Refine
Has Fulltext
- yes (294)
Is part of the Bibliography
- yes (294)
Year of publication
Document Type
- Journal article (185)
- 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 microscopy (3)
- lipid bilayer (3)
- macrophages (3)
- mechanisms (3)
- protein (3)
- thrombin (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)
- 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)
- 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)
- extracellular domain (2)
- factor XII (2)
- fluorescence imaging (2)
- fluorescence resonance energy transfer (FRET) (2)
- gene expression (2)
- glycine receptor (2)
- gp130 (2)
- human (2)
- hydrodynamics (2)
- inflammation (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)
- 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)
- 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)
- tumorigenesis (2)
- tumors (2)
- type VII secretion system (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)
- 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-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)
- MYC (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 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)
- STIM2 (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 (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 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)
- 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 discovery (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)
- 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)
- enzymes (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)
- 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)
- 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)
- 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)
- neuroinflammation (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)
- 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)
- phagosome maturation (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)
- 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)
- 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)
- 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)
- 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-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 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 (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 (294) (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)
Ein sehr wichtiger Tumorsuppressor ist der Transkriptionsfaktor p53, der Zellschicksals-Entscheidungen wie Zellzyklus-Arrest und programmierten Zelltod (Apoptose) kontrolliert. Die Wirkung von p53 und von seinen Familienmitgliedern p63 und p73 beruht überwiegend auf der Fähigkeit, als Transkriptionsfaktoren die Genexpression zu regulieren. Die DNA-Bindung an Promotoren von Zielgenen ist dabei von grundlegender Bedeutung und wird durch die hoch konservierte zentrale DNA-Bindungs-Domäne und den Carboxy-Terminus bestimmt. In dieser Arbeit wurden die DNA-Bindungseigenschaften von p53 und verschiedener Carboxy-terminalen p73 Isoformen untersucht. In „electrophoretic mobility shift assay” (EMSA) Experimenten bildeten p53 und p73gamma nur schwache Sequenz-spezifische DNA-Komplexe, wohingegen p73alpha, beta und delta die DNA deutlich stärker banden. Die schwache DNA-Bindung von p53 und p73gamma kann durch mehrfach positiv geladene Carboxy-Termini erklärt werden, die über eine Sequenz-unabhängige DNA-Bindung ein Gleiten entlang der DNA ermöglichen. Die Deletion der Carboxy-terminalen Domäne (CTD) von p53 („p53delta30“) verstärkte dementsprechend die Sequenz-spezifische DNA-Bindung in vitro und seine Übertragung auf p73alpha („p73alpha+30“) schwächte sie ab. Mittels „fluorescence recovery after photobleaching“ (FRAP) Experimenten konnte in lebenden Zellen eine Verminderung der intra-nukleären Mobilität von p53 und p73alpha+30 durch die CTD gezeigt werden, die aus der Sequenz-unabhängigen DNA-Bindung resultiert. Zusätzlich reduzierte die CTD die Sequenz-spezifische DNA-Bindung von p53 an den p21 (CDKN1A) Promotor. Das Spektrum der regulierten Zielgene wurde in einer Genom-weiten Genexpressions-Analyse nicht durch die CTD verändert, sondern maßgeblich durch das Protein-Rückgrat von p53 beziehungsweise p73 bestimmt. Allerdings verminderte die CTD das Ausmaß der Transkriptions-Regulation und hemmte die Induktion von Zellzyklus-Arrest und Apoptose. Die mehrfach positiv geladene CTD in p53 besitzt demzufolge eine negativ regulatorische Wirkung, die in den wichtigsten p73 Isoformen alpha, beta und delta fehlt. Die zentrale DNA-Bindungs-Domäne trägt durch elektrostatische Wechselwirkungen zwischen H1-Helices (Aminosäurereste 177 bis 182) unterschiedlicher p53 Monomere zu kooperativer DNA-Bindung und zu Zellschicksals-Entscheidungen bei. Anhand von Mutanten, die unterschiedlich starke H1-Helix-Interaktionen ermöglichen, konnte gezeigt werden, dass starke Interaktionen die Bindung an Promotoren von pro-apoptotischen Genen verstärkte, wohingegen die Bindung an anti-apoptotische und Zellzyklus-blockierende Gene unabhängig von der Interaktions-Stärke war. Diese Unterschiede in der Promotor-Bindung ließen sich nicht auf eine veränderte zelluläre Lokalisation der Mutanten zurückführen, da alle Mutanten überwiegend nukleär lokalisiert waren. Eine an Serin 183 Phosphorylierungs-defekte Mutante von p53 bildete stabile DNA-Komplexe, entsprechend einer Mutante mit starker H1-Helix-Interaktion, und trans-aktivierte pro-apoptotische Promotoren stärker als Mutanten, die Phosphorylierung von p53 an Serin 183 simulieren. Da zusätzlich bekannt ist, dass Serin 183 mit der H1-Helix wechselwirkt, könnte diese Phosphorylierung einen physiologischen Mechanismus zur Regulation der H1-Helix-Interaktion und damit des Zellschicksals darstellen. Zusammenfassend ließ sich zeigen, dass sowohl die Interaktions-Stärke zweier DNA-Bindungs-Domänen als auch die elektrische Ladung des Carboxy-Terminus die DNA-Bindungseigenschaften von p53 Familienmitgliedern bestimmen und so Zellschicksals-Entscheidungen der p53 Familie beeinflussen.
