
- platelet factor 4 (1)
- platelet inhibition (1)
- platelet receptor (1)
- platelet-neutrophil complexes (PNCs) (1)
- pneumolysin (1)
- pocket factor (1)
- podoplanin (1)
- polar body (1)
- polarization (1)
- popliteal aneurysm (1)
- pore formation (1)
- pore-forming toxin (1)
- post-translational modification (1)
- post-translational modifications (1)
- potassium homeostasis (1)
- precursor cells (1)
- presynaptic hyperekplexia (1)
- processivity (1)
- progressive encephalitis with rigidity and myoclonus (PERM) (1)
- proliferating cell nuclear antigen (PCNA) (1)
- promoter affinity (1)
- propionic acid (1)
- proteasome inhibitor resistance (1)
- proteasome subunit beta type-5 (PSMB5) (1)
- proteasome system (1)
- protective antigen (1)
- protein NEDD8 (1)
- protein RSK2 (1)
- protein disulfide isomerase (1)
- protein folding (1)
- protein kinase (1)
- protein kinase C (1)
- protein kinase D1 (1)
- protein kinase D2/PKD2/PRKD2 (1)
- protein processing (1)
- protein structures (1)
- protein synthesis shut down (1)
- protein transport (1)
- protein ubiquitination (1)
- protein-DNA interaction (1)
- protein-DNA interactions (1)
- protein-protein interaction (1)
- protein-protein interaction (PPI) (1)
- proteins (1)
- protein–protein interaction (1)
- proteome (1)
- proteomics (1)
- protochlamydia amoebophila (1)
- protoplasts (1)
- pseudomonas aeruginosa (1)
- psoriasis (1)
- puberty (1)
- purification (1)
- pyridoxal phosphatase (1)
- pyridoxal phosphatase (PDXP) (1)
- radiation-therapy (1)
- radii (1)
- radioiodine therapy (1)
- rat spinal-cord (1)
- reaction mechanisms (1)
- receptor (1)
- receptor clustering (1)
- receptor signaling (1)
- receptor-beta subunits (1)
- recombinant tissue-type plasminogen activator (1)
- recruitment (1)
- regulation (1)
- regulatory T cells (1)
- regulatory circuit downstream (1)
- renal cancer (1)
- repair (1)
- repair and replication (1)
- replication (1)
- replication fork (1)
- resolvin (1)
- responses (1)
- restoration (1)
- restricition enzymes (1)
- rhabdomyosarcoma (1)
- rhabdomyosarcoma development (1)
- scaffold protein (1)
- scaffolding protein gephyryrin (1)
- screening (1)
- secondary lung tumors (1)
- secreted factors (1)
- selective channel (1)
- seminoma (1)
- senescence (1)
- sensitive amine oxidase (1)
- sensory cues (1)
- sensory physiology (1)
- serotonin (1)
- serum 25-hydroxyvitamin D (1)
- sex addiction (1)
- shedding (1)
- side chain properties (1)
- signal peptides (1)
- signal transduction (1)
- signal transduction pathway (1)
- signaling microdomain (1)
- signaling network (1)
- signalling pathways (1)
- silver staining (1)
- single channel analysis (1)
- single channel recording (1)
- single molecule (1)
- single strand blocking (1)
- site (1)
- size exclusion chromatography (1)
- skin autofluorescence (1)
- sleeping sickness (1)
- sliding (1)
- sodium-iodide symporter (1)
- sorafenib (1)
- spatiotemporal thrombus (1)
- species specificity (1)
- spinal-cord (1)
- spinco ultracentrifuge (1)
- splenic function (1)
- stage-i (1)
- startle (1)
- stem cells (1)
- stiff-person syndrome (SPS) (1)
- stomach (1)
- storage-pool diseases (1)
- store-operated calcium entry (1)
- stranded DNAs (1)
- stress response (1)
- structural biology (1)
- structural disruption (1)
- structural mechanism (1)
- structure-activity relationships (1)
- structure-based drug design (1)
- subcutaneous human tumors (1)
- subpopulation (1)
- subsets (1)
- substrate recognition (1)
- substrate specificity (1)
- subthreshold IKK activation (1)
- subunit (1)
- sulfates (1)
- sulfides (1)
- sulfites (1)
- super resolution microscopy (1)
- super-resolution microscopy (1)
- super-resolution microscopy (SRM) (1)
- surgery (1)
- swiss model (1)
- synapse (1)
- synapse formation (1)
- synaptic development (1)
- system (1)
- systemic micro-inflammation oxidative stress (1)
- tMCAO (1)
- telomerase (1)
- temporal range (1)
- temporal-lobe epilepsy (1)
- tetraspanin (1)
- thiol starvation (1)
- thiosulfates (1)
- thromboxane (1)
- thymine (1)
- thyroid-cancer (1)
- time-correlated single photon counting (TCSPC) (1)
- time-resolved anisotropy (1)
- titin (1)
- torque (1)
- torque meter (1)
- toxin (1)
- toxins (1)
- trafficking (1)
- trans-Golgi network (1)
- transactivation (1)
- transcription (1)
- transcription factors (1)
- transcriptome (1)
- transient middle cerebral artery occlusion (1)
- transient receptor potential channels (1)
- translesion synthesis (1)
- transmission (1)
- trypanosomes (1)
- tuberculosis (1)
- tumor microenvironment (1)
- tumour-necrosis factors (1)
- type 1 diabetes (1)
- type I interferon (1)
- tyrosine-protein kinase (1)
- ubiquitin ligase (1)
- ubiquitylation (ubiquitination) (1)
- ultrastructure (1)
- undamaged DNA (1)
- uracil binding (1)
- variocosities (1)
- vascular adhesion protein-1 (1)
- vascular system (1)
- vaskuläre glatte Muskelzelle (1)
- vasp (1)
- velocity (1)
- vibration (1)
- virotherapy (1)
- virus reactivation (1)
- walking (1)
- x-ray crystallography (1)
- xeroderma-pigmentosum (1)
- µ-Opioid receptor (1)
- Übergewicht (1)
- β cell (1)
- β1-adrenoceptor/β1-adrenergic receptor (1)
- β3 adrenergic receptor (1)
- β3 adrenergic receptor (ADRB3) (1)
Institute
- Rudolf-Virchow-Zentrum (305) (remove)
Sonstige beteiligte Institutionen
- Rudolf Virchow Center for Integrative and Translational Bioimaging, University of Würzburg (2)
- Center for Nanosystems Chemistry (CNC), University of Würzburg (1)
- Eberhard Karls Universität Tübingen (1)
- Genelux Corporation, San Diego Science Center, 3030 Bunker Hill Street, Suite 310, San Diego, California 92109, USA (1)
- MRB Forschungszentrum für Magnet-Resonanz-Bayern e.V., Am Hubland, D-97074 Würzburg (1)
- Research Center for Infectious Diseases, University of Wuerzburg, Wuerzburg 97080, Germany (1)
- Rudolf-Virchow-Zentrum für Experimentelle Biomedizin der Universität Würzburg (1)
Glycine receptor β–targeting autoantibodies contribute to the pathology of autoimmune diseases
(2024)
Background and Objectives
Stiff-person syndrome (SPS) and progressive encephalomyelitis with rigidity and myoclonus (PERM) are rare neurologic disorders of the CNS. Until now, exclusive GlyRα subunit–binding autoantibodies with subsequent changes in function and surface numbers were reported. GlyR autoantibodies have also been described in patients with focal epilepsy. Autoimmune reactivity against the GlyRβ subunits has not yet been shown. Autoantibodies against GlyRα1 target the large extracellular N-terminal domain. This domain shares a high degree of sequence homology with GlyRβ making it not unlikely that GlyRβ-specific autoantibody (aAb) exist and contribute to the disease pathology.
Methods
In this study, we investigated serum samples from 58 patients for aAb specifically detecting GlyRβ. Studies in microarray format, cell-based assays, and primary spinal cord neurons and spinal cord tissue immunohistochemistry were performed to determine specific GlyRβ binding and define aAb binding to distinct protein regions. Preadsorption approaches of aAbs using living cells and the purified extracellular receptor domain were further used. Finally, functional consequences for inhibitory neurotransmission upon GlyRβ aAb binding were resolved by whole-cell patch-clamp recordings.
Results
Among 58 samples investigated, cell-based assays, tissue analysis, and preadsorption approaches revealed 2 patients with high specificity for GlyRβ aAb. Quantitative protein cluster analysis demonstrated aAb binding to synaptic GlyRβ colocalized with the scaffold protein gephyrin independent of the presence of GlyRα1. At the functional level, binding of GlyRβ aAb from both patients to its target impair glycine efficacy.
Discussion
Our study establishes GlyRβ as novel target of aAb in patients with SPS/PERM. In contrast to exclusively GlyRα1-positive sera, which alter glycine potency, aAbs against GlyRβ impair receptor efficacy for the neurotransmitter glycine. Imaging and functional analyses showed that GlyRβ aAbs antagonize inhibitory neurotransmission by affecting receptor function rather than localization.
The cystine/glutamate antiporter xCT is an important source of cysteine for cancer cells. Once taken up, cystine is reduced to cysteine and serves as a building block for the synthesis of glutathione, which efficiently protects cells from oxidative damage and prevents ferroptosis. As melanomas are particularly exposed to several sources of oxidative stress, we investigated the biological role of cysteine and glutathione supply by xCT in melanoma. xCT activity was abolished by genetic depletion in the Tyr::CreER; Braf\(^{CA}\); Pten\(^{lox/+}\) melanoma model and by acute cystine withdrawal in melanoma cell lines. Both interventions profoundly impacted melanoma glutathione levels, but they were surprisingly well tolerated by murine melanomas in vivo and by most human melanoma cell lines in vitro. RNA sequencing of human melanoma cells revealed a strong adaptive upregulation of NRF2 and ATF4 pathways, which orchestrated the compensatory upregulation of genes involved in antioxidant defence and de novo cysteine biosynthesis. In addition, the joint activation of ATF4 and NRF2 triggered a phenotypic switch characterized by a reduction of differentiation genes and induction of pro-invasive features, which was also observed after erastin treatment or the inhibition of glutathione synthesis. NRF2 alone was capable of inducing the phenotypic switch in a transient manner. Together, our data show that cystine or glutathione levels regulate the phenotypic plasticity of melanoma cells by elevating ATF4 and NRF2.