Ligand-binding of Cys-loop receptors is determined by N-terminal extracellular loop structures from the plus as well as from the minus side of two adjacent subunits in the pentameric receptor complex. An aromatic residue in loop B of the glycine receptor (GIyR) undergoes direct interaction with the incoming ligand via a cation-π interaction. Recently, we showed that mutated residues in loop B identified from human patients suffering from hyperekplexia disturb ligand-binding. Here, we exchanged the affected human residues by amino acids found in related members of the Cys-loop receptor family to determine the effects of side chain volume for ion channel properties. GIyR variants were characterized in vitro following transfection into cell lines in order to analyze protein expression, trafficking, degradation and ion channel function. GIyR α1 G160 mutations significantly decrease glycine potency arguing for a positional effect on neighboring aromatic residues and consequently glycine-binding within the ligand-binding pocket. Disturbed glycinergic inhibition due to T162 α1 mutations is an additive effect of affected biogenesis and structural changes within the ligand-binding site. Protein trafficking from the ER toward the ER-Golgi intermediate compartment, the secretory Golgi pathways and finally the cell surface is largely diminished, but still sufficient to deliver ion channels that are functional at least at high glycine concentrations. The majority of T162 mutant protein accumulates in the ER and is delivered to ER-associated proteasomal degradation. Hence, G160 is an important determinant during glycine binding. In contrast, 1162 affects primarily receptor biogenesis whereas exchanges in functionality are secondary effects thereof.
Cholesterol-dependent cytolysins (CDCs) are protein toxins that originate from Gram-positive bacteria and contribute substantially to their pathogenicity. CDCs bind membrane cholesterol and build prepores and lytic pores. Some effects of the toxins are observed in non-lytic concentrations. Two pathogens, \(Streptococcus\) \(pneumoniae\) and \(Listeria\) \(monocytogenes\), cause fatal bacterial meningitis, and both produce toxins of the CDC family—pneumolysin and listeriolysin O, respectively. It has been demonstrated that pneumolysin produces dendritic varicosities (dendrite swellings) and dendritic spine collapse in the mouse neocortex, followed by synaptic loss and astrocyte cell shape remodeling without elevated cell death. We utilized primary glial cultures and acute mouse brain slices to examine the neuropathological effects of listeriolysin O and to compare it to pneumolysin with identical hemolytic activity. In cultures, listeriolysin O permeabilized cells slower than pneumolysin did but still initiated non-lytic astrocytic cell shape changes, just as pneumolysin did. In an acute brain slice culture system, listeriolysin O produced dendritic varicosities in an NMDA-dependent manner but failed to cause dendritic spine collapse and cortical astrocyte reorganization. Thus, listeriolysin O demonstrated slower cell permeabilization and milder glial cell remodeling ability than did pneumolysin and lacked dendritic spine collapse capacity but exhibited equivalent dendritic pathology.