Vitamin B6 deficiency has been linked to cognitive impairment in human brain disorders for decades. Still, the molecular mechanisms linking vitamin B6 to these pathologies remain poorly understood, and whether vitamin B6 supplementation improves cognition is unclear as well. Pyridoxal phosphatase (PDXP), an enzyme that controls levels of pyridoxal 5’-phosphate (PLP), the co-enzymatically active form of vitamin B6, may represent an alternative therapeutic entry point into vitamin B6-associated pathologies. However, pharmacological PDXP inhibitors to test this concept are lacking. We now identify a PDXP and age-dependent decline of PLP levels in the murine hippocampus that provides a rationale for the development of PDXP inhibitors. Using a combination of small molecule screening, protein crystallography and biolayer interferometry, we discover and analyze 7,8-dihydroxyflavone (7,8-DHF) as a direct and potent PDXP inhibitor. 7,8-DHF binds and reversibly inhibits PDXP with low micromolar affinity and sub-micromolar potency. In mouse hippocampal neurons, 7,8-DHF increases PLP in a PDXP-dependent manner. These findings validate PDXP as a druggable target. Of note, 7,8-DHF is a well-studied molecule in brain disorder models, although its mechanism of action is actively debated. Our discovery of 7,8-DHF as a PDXP inhibitor offers novel mechanistic insights into the controversy surrounding 7,8-DHF-mediated effects in the brain.
Besides their central role in haemostasis and thrombosis, platelets are increasingly recognised as versatile effector cells in inflammation, the innate and adaptive immune response, extracellular matrix reorganisation and fibrosis, maintenance of barrier and organ integrity, and host response to pathogens. These platelet functions, referred to as thrombo-inflammation and immunothrombosis, have gained major attention in the COVID-19 pandemic, where patients develop an inflammatory disease state with severe and life-threatening thromboembolic complications. In the CRC/TR 240, a highly interdisciplinary team of basic, translational and clinical scientists explored these emerging roles of platelets with the aim to develop novel treatment concepts for cardiovascular disorders and beyond. We have i) unravelled mechanisms leading to life-threatening thromboembolic complica-tions following vaccination against SARS-CoV-2 with adenoviral vector-based vaccines, ii) identified unrecognised functions of platelet receptors and their regulation, offering new potential targets for pharmacological intervention and iii) developed new methodology to study the biology of megakar-yocytes (MKs), the precursor cells of platelets in the bone marrow, which lay the foundation for the modulation of platelet biogenesis and function. The projects of the CRC/TR 240 built on the unique expertise of our research network and focussed on the following complementary fields: (A) Cell bi-ology of megakaryocytes and platelets and (B) Platelets as regulators and effectors in disease. To achieve this aim, we followed a comprehensive approach starting out from in vitro systems and animal models to clinical research with large prospective patient cohorts and data-/biobanking. Despite the comparably short funding period the CRC/TR 240 discovered basic new mechanisms of platelet biogenesis, signal transduction and effector function and identified potential MK/platelet-specific molecular targets for diagnosis and therapy of thrombotic, haemorrhagic and thrombo-inflammatory disease states.
Platelets play an important role in haemostasis by mediating blood clotting at sites of blood vessel damage. Platelets, also participate in pathological conditions including thrombosis and inflammation. Upon vessel damage, two glycoprotein receptors, the GPIb-IX-V complex and GPVI, play important roles in platelet capture and activation.
GPIb-IX-V binds to von Willebrand factor and GPVI to collagen. This initiates a signalling cascade resulting in platelet shape change and spreading, which is dependent on the actin cytoskeleton. This thesis aimed to develop and implement different super-resolution microscopy techniques to gain a deeper understanding of the conformation and location of these receptors in the platelet plasma membrane, and to provide insights into their signalling pathways. We suggest direct stochastic optical reconstruction microscopy (dSTORM) and structured illumination microscopy (SIM) as the best candidates for imaging single platelets, whereas expansion microscopy (ExM) is ideal for imaging platelets aggregates.
Furthermore, we highlighted the role of the actin cytoskeleton, through Rac in GPVI signalling pathway. Inhibition of Rac, with EHT1864 in human platelets induced GPVI and GPV, but not GPIbα shedding. Furthermore, EHT1864 treatment did not change GPVI dimerisation or clustering, however, it decreased phospholipase Cγ2 phosphorylation levels, in human, but not murine platelets, highlighting interspecies differences. In summary, this PhD thesis demonstrates that; 1) Rac alters GPVI signalling pathway in human but not mouse platelets; 2) our newly developed ExM protocol can be used to image platelet aggregates labelled with F(ab’) fragments
Sepsis ist ein häufiges und akut lebensbedrohliches Syndrom, das eine Organfunktionsstörung in Folge einer dysregulierten Immunantwort auf eine Infektion beschreibt. Eine frühzeitige Diagnosestellung und Therapieeinleitung sind von zentraler Bedeutung für das Überleben der Patient:innen. In einer Pilotstudie konnte unsere Forschungsgruppe mittels Durchflusszytometrie eine ausgeprägte Hyporeaktivität der Thrombozyten bei Sepsis nachweisen, die einen potenziell neuen Biomarker zur Sepsis-Früherkennung darstellt. Zur Evaluation des Ausmaßes und Entstehungszeitpunktes der detektierten Thrombozytenfunktionsstörung wurden im Rahmen der vorliegenden Arbeit zusätzlich zu Patient:innen mit Sepsis (SOFA-Score ≥ 2; n=13) auch hospitalisierte Patient:innen mit einer Infektion ohne Sepsis (SOFA-Score < 2; n=12) rekrutiert. Beide Kohorten wurden zu zwei Zeitpunkten (t1: <24h; t2: Tag 5-7) im Krankheitsverlauf mittels Durchflusszytometrie und PFA-200 untersucht und mit einer gesunden Kontrollgruppe (n=28) verglichen.
Phänotypische Auffälligkeiten der Thrombozyten bei Sepsis umfassten: (i) eine veränderte Expression verschiedener Untereinheiten des GPIb-IX-V-Rezeptorkomplexes, die auf ein verstärktes Rezeptor-Shedding hindeutet; (ii) ein ausgeprägtes Mepacrin-Beladungsdefizit, das auf eine zunehmend reduzierte Anzahl von δ-Granula entlang des Infektion-Sepsis Kontinuums hinweist; (iii) eine Reduktion endständig gebundener Sialinsäure im Sinne einer verstärkten Desialylierung. Die funktionelle Analyse der Thrombozyten bei Sepsis ergab bei durchflusszytometrischer Messung der Integrin αIIbβ3-Aktivierung (PAC-1-Bindung) eine ausgeprägte generalisierte Hyporeaktivität gegenüber multiplen Agonisten, die abgeschwächt bereits bei Infektion nachweisbar war und gemäß ROC-Analysen gut zwischen Infektion und Sepsis diskriminierte (AUC >0.80 für alle Agonisten). Im Gegensatz dazu zeigten Thrombozyten bei Sepsis und Analyse mittels PFA-200 unter Einfluss physiologischer Scherkräfte eine normale bis gar beschleunigte Aggregation.
Die Reaktivitätsmessung von Thrombozyten mittels Durchflusszytometrie stellt weiterhin einen vielversprechenden Biomarker für die Sepsis-Früherkennung dar. Für weitere Schlussfolgerungen ist jedoch eine größere Kohorte erforderlich. In nachfolgenden Untersuchungen sollten zudem mechanistische Ursachen der beschriebenen phänotypischen und funktionellen Auffälligkeiten von Thrombozyten bei Infektion und Sepsis z.B. mittels Koinkubationsexperimenten untersucht werden.
The pancreas is the key organ for the maintenance of euglycemia. This is regulated in particular by α-cell-derived glucagon and β-cell-derived insulin, which are released in response to nutrient deficiency and elevated glucose levels, respectively. Although glucose is the main regulator of insulin secretion, it is significantly enhanced by various potentiators.
Platelets are anucleate cell fragments in the bloodstream that are essential for hemostasis to prevent and stop bleeding events. Besides their classical role, platelets were implemented to be crucial for other physiological and pathophysiological processes, such as cancer progression, immune defense, and angiogenesis. Platelets from diabetic patients often present increased reactivity and basal activation. Interestingly, platelets store and release several substances that have been reported to potentiate insulin secretion by β-cells. For these reasons, the impact of platelets on β-cell functioning was investigated in this thesis.
Here it was shown that both glucose and a β-cell-derived substance/s promote platelet activation and binding to collagen. Additionally, platelet adhesion specifically to the microvasculature of pancreatic islets was revealed, supporting the hypothesis of their influence on glucose homeostasis. Genetic or pharmacological ablation of platelet functioning and platelet depletion consistently resulted in reduced insulin secretion and associated glucose intolerance. Further, the platelet-derived lipid fraction was found to enhance glucose-stimulated insulin secretion, with 20-hydroxyeicosatetraenoic acid (20-HETE) and possibly also lyso-precursor of platelet-activating factor (lysoPAF) being identified as crucial factors. However, the acute platelet-stimulated insulin secretion was found to decline with age, as did the levels of platelet-derived 20-HETE. In addition to their direct stimulatory effect on insulin secretion, specific defects in platelet activation have also been shown to affect glucose homeostasis by potentially influencing islet vascular development. Taking together, the results of this thesis suggest a direct and indirect mechanism of platelets in the regulation of insulin secretion that ensures glucose homeostasis, especially in young individuals.
Ubiquitination is an important post-translational modification that maintains cellular homeostasis by regulating various biological processes. Deubiquitinases (DUBs) are enzymes that reverse the ubiquitination process by catalyzing the removal of ubiquitin from a substrate. Abnormal expression or function of DUBs is often associated with the onset and progression of various diseases, including cancer. Ubiquitin specific proteases (USPs), which constitute the largest family of DUBs in humans, have become the center of interest as potential targets in cancer therapy as many of them display increased activity or are overexpressed in a range of malignant tumors or the tumor microenvironment.