We used a novel approach of cytostatically induced leucocyte depletion and subsequent reconstitution with leucocytes deprived of classical \((inflammatory/Gr1^{hi})\) or non-classical \((resident/Gr1^{lo})\) monocytes to dissect their differential role in atheroprogression under high-fat diet (HFD). Apolipoprotein E-deficient \((Apoe^{-/-})\) mice lacking classical but not non-classical monocytes displayed reduced lesion size and macrophage and apoptotic cell content. Conversely, HFD induced a selective expansion of classical monocytes in blood and bone marrow. Increased CXCL1 levels accompanied by higher expression of its receptor CXCR2 on classical monocytes and inhibition of monocytosis by CXCL1-neutralization indicated a preferential role for the CXCL1/CXCR2 axis in mobilizing classical monocytes during hypercholesterolemia. Studies correlating circulating and lesional classical monocytes in gene-deficient \(Apoe^{-/-}\) mice, adoptive transfer of gene-deficient cells and pharmacological modulation during intravital microscopy of the carotid artery revealed a crucial function of CCR1 and CCR5 but not CCR2 or \(CX_3CR1\) in classical monocyte recruitment to atherosclerotic vessels. Collectively, these data establish the impact of classical monocytes on atheroprogression, identify a sequential role of CXCL1 in their mobilization and CCR1/CCR5 in their recruitment.
Dishevelled (DVL) is the key component of the Wnt signaling pathway. Currently, DVL conformational dynamics under native conditions is unknown. To overcome this limitation, we develop the Fluorescein Arsenical Hairpin Binder- (FlAsH-) based FRET in vivo approach to study DVL conformation in living cells. Using this single-cell FRET approach, we demonstrate that (i) Wnt ligands induce open DVL conformation, (ii) DVL variants that are predominantly open, show more even subcellular localization and more efficient membrane recruitment by Frizzled (FZD) and (iii) Casein kinase 1 ɛ (CK1ɛ) has a key regulatory function in DVL conformational dynamics. In silico modeling and in vitro biophysical methods explain how CK1ɛ-specific phosphorylation events control DVL conformations via modulation of the PDZ domain and its interaction with DVL C-terminus. In summary, our study describes an experimental tool for DVL conformational sampling in living cells and elucidates the essential regulatory role of CK1ɛ in DVL conformational dynamics.
The eukaryotic actin cytoskeleton is an evolutionarily well-established pathogen target, as a large number of bacterial factors disturb its dynamics to alter the function of the host cells. These pathogenic factors modulate or mimic actin effector proteins or they modify actin directly, leading to an imbalance of the precisely regulated actin turnover. Here, we show that the pore-forming, cholesterol-dependent cytolysin pneumolysin (PLY), a major neurotoxin of Streptococcus pneumoniae, has the capacity to bind actin directly and to enhance actin polymerisation in vitro. In cells, the toxin co-localised with F-actin shortly after exposure, and this direct interaction was verified by Förster resonance energy transfer. PLY was capable of exerting its effect on actin through the lipid bilayer of giant unilamellar vesicles, but only when its pore competence was preserved. The dissociation constant of G-actin binding to PLY in a biochemical environment was 170–190 nM, which is indicative of a high-affinity interaction, comparable to the affinity of other intracellular actin-binding factors. Our results demonstrate the first example of a direct interaction of a pore-forming toxin with cytoskeletal components, suggesting that the cross talk between pore-forming cytolysins and cells is more complex than previously thought.
Lyme disease Borreliae are highly dependent on the uptake of nutrients provided by their hosts. Our study describes the identification of a 36 kDa protein that functions as putative dicarboxylate-specific porin in the outer membrane of Lyme disease Borrelia. The protein was purified by hydroxyapatite chromatography from Borrelia burgdorferi B31 and designated as DipA, for dicarboxylate-specific porin A. DipA was partially sequenced, and corresponding genes were identified in the genomes of B. burgdorferi B31, Borrelia garinii PBi and Borrelia afzelii PKo. DipA exhibits high homology to the Oms38 porins of relapsing fever Borreliae. B. burgdorferi DipA was characterized using the black lipid bilayer assay. The protein has a singlechannel conductance of 50 pS in 1 M KCl, is slightly selective for anions with a permeability ratio for cations over anions of 0.57 in KCl and is not voltage-dependent. The channel could be partly blocked by different di- and tricarboxylic anions. Particular high stability constants up to about 28,000 l/mol (in 0.1 M KCl) were obtained among the 11 tested anions for oxaloacetate, 2-oxoglutarate and citrate. The results imply that DipA forms a porin specific for dicarboxylates which may play an important role for the uptake of specific nutrients in different Borrelia species.
Xiphophorus fish exhibit a clear phenotypic polymorphism in puberty onset and reproductive strategies of males. In X. nigrensis and X. multilineatus, puberty onset is genetically determined and linked to a melanocortin 4 receptor (Mc4r) polymorphism of wild-type and mutant alleles on the sex chromosomes. We hypothesized that Mc4r mutant alleles act on wild-type alleles by a dominant negative effect through receptor dimerization, leading to differential intracellular signaling and effector gene activation. Depending on signaling strength, the onset of puberty either occurs early or is delayed. Here, we show by Förster Resonance Energy Transfer (FRET) that wild-type Xiphophorus Mc4r monomers can form homodimers, but also heterodimers with mutant receptors resulting in compromised signaling which explains the reduced Mc4r signaling in large males. Thus, hetero- vs. homo- dimerization seems to be the key molecular mechanism for the polymorphism in puberty onset and body size in male fish.
Interneuronal synaptic transmission relies on the proper spatial organization of presynaptic neurotransmitter release and its reception on the postsynaptic side by cognate neurotransmitter receptors. Neurotransmitter receptors are incorporated into and arranged within the plasma membrane with the assistance of scaffolding and adaptor proteins. At inhibitory GABAergic postsynapses, collybistin, a neuronal adaptor protein, recruits the scaffolding protein gephyrin and interacts with various neuronal factors including cell adhesion proteins of the neuroligin family, the GABAA receptor α2-subunit and the closely related small GTPases Cdc42 and TC10 (RhoQ). Most collybistin splice variants harbor an N-terminal SH3 domain and exist in an autoinhibited/closed state. Cdc42 and TC10, despite sharing 67.4% amino acid sequence identity, interact differently with collybistin. Here, we delineate the molecular basis of the collybistin conformational activation induced by TC10 with the aid of recently developed collybistin FRET sensors. Time-resolved fluorescence-based FRET measurements reveal that TC10 binds to closed/inactive collybistin leading to relief of its autoinhibition, contrary to Cdc42, which only interacts with collybistin when forced into an open state by the introduction of mutations destabilizing the closed state of collybistin. Taken together, our data describe a TC10-driven signaling mechanism in which collybistin switches from its autoinhibited closed state to an open/active state.
Enhanced expression of the MYC transcription factor is observed in the majority of tumors. Two seemingly conflicting models have been proposed for its function: one proposes that MYC enhances expression of all genes, while the other model suggests gene-specific regulation. Here, we have explored the hypothesis that specific gene expression profiles arise since promoters differ in affinity for MYC and high-affinity promoters are fully occupied by physiological levels of MYC. We determined cellular MYC levels and used RNA- and ChIP-sequencing to correlate promoter occupancy with gene expression at different concentrations of MYC. Mathematical modeling showed that binding affinities for interactions of MYC with DNA and with core promoter-bound factors, such as WDR5, are sufficient to explain promoter occupancies observed in vivo. Importantly, promoter affinity stratifies different biological processes that are regulated by MYC, explaining why tumor-specific MYC levels induce specific gene expression programs and alter defined biological properties of cells.
Fast inhibitory synaptic transmission is mediated by γ-aminobutyric acid type A receptors (GABAARs) that are enriched at functionally diverse synapses via mechanisms that remain unclear. Using isothermal titration calorimetry and complementary methods we demonstrate an exclusive low micromolar binding of collybistin to the α2-subunit of GABAARs. To explore the biological relevance of collybistin-α2-subunit selectivity, we generate mice with a mutation in the α2-subunit-collybistin binding region (Gabra2-1). The mutation results in loss of a distinct subset of inhibitory synapses and decreased amplitude of inhibitory synaptic currents. Gabra2–1 mice have a striking phenotype characterized by increased susceptibility to seizures and early mortality. Surviving Gabra2-1 mice show anxiety and elevations in electroencephalogram δ power, which are ameliorated by treatment with the α2/α3-selective positive modulator, AZD7325. Taken together, our results demonstrate an α2-subunit selective binding of collybistin, which plays a key role in patterned brain activity, particularly during development.
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
Posttranslational modifications (PTMs) play a crucial role in many cellular processes. They are reversible, dynamic, and highly regulated events that alter the properties of proteins and increase their functional diversity. The identification and quantification of PTMs are critical for deciphering the molecular mechanisms of PTMs-related biological processes and disease treatment and prevention. Two of the most common and important PTMs that regulate many protein functions are acetylation and phosphorylation.