Two related members of the USP family, USP28 and USP25, share high sequence identities but play diverse biological roles. USP28 regulates cell proliferation, oncogenesis, DNA damage repair and apoptosis, whereas USP25 is involved in the anti-viral response, innate immunity and ER-associated degradation in addition to carcinogenesis. USP28 and USP25 also exhibit different oligomeric states – while USP28 is a constitutively active dimer, USP25 assumes an auto-inhibited tetrameric structure. The catalytic domains of both USP28 and USP25 comprise the canonical, globular USP-domain but contain an additional, extended insertion site called USP25/28 catalytic domain inserted domain (UCID) that mediates oligomerization of the proteins. Disruption of the USP25 tetramer leads to the formation of an activated dimeric protein. However, it is still not clear what triggers its activation.
Due to their role in maintaining and stabilizing numerous oncoproteins, USP28 and USP25 have emerged as interesting candidates for anti-cancer therapy. Recent advances in small-molecular inhibitor development have led to the discovery of relatively potent inhibitors of USP28 and USP25. This thesis focuses on the structural elucidation of USP28 and the biochemical characterization of USP28/USP25, both in complex with representatives of three out of the eight compound classes reported as USP28/USP25-specific inhibitors. The crystal structures of USP28 in complex with the AZ compounds, Vismodegib and FT206 reveal that all three inhibitor classes bind into the same allosteric pocket distant from the catalytic center, located between the palm and the thumb subdomains (the S1-site). Intriguingly, this binding pocket is identical to the UCID-tip binding interface in the USP25 tetramer, rendering the protein in a locked, inactive conformation. Formation of the binding pocket in USP28 requires a shift in the helix α5, which induces conformational changes and local distortion of the binding channel that typically accommodates the C-terminal tail of Ubiquitin, thus preventing catalysis and abrogating USP28 activity. The key residues of the USP28-inhibitor binding pocket are highly conserved in USP25. Mutagenesis studies of these residues accompanied by biochemical and biophysical assays confirm the proposed mechanism of inhibition and similar binding to USP25.
This work provides valuable insights into the inhibition mechanism of the small molecule compounds specifically for the DUBs USP28 and USP25. The USP28-inhibitor complex structures offer a framework to develop more specific and potent inhibitors.
Stroke and myocardial infarction are the most prominent and severe consequences of pathological thrombus formation. For prevention and/or treatment of thrombotic events there is a variety of anti-coagulation and antiplatelet medication that all have one side effect in common: the increased risk of bleeding. To design drugs that only intervene in the unwanted aggregation process but do not disturb general hemostasis, it is crucial to decipher the exact clotting pathway which has not been fully understood yet. Platelet membrane receptors play a vital role in the clotting pathway and, thus, the aim of this work is to establish a method to elucidate the interactions, clustering, and reorganization of involved membrane receptors such as GPIIb/IIIa and GPIX as part of the GPIb-IX-V complex. The special challenges regarding visualizing membrane receptor interactions on blood platelets are the high abundancy of the first and the small size of the latter (1—3µm of diameter). The resolution limit of conventional fluorescence microscopy and even super-resolution approaches prevents the successful differentiation of densely packed receptors from one another. Here, this issue is approached with the combination of a recently developed technique called Expansion Microscopy (ExM). The image resolution of a conventional fluorescence microscope is enhanced by simply enlarging the sample physically and thus pulling the receptors apart from each other. This method requires a complex sample preparation and holds lots of obstacles such as variable or anisotropic expansion and low images contrast. To increase ExM accuracy and sensitivity for interrogating blood platelets, it needs optimized sample preparation as well as image analysis pipelines which are the main part of this thesis. The colocalization results show that either fourfold or tenfold expanded, resting platelets allow a clear distinction between dependent, clustered, and independent receptor organizations compared to unexpanded platelets.Combining dual-color Expansion and confocal fluorescence microscopy enables to image in the nanometer range identifying GPIIb/IIIa clustering in resting platelets – a pattern that may play a key role in the clotting pathway
This decade saw the development of new high-end light microscopy approaches. These technologies are increasingly used to expand our understanding of cellular function and the molecular mechanisms of life and disease. The precision of state-of-the-art super resolution microscopy is limited by the properties of the applied fluorescent label. Here I describe the synthesis and evaluation of new functional fluorescent probes that specifically stain gephyrin, universal marker of the neuronal inhibitory post-synapse. Selected probe precursor peptides were synthesised using solid phase peptide synthesis and conjugated with selected super resolution capable fluorescent dyes. Identity and purity were defined using chromatography and mass spectrometric methods. To probe the target specificity of the resulting probe variants in cellular context, a high-throughput assay was established. The established semi-automated and parallel workflow was used for the evaluation of three selected probes by defining their co-localization with the expressed fluorescent target protein. My work provided NN1Dc and established the probe as a visualisation tool for essentially background-free visualisation of the synaptic marker protein gephyrin in a cellular context. Furthermore, NN1DA became part of a toolbox for studying the inhibitory synapse ultrastructure and brain connectivity and turned out useful for the development of a label-free, high-throughput protein interaction quantification assay.