An important role of acetylation is the regulation of DNA/RNA-protein interactions. A prominent example for this are histones, whose tail regions are lysine-rich and can be highly acetylated at their N-terminal domain. In spite of the utmost importance of this PTM, methods that allow the accurate measuring the site-specific acetylation degree are missing. One of the challenges in quantifying the acetylation degree at an individual lysine residue of the histones N-termini is the occurrence of multiple lysines in close proximity. Herein, we describe the development of the ”Fragment Ion Patchwork Quantification,” a new mass spectrometry-based approach for the highly accurate quantification of sites-pecific acetylation degrees. This method combines 13C1-acetyl derivatization on the protein level, proteolysis by low-specificity proteases and quantification on the fragment ion level. Acetylation degrees are determined from the isotope patterns of acetylated b and y ions. We have shown that this approach allows determining the site-specific acetylation degrees of all lysine residues for all core histones of Trypanosoma brucei. In addition, we demonstrate the use of this approach to identify the substrate sites of histone acetyltransferases and to monitor the changes in acetylation of the histones of canonical nucleosome and transcription start site nucleosomes.
Phosphorylation is one of the most common and most important PTMs. The analysis of the human genome showed that there are about 518 kinases and more than 500,000 phosphorylation sites are believed to exist in the cellular proteome. Protein phosphorylation plays a crucial role in signaling many different cell processes, such as intercellular communication, cell growth, differentiation of proliferation and apoptosis. Whereas MS-based identification and relative quantification of singly phosphorylated peptides have been greatly improved during the last decade, and large-scale analysis of thousands of phosphopeptides can now be performed on a routine-base, the analysis of multi-phosphorylated peptides is still lagging vastly behind. The low pKa value of phosphate group and the associated negative charge are considered the major source of the problems with the analysis of
multi-phosphorylated peptides. These problems include the formation of phosphopeptide-metal complexes during liquid chromatography (e.g. Fe 3+), which leads to a drastic deterioration of the chromatographic properties of these peptides (peak tailing), the decreased ionization efficiencies of phosphorylated peptides compared to their unphosphorylated counterparts, the labile nature of phosphate during CID/HCD fragmentation, and the unsuitability of low-charged phosphopeptides for ETD fragmentation are the most important factors that hinder phosphorylation analysis by LC-MS/MS. Here we aimed to develop a method for improving the identification of multi-phosphorylated peptides as well as the localization of phosphorylation sites by charge-reversal derivatization of the phosphate groups. This method employs a carbodiimide-mediated phosphoramidation to converted the phosphates to stable aromatic phosphoramidates. This chemical modification of phosphosite(s) reversed the negative charge of the phosphate group(s) and increased the number of the positive charges within the phosphopeptide. This modification prevented the formation of phosphopeptide-metal ion complexes that dramatically decreases or completely diminishes the signal intensity of protonated phosphopeptides, specifically multi-phosphorylated peptides. Furthermore, the increased net charge the (phospho-)peptides made them suitable for ETD fragmentation, which generated a high number of fragment ions with high intensities that led to a better phosphopeptide identification and localization of phosphosite(s) with high confidence.