Formation of the Aurora-A–MYCN complex increases levels of the oncogenic transcription factor MYCN in neuroblastoma cells by abrogating its degradation through the ubiquitin proteasome system. While some small-molecule inhibitors of Aurora-A were shown to destabilize MYCN, clinical trials have not been satisfactory to date. MYCN itself is considered to be `undruggable' due to its large intrinsically disordered regions. Targeting the Aurora-A–MYCN complex rather than Aurora-A or MYCN alone will open new possibilities for drug development and screening campaigns. To overcome the challenges that a ternary system composed of Aurora-A, MYCN and a small molecule entails, a covalently cross-linked construct of the Aurora-A–MYCN complex was designed, expressed and characterized, thus enabling screening and design campaigns to identify selective binders.
Covalent crosslinking of DNA strands provides a useful tool for medical, biochemical and DNA nanotechnology applications. Here we present a light-induced interstrand DNA crosslinking reaction using the modified nucleoside 5-phenylethynyl-2’-deoxyuridine (\(^{Phe}\)dU). The crosslinking ability of \(^{Phe}\)dU was programmed by base pairing and by metal ion interaction at the Watson-Crick base pairing site. Rotation to intrahelical positions was favored by hydrophobic stacking and enabled an unexpected photochemical alkene-alkyne [2+2] cycloaddition within the DNA duplex, resulting in efficient formation of a \(^{Phe}\)dU-dimer after short irradiation times of a few seconds. A \(^{Phe}\)dU dimer-containing DNA was shown to efficiently bind a helicase complex, but the covalent crosslink completely prevented DNA unwinding, suggesting possible applications in biochemistry or structural biology.
Platelets have a key physiological role in haemostasis however, inappropriate thrombus formation can lead to cardiovascular diseases such as myocardial infarction or stroke. Although, such diseases are common worldwide there are comparatively few anti-platelet drugs, and these are associated with an increased risk of bleeding. Platelets also have roles in thrombo-inflammation, immuno-thrombosis and cancer, in part via C-type lectin-like receptor 2 (CLEC-2) and its ligand podoplanin. Although CLEC-2 contributes to these diseases in mice, as well as to thrombus stability, it is unclear whether CLEC-2 has similar roles in humans, particularly as human CLEC-2 (hCLEC-2) cannot be investigated experimentally in vivo.
To investigate hCLEC-2 in vivo, we generated a humanised CLEC-2 mouse (hCLEC-2KI) model, as well as a novel monoclonal antibody, HEL1, that binds to a different site than an existing antibody, AYP1. Using these antibodies, we have provided proof of principle for the use of hCLEC-2KI mice to test potential therapeutics targeting hCLEC-2, and shown for the first time that hCLEC-2 can be immunodepleted, with little effect on haemostasis. However, our results have also suggested that there are species differences in the role of CLEC-2 in arterial thrombosis. We further confirmed this using human blood where blocking CLEC-2 ligand binding had no effect on thrombosis, whereas we confirmed a minor role for mouse CLEC-2 in thrombus stability. We also investigated the effect of blocking CLEC-2 signalling using the Bruton’s tyrosine kinase inhibitor PRN473 on CLEC-2 mediated immuno-thrombosis in a Salmonella typhimurium infection model. However, no effect on thrombosis was observed suggesting that CLEC-2 signalling is not involved.
Overall, our results suggest that there may be differences in the role of human and mouse CLEC-2, at least in arterial thrombosis, which could limit the potential of CLEC-2 as an anti-thrombotic target. However, it appears that the interaction between CLEC-2 and podoplanin is conserved and therefore CLEC-2 could still be a therapeutic target in immuno-thrombosis, thrombo-inflammation and cancer. Furthermore, any potential human specific therapeutics could be investigated in vivo using hCLEC-2KI mice.
Kef couples the potassium efflux with proton influx in gram-negative bacteria. The resulting acidification of the cytosol efficiently prevents the killing of the bacteria by reactive electrophilic compounds. While other degradation pathways for electrophiles exist, Kef is a short-term response that is crucial for survival. It requires tight regulation since its activation comes with the burden of disturbed homeostasis. Electrophiles, entering the cell, react spontaneously or catalytically with glutathione, which is present at high concentrations in the cytosol. The resulting glutathione conjugates bind to the cytosolic regulatory domain of Kef and trigger activation while the binding of glutathione keeps the system closed. Furthermore, nucleotides can bind to this domain for stabilization or inhibition. The binding of an additional ancillary subunit, called KefF or KefG, to the cytosolic domain is required for full activation. The regulatory domain is termed K+ transport–nucleotide binding (KTN) or regulator of potassium conductance (RCK) domain, and it is also found in potassium uptake systems or channels in other oligomeric arrangements. Bacterial RosB-like transporters and K+ efflux antiporters (KEA) of plants are homologs of Kef but fulfill different functions. In summary, Kef provides an interesting and well-studied example of a highly regulated bacterial transport system.