Interleukin-6 (IL-6), oncostatin M (OSM), leukaemia inhibitory factor (LIF) and cardiotrophin-1 (CT-1) are members of the IL-6-type cytokine family that is characterised by sharing the common receptor subunit gp130. While the involvement of these polypeptides in cell differentiation, cell survival, proliferation, apoptosis, inflammation, haematopoiesis, immune response and acute phase reaction has already been demonstrated, the description of their role in development and progression of cardiac hypertrophy is still rather limited. A model has been postulated that declares the transient expression of IL-6-type cytokines as protective, while a continuous cardiac secretion of these proteins seems to be rather harmful for the heart. Within the first part of the study (results 4.1, 4.2 and 4.3) it was shown that OSM induces hypertrophy of primary neonatal rat cardiomyocytes (NRCM), just as its related cytokines LIF, CT-1 and hIL-6/hsIL-6R (hsIL-6R, human soluble IL-6 receptor). Regarding the hypertrophic potentials the LIFR/gp130 utilising cytokines (hLIF, hOSM and hCT-1) are stronger inducers than the OSMR/gp130 utilising mOSM. Human IL-6/hsIL-6R which signals via a gp130 homodimer has the weakest hypertrophic effect. The thorough analysis of typical signalling pathways initiated by IL-6-type cytokines revealed that STAT3 phosphorylation at Y705 seems to be the most important hypertrophy promoting pathway. In addition and in contrast to published work, we clearly demonstrate that classical IL-6 signalling (upon pure IL-6 treatment) has no hypertrophic effect on cardiomyocytes, because they lack sufficient amounts of the membrane-bound IL-6R. This is also true for neonatal rat cardiac fibroblasts (NRCFB). Since these cells can also influence cardiac hypertrophy, signalling pathways and target genes were additionally examined in NRCFB in response to OSM, LIF and IL-6/sIL-6R. One of the key findings of this thesis is the selective change in expression of cytokines and receptors of the IL-6 family in both cell types upon IL-6-type cytokine stimulation. A striking difference between NRCM and NRCFB is the fact that the target gene induction in NRCM is of similar duration upon mOSM and hIL-6/hsIL-6R treatment, while hIL-6/hsIL-6R is capable of promoting the induction of OSMR and IL-6 significantly longer in NRCFB. By searching for transcription factors or intermediate cytokines which could be responsible for this difference, a strong correlation between increased Il6 transcription and amount of mRNA levels for C/EBPβ and C/EBPδ was observed in response to IL-6/sIL-6R stimulation. Interestingly, mOSM also mediates the induction of C/EBPβ and δ, but the initiation is significantly less efficient than in response to IL-6/sIL-6R. Therefore, we assume that mOSM stimulation fails to reach threshold values required for a prolonged IL-6 secretion. Since we additionally observe a slight IL-6R mRNA upregulation in NRCFB, we assume that the combination of IL-6, LIF, C/EBPβ, C/EBPδ and IL-6R expression might be responsible for the observed different kinetics with which IL-6 and OSM stimulate NRCFB. In addition to the aforementioned proteins, members of the renin-angiotensin system seem to support the IL-6-type cytokine mediated hypertrophy. Since it has already been shown that angiotensin II vice versa induces IL-6 expression in NRCM and NRCFB, this enhanced expression of AT1α and ACE could be of crucial interest for the hypertrophy supporting phenotype. The second part of the presented work dealt with the characterisation of the receptor complexes of rat OSM. The central question of this analysis was, whether rOSM, just like mOSM, only binds the type II (OSMR/gp130) receptor complex or is able to utilise the type II and type I (LIFR/gp130) receptor complex. Using different experimental approaches (knock-down of the OSMR expression by RNA interference, blocking of the LIFR by LIF-05, an antagonistic LIF variant, and generation of stably transfected Ba/F3 cells expressing the newly cloned rat OSMR/gp130 or LIFR/gp130 receptor complex) we can clearly show that rat OSM surprisingly utilises both, the type I and type II receptor complex. Therefore it closely mimics the human situation. Furthermore, rOSM displays cross-species activities and stimulates cells of human as well as murine origin. Its signaling capacities closely mimic those of human OSM in cell types of different origin in the way that strong activation of the JAK/STAT, the MAP kinase as well as the PI3K/Akt pathways can be observed. Therefore, the results obtained in the last section of this thesis clearly suggest that rat disease models would allow evaluation of the relevance of OSM for human biology much better than murine models.
Background
Several intracellular acting bacterial protein toxins of the AB-type, which are known to enter cells by endocytosis, are shown to produce channels. This holds true for protective antigen (PA), the binding component of the tripartite anthrax-toxin of Bacillus anthracis. Evidence has been presented that translocation of the enzymatic components of anthrax-toxin across the endosomal membrane of target cells and channel formation by the heptameric/octameric \(PA_{63}\) binding/translocation component are related phenomena. Chloroquine and some 4-aminoquinolones, known as potent drugs against Plasmodium falciparium infection of humans, block efficiently the \(PA_{63}\)-channel in a dose dependent way.
Methodology/Principal Findings
Here we demonstrate that related positively charged heterocyclic azolopyridinium salts block the \(PA_{63}\)-channel in the µM range, when both, inhibitor and \(PA_{63}\) are added to the same side of the membrane, the cis-side, which corresponds to the lumen of acidified endosomal vesicles of target cells. Noise-analysis allowed the study of the kinetics of the plug formation by the heterocycles. In vivo experiments using J774A.1 macrophages demonstrated that the inhibitors of \(PA_{63}\)-channel function also efficiently block intoxication of the cells by the combination lethal factor and \(PA_{63}\) in the same concentration range as they block the channels in vitro.