Short functional peptidic probes can maximize the potential of high-end microscopy techniques and multiplex imaging assays and provide new insights into normal and aberrant molecular, cellular and tissue function. Particularly, the visualization of inhibitory synapses requires protocol tailoring for different sample types and imaging techniques and relies either on genetic manipulation or on antibodies that underperform in tissue immunofluorescence. Starting from an endogenous activity-related ligand of gephyrin, a universal marker of the inhibitory post-synapse, I developed a short peptidic multivalent binder with exceptional affinity and selectivity to gephyrin. By tailoring fluorophores to the binder, I have obtained Sylite, a probe for the visualization of inhibitory synapses, with an outstanding signal-to-background ratio, that bests the “gold standard” gephyrin antibodies both in selectivity and in tissue immunofluorescence. In tissue Sylite benefits from simplified handling, provides robust synaptic labeling in record-short time and, unlike antibodies, is not affected by staining artefacts. In super-resolution microscopy Sylite precisely localizes the post-synapse and enables accurate pre- to post-synapse measurements. Combined with complimentary tracing techniques Sylite reveals inhibitory connectivity and profiles inhibitory inputs and synapse sizes of excitatory and inhibitory neurons in the periaqueductal gray brain region. Lastly, upon probe optimization for live cell application and with the help of novel thiol-reactive cell penetrating peptide I have visualized inhibitory synapses in living neurons. Taken together, my work provided a versatile probe for conventional and super-resolution microscopy and a workflow for the development and application of similar compact functional synthetic probes.
Introduction
Pro-thrombotic events are one of the prevalent causes of intensive care unit (ICU) admissions among COVID-19 patients, although the signaling events in the stimulated platelets are still unclear.
Methods
We conducted a comparative analysis of platelet transcriptome data from healthy donors, ICU, and non-ICU COVID-19 patients to elucidate these mechanisms. To surpass previous analyses, we constructed models of involved networks and control cascades by integrating a global human signaling network with transcriptome data. We investigated the control of platelet hyperactivation and the specific proteins involved.
Results
Our study revealed that control of the platelet network in ICU patients is significantly higher than in non-ICU patients. Non-ICU patients require control over fewer proteins for managing platelet hyperactivity compared to ICU patients. Identification of indispensable proteins highlighted key subnetworks, that are targetable for system control in COVID-19-related platelet hyperactivity. We scrutinized FDA-approved drugs targeting indispensable proteins and identified fostamatinib as a potent candidate for preventing thrombosis in COVID-19 patients.
Discussion
Our findings shed light on how SARS-CoV-2 efficiently affects host platelets by targeting indispensable and critical proteins involved in the control of platelet activity. We evaluated several drugs for specific control of platelet hyperactivity in ICU patients suffering from platelet hyperactivation. The focus of our approach is repurposing existing drugs for optimal control over the signaling network responsible for platelet hyperactivity in COVID-19 patients. Our study offers specific pharmacological recommendations, with drug prioritization tailored to the distinct network states observed in each patient condition. Interactive networks and detailed results can be accessed at https://fostamatinib.bioinfo-wuerz.eu/.
Glycine receptor (GlyR) autoantibodies are associated with stiff-person syndrome and the life-threatening progressive encephalomyelitis with rigidity and myoclonus in children and adults. Patient histories show variability in symptoms and responses to therapeutic treatments. A better understanding of the autoantibody pathology is required to develop improved therapeutic strategies. So far, the underlying molecular pathomechanisms include enhanced receptor internalization and direct receptor blocking altering GlyR function. A common epitope of autoantibodies against the GlyRα1 has been previously defined to residues 1A-33G at the N-terminus of the mature GlyR extracellular domain. However, if other autoantibody binding sites exist or additional GlyR residues are involved in autoantibody binding is yet unknown. The present study investigates the importance of receptor glycosylation for binding of anti-GlyR autoantibodies. The glycine receptor α1 harbors only one glycosylation site at the amino acid residue asparagine 38 localized in close vicinity to the identified common autoantibody epitope. First, non-glycosylated GlyRs were characterized using protein biochemical approaches as well as electrophysiological recordings and molecular modeling. Molecular modeling of non-glycosylated GlyRα1 did not show major structural alterations. Moreover, non-glycosylation of the GlyRα1N38Q did not prevent the receptor from surface expression. At the functional level, the non-glycosylated GlyR demonstrated reduced glycine potency, but patient GlyR autoantibodies still bound to the surface-expressed non-glycosylated receptor protein in living cells. Efficient adsorption of GlyR autoantibodies from patient samples was possible by binding to native glycosylated and non-glycosylated GlyRα1 expressed in living not fixed transfected HEK293 cells. Binding of patient-derived GlyR autoantibodies to the non-glycosylated GlyRα1 offered the possibility to use purified non-glycosylated GlyR extracellular domain constructs coated on ELISA plates and use them as a fast screening readout for the presence of GlyR autoantibodies in patient serum samples. Following successful adsorption of patient autoantibodies by GlyR ECDs, binding to primary motoneurons and transfected cells was absent. Our results indicate that the glycine receptor autoantibody binding is independent of the receptor’s glycosylation state. Purified non-glycosylated receptor domains harbouring the autoantibody epitope thus provide, an additional reliable experimental tool besides binding to native receptors in cell-based assays for detection of autoantibody presence in patient sera.