Conclusions/Significance
These results strongly argue in favor of a transport of lethal factor through the \(PA_{63}\)-channel and suggest that the heterocycles used in this study could represent attractive candidates for development of novel therapeutic strategies against anthrax.
Dendritic cells (DCs) can be sub-divided into various subsets that play specialized roles in priming of adaptive immune responses. Atherosclerosis is regarded as a chronic inflammatory disease of the vessel wall and DCs can be found in non-inflamed and diseased arteries. We here performed a systematic analyses of DCs subsets during atherogenesis. Our data indicate that distinct DC subsets can be localized in the vessel wall. In C57BL/6 and low density lipoprotein receptor-deficient (Ldlr−/−) mice, CD11c+ MHCII+ DCs could be discriminated into CD103− CD11b+F4/80+, CD11b+F4/80− and CD11b−F4/80− DCs and CD103+ CD11b−F4/80− DCs. Except for CD103− CD11b− F4/80− DCs, these subsets expanded in high fat diet-fed Ldlr−/− mice. Signal-regulatory protein (Sirp)-α was detected on aortic macrophages, CD11b+ DCs, and partially on CD103− CD11b− F4/80− but not on CD103+ DCs. Notably, in FMS-like tyrosine kinase 3-ligand-deficient (Flt3l−/−) mice, a specific loss of CD103+ DCs but also CD103− CD11b+ F4/80− DCs was evidenced. Aortic CD103+ and CD11b+ F4/80− CD103− DCs may thus belong to conventional rather than monocyte-derived DCs, given their dependence on Flt3L-signalling. CD64, postulated to distinguish macrophages from DCs, could not be detected on DC subsets under physiological conditions, but appeared in a fraction of CD103− CD11b+ F4/80− and CD11b+ F4/80+ cells in atherosclerotic Ldlr−/− mice. The emergence of CD64 expression in atherosclerosis may indicate that CD11b+ F4/80− DCs similar to CD11b+ F4/80+ DCs are at least in part derived from immigrated monocytes during atherosclerotic lesion formation. Our data advance our knowledge about the presence of distinct DC subsets and their accumulation characteristics in atherosclerosis, and may help to assist in future studies aiming at specific DC-based therapeutic strategies for the treatment of chronic vascular inflammation.
Mg\(^{2+}\) plays a vital role in platelet function, but despite implications for life-threatening conditions such as stroke or myocardial infarction, the mechanisms controlling [Mg\(^{2+}\)]i in megakaryocytes (MKs) and platelets are largely unknown. Transient receptor potential melastatin-like 7 channel (TRPM7) is a ubiquitous, constitutively active cation channel with a cytosolic α-kinase domain that is critical for embryonic development and cell survival. Here we report that impaired channel function of TRPM7 in MKs causes macrothrombocytopenia in mice (Trpm7\(^{fl/fl-Pf4Cre}\)) and likely in several members of a human pedigree that, in addition, suffer from atrial fibrillation. The defect in platelet biogenesis is mainly caused by cytoskeletal alterations resulting in impaired proplatelet formation by Trpm7\(^{fl/fl-Pf4Cre}\) MKs, which is rescued by Mg\(^{2+}\) supplementation or chemical inhibition of non-muscle myosin IIA heavy chain activity. Collectively, our findings reveal that TRPM7 dysfunction may cause macrothrombocytopenia in humans and mice.
Zinc (Zn2+) can modulate platelet and coagulation activation pathways, including fibrin formation. Here, we studied the (patho)physiological consequences of abnormal platelet Zn2+ storage and release. To visualize Zn2+ storage in human and mouse platelets, the Zn2+ specific fluorescent dye FluoZin3 was used. In resting platelets, the dye transiently accumulated into distinct cytosolic puncta, which were lost upon platelet activation. Platelets isolated from Unc13d−/− mice, characterized by combined defects of α/δ granular release, showed a markedly impaired Zn2+ release upon activation. Platelets from Nbeal2−/− mice mimicking Gray platelet syndrome (GPS), characterized by primarily loss of the α-granule content, had strongly reduced Zn2+ levels, which was also confirmed in primary megakaryocytes. In human platelets isolated from patients with GPS, Hermansky-Pudlak Syndrome (HPS) and Storage Pool Disease (SPD) altered Zn2+ homeostasis was detected. In turbidity and flow based assays, platelet-dependent fibrin formation was impaired in both Nbeal2−/− and Unc13d−/− mice, and the impairment could be partially restored by extracellular Zn2+. Altogether, we conclude that the release of ionic Zn2+ store from secretory granules upon platelet activation contributes to the procoagulant role of Zn2+ in platelet-dependent fibrin formation.
Megakaryocytes (MKs) release platelets into the lumen of bone marrow (BM) sinusoids while remaining to reside within the BM. The morphogenetic events of this complex process are still not fully understood. We combined confocal laser scanning microscopy with transmission and serial block-face scanning electron microscopy followed by 3D-reconstruction on mouse BM tissue sections. These analyses revealed that MKs in close vicinity to BM sinusoid (BMS) wall first induce the lateral retraction of CXCL12-abundant reticular (CAR) cells (CAR), followed by basal lamina (BL) degradation enabling direct MK-sinusoidal endothelial cells (SECs) interaction. Subsequently, an endothelial engulfment starts that contains a large MK protrusion. Then, MK protrusions penetrate the SEC, transmigrate into the BMS lumen and form proplatelets that are in direct contact to the SEC surface. Furthermore, such processes are induced on several sites, as observed by 3D reconstructions. Our data demonstrate that MKs in interaction with CAR-cells actively induce BMS wall alterations, including CAR-cell retraction, BL degradation, and SEC engulfment containing a large MK protrusion. This results in SEC penetration enabling the migration of MK protrusion into the BMS lumen where proplatelets that are adherent to the luminal SEC surface are formed and contribute to platelet release into the blood circulation.
Type 1 diabetes is an autoimmune disease that leads to the destruction of insulin-producing pancreatic beta cells and consequently to hyperglycemia. In the last 60 years, the prevalence of type 1 diabetes has been increasing constantly and is predicted to continue rising. About 80% of the disease risk is attributable to the genetic variation. Thanks to genome wide association studies the number of known disease-associated polymorphisms climbed from five to 53 in the last 10 years. As these studies reveal possible candidate genes but not underlying mechanisms we strove to take the next step and explore the association of two genes suggested by these studies with type 1 diabetes. As a method of choice we decided to use lentiviral RNAi in non obese diabetic (NOD) mice, a widely-used model for type 1 diabetes, introducing a shRNA directed against the target message into the genome of this mouse strain via a lentivirus. This allowed us to study the partial loss-of-function of the target gene within the context of diabetes, directly seeing its effect on autoimmune mechanisms. In this thesis we examined two different genes in this manner, Ctla4 and Clec16a. A type 1 diabetes associated polymorphism in the CTLA4 gene had been found to alter the splicing ratio of its variants soluble CTLA-4 (sCTLA-4) and full length CTLA-4, the associated allele producing less sCTLA-4 than the protective allele. We mimicked this effect by specifically targeting the sCtla4 mRNA via lentiviral RNAi in the NOD model. As a result we could confirm the reduction of sCTLA-4 to accelerate type 1 diabetes development. Furthermore we could show a function of sCTLA-4 in regulatory T cells, more specifically at least partly in their ability to modulate costimulation by antigen presenting cells. The second candidate gene, Clec16a was targeted with the shRNA in a way that was designed to knock down most splice variants. As the gene function and the effect of the associated SUMMARY 10 polymorphism was unknown, we reasoned this method to be feasible to investigate its role in type 1 diabetes. The knockdown of Clec16a in NOD mice resulted in an almost complete protection from diabetes development that could be attributed to T cells dysfunction. However, as expression patterns and a study of the Drospophila orthologue suggested a possible role of CLEC16A in antigen presentation we also examined antigen presenting cells in the thymus and periphery. Although we did not detect any effect of the knockdown on peripheral antigen presenting cells, thymic epithelial cells were clearly affected by the loss of CLEC16A, rendering them more activated and shifting the ratio of cortical to medullary epithelial cells in favor of cortical cells. We therefore suggest a role of CLEC16A in the selection of T cells, that needs, however, to be further investigated. In this thesis we provided a feasible and fast method to study function of genes and even of single splice variants within the NOD mouse model. We demonstrate its usefulness on two candidate genes associated with type 1 diabetes by confirming and unraveling the cause of their connection to the disease.