Ubiquitylation is a protein post translational modification, in which ubiquitin is covalently attached to target protein substrates resulting in diverse cellular outcomes. Besides ubiquitin, various ubiquitin-like proteins including FAT10 exist, which are also conjugated to target proteins. The underlying modification mechanisms are conserved. In the initial step, ubiquitin or a ubiquitin-like protein is thioester-linked to a catalytic cysteine in the E1activating enzyme in an ATP-dependent manner. The respective protein modifier is then transferred to an E2 conjugating enzyme in a transthioesterification reaction. Finally, an E3 ubiquitin ligase E3 catalyzes the covalent attachment of the protein modifier to a substrate. In the case of ubiquitin, multiple ubiquitin molecules can be attached to a substrate in the form of either linear or branched polyubiquitin chains but also as single ubiquitin modifications. Depending on the nature of the ubiquitin chain, the substrates are destined to various cellular processes such as their targeted destruction by the proteasome but also non-degradative outcomes may occur.
As stated above FAT10 is a ubiquitin-like protein modifier which typically targets proteins for proteasomal degradation. It consists of two ubiquitin-like domains and is mainly expressed in cells of the human immune system. The reported involvement of FAT10 modifications in cancers and other diseases has caught the attention of the scientific community as an inhibition of the FAT10ylation process may provide avenues for novel therapeutic approaches. UBA6 is the E1 activating enzyme that resides at the apex of the FAT10 proteasomal degradation pathway. UBA6 not only recognizes FAT10 but can also activate ubiquitin as efficiently as the ubiquitin specific E1 UBA1. The dual specificity of UBA6 may complicate the inhibition FAT10ylation since targeting the active site of UBA6 will also inhibit the UBA6-catalyzed ubiquitin activation. Therefore, it is important to understand the underlying principles for the dual specificity of UBA6 prior to the development of compounds interfering with FAT10ylation.
In this thesis important novel insights into the structure and function of UBA6 were derived by X-ray crystallography and biochemical methods. The first crystal structure of UBA6 reveals the multidomain architecture of this enzyme in atomic detail. The enzyme is composed of a rigid core including its active and inactive adenylation domains as well as a 4 helix bundle. Overall, the molecule adopts a “Y” shape architecture with the core at the base and the first and second catalytic half domains forming one arm of the “Y” and the ubiquitin fold domain constituting the other arm. While UBA6 shares the same domain architecture as UBA1, substantial differences were revealed by the crystal structure. In particular, the first catalytic half domain undergoes a significant shift to a position more distal from the core. This rigid body movement is assumed to generate room to accommodate the second ubiquitin-like domain of FAT10. Differences are also observed in a hydrophobic platform between the core and the first catalytic half domain and the adenylation active site in the core, which together from the binding sites for ubiquitin and FAT10. Site directed mutagenesis of key residues in these areas altered the UBA6-catalyzed activation of ubiquitin and FAT10. UBA6 variants were generated with the goal of trying to block the activation of FAT10 while still maintaining that of ubiquitin activation, in order to fully explain the dual specificity of UBA6. However, none of these mutations could block the activation of FAT10, while some of these UBA6 variants blocked ubiquitin activation. Preliminary inhibition assays with a group of E1 inhibitors belonging to the adenosyl sulfamate family demonstrated potent inhibition of FAT10ylation for two compounds. The dual specificity of UBA6 hence needs to be further examined by biochemical and structural methods. In particular, the structure of a complex between UBA6 and ubiquitin or FAT10 would provide key insights for further biochemical studies, ultimately allowing the targeted inhibition of the FAT10ylation machinery.
Monoglyceride lipase (MGL) hydrolyzes monoacylglycerols (MG) to glycerol and one fatty acid. Among the various MG species, MGL also degrades 2-arachidonoylglycerol, the most abundant endocannabinoid and potent activator of the cannabinoid receptors 1 and 2. We investigated the consequences of MGL deficiency on platelet function using systemic (Mgl\(^{−/−}\)) and platelet-specific Mgl-deficient (platMgl\(^{−/−}\)) mice. Despite comparable platelet morphology, loss of MGL was associated with decreased platelet aggregation and reduced response to collagen activation. This was reflected by reduced thrombus formation in vitro, accompanied by a longer bleeding time and a higher blood volume loss. Occlusion time after FeCl\(_3\)-induced injury was markedly reduced in Mgl\(^{−/−}\) mice, which is consistent with contraction of large aggregates and fewer small aggregates in vitro. The absence of any functional changes in platelets from platMgl\(^{−/−}\) mice is in accordance with lipid degradation products or other molecules in the circulation, rather than platelet-specific effects, being responsible for the observed alterations in Mgl\(^{−/−}\) mice. We conclude that genetic deletion of MGL is associated with altered thrombogenesis.