A5042729336- Cellular transport and secretion
- Immune Response and Inflammation
- Lipid Membrane Structure and Behavior
- Immune Cell Function and Interaction
- CAR-T cell therapy research
- T-cell and B-cell Immunology
- interferon and immune responses
- Advanced Fluorescence Microscopy Techniques
- Monoclonal and Polyclonal Antibodies Research
- Immune cells in cancer
- Cell Image Analysis Techniques
- Supramolecular Self-Assembly in Materials
- RNA Interference and Gene Delivery
- Receptor Mechanisms and Signaling
- Cancer Immunotherapy and Biomarkers
- Retinal Development and Disorders
- Neutrophil, Myeloperoxidase and Oxidative Mechanisms
- Advanced biosensing and bioanalysis techniques
- Photoreceptor and optogenetics research
- Medical Imaging and Analysis
- Fungal and yeast genetics research
- Posttraumatic Stress Disorder Research
- Immunotherapy and Immune Responses
- Autophagy in Disease and Therapy
- bioluminescence and chemiluminescence research
Max Planck Institute for Infection Biology
2019-2025
Manchester University NHS Foundation Trust
2025
University of Manchester
2025
Charité - Universitätsmedizin Berlin
2020-2023
University of California, San Francisco
2013-2017
Howard Hughes Medical Institute
2017
National Centre for Biological Sciences
2016-2017
Woodwell Climate Research Center
2016-2017
MRC Laboratory of Molecular Biology
2008-2012
Medical Research Council
2011-2012
Programmed cell death-1 (PD-1) is a coinhibitory receptor that suppresses T activation and an important cancer immunotherapy target. Upon by its ligand PD-L1, PD-1 thought to suppress signaling through the (TCR). By titrating in biochemical reconstitution system, we demonstrate co-receptor CD28 strongly preferred over TCR as target for dephosphorylation PD-1-recruited Shp2 phosphatase. We also show CD28, but not TCR, preferentially dephosphorylated response PD-L1 intact system. These results...
Dual colour total internal reflection fluorescence microscopy is a powerful tool for decoding the molecular dynamics of clathrin-mediated endocytosis (CME). Typically, recruitment fluorescent protein-tagged endocytic protein was referenced to disappearance spot-like clathrin-coated structure (CCS), but precision CCS as marker canonical CME remained unknown. Here we have used an imaging assay based on detect scission events with resolution ∼ 2 s. We found that engulfed comparable amounts...
Myosin VI plays a role in the maintenance of Golgi morphology and exocytosis. In yeast 2-hybrid screen we identified optineurin as binding partner for myosin at complex confirmed this interaction range protein studies. Both proteins colocalize vesicles plasma membrane. When is depleted from cells using RNA interference, lost complex, fragmented exocytosis vesicular stomatitis virus G-protein to membrane dramatically reduced. Two further partners have been identified: huntingtin Rab8. We show...
Clathrin-mediated endocytosis proceeds by a sequential series of reactions catalyzed discrete sets protein machinery. The final reaction in clathrin-mediated is membrane scission, which mediated the large guanosine triophosphate hydrolase (GTPase) dynamin and may involve actin-dependent recruitment N-terminal containing BIN/Amphiphysin/RVS domain (N-BAR) proteins. Optical microscopy has revealed detailed picture when where particular types are recruited ∼20–30 s preceding scission....
Current knowledge of the structural changes taking place during clathrin-mediated endocytosis is largely based on electron microscopy images fixed preparations and x-ray crystallography data purified proteins. In this paper, we describe a study clathrin-coated pit dynamics in living cells using ion conductance to directly image shape, combined with simultaneous confocal follow molecule-specific fluorescence. We find that 70% pits closed formation protrusion grew one side pit, covered entire...
Abstract We developed a system for optogenetic release of single molecules in cells. confined soluble and transmembrane proteins to the Golgi apparatus via photocleavable protein released them by short pulses light. Our method allows light dose-dependent delivery functional cytosol plasma membrane amounts compatible with single-molecule imaging, greatly simplifying access microscopy any live were able reconstitute ion conductance delivering BK LRRC8/volume-regulated anion channels membrane....
EpsinR is a clathrin-coated vesicle (CCV)-associated protein that binds to vti1b, suggesting it may be vti1b-selective adaptor. Depletion of epsinR undetectable levels in HeLa cells using siRNA causes vti1b redistribute from the perinuclear region cell periphery, but vti1a also redistributes epsinR-depleted cells, and both vti isoforms AP-1-depleted cells. As more direct assay for function, we isolated CCVs control siRNA-treated then looked differences cargo content. In clathrin-depleted...
The AP-2 adaptor complex plays a key role in cargo recognition and clathrin-coated vesicle formation at the plasma membrane. To investigate functions of individual binding sites domains vivo, we have stably transfected HeLa cells with wild-type mutant small interfering RNA-resistant alpha mu2 subunits then used siRNA knockdowns to deplete endogenous proteins. Mutating PtdIns(4,5)P2 site alpha, phosphorylation mu2, or YXXPhi impairs function, as assayed by transferrin uptake. In contrast,...
A recurring feature of innate immune receptor signaling is the self-assembly proteins into oligomeric complexes. The Myddosome an complex that required to transmit inflammatory signals from TLR/IL1Rs and consists MyD88 IRAK family kinases. However, molecular basis for how self-assemble regulate intracellular remains poorly understood. Here, we developed a novel assay analyze spatiotemporal dynamics IL1R in live cells. We found oligomerization inducible initially reversible. Moreover,...
Abstract Programmed death-1 (PD-1) is a co-inhibitory receptor that suppresses T cell activation and an important cancer immunotherapy target. Upon by its ligand PD-L1, PD-1 thought to suppress signaling through the (TCR). Here, titrating strength of in both biochemical reconstitution systems cells, we demonstrate coreceptor CD28 strongly preferred over TCR as target for dephosphorylation PD-1- recruited Shp2 phosphatase. We also show colocalizes with costimulatory plasma membrane...
Anatomical RS9 segmentectomy is challenging due the intricate intersegmental planes (ISPs) and varied anatomical structures involved, with few studies having robustly assessed three-dimensional (3D) reconstruction techniques for anatomy surgical procedures of segment. This study aimed to analyze application 3D vascular bronchial branching patterns right anterior, lateral, posterior basal segments (RS8, RS9, RS10, respectively) segmentectomy. From May 2020 2022, reconstructions 354 patients...
Protein polymer scaffolds composed of death fold (DF) proteins are critical to the formation signalosomes in immune signaling. The biophysical properties that these polymeric require for signal transduction not clearly defined. Here, we engineered single-component DF signalosomes. We found functionality depends on stability provided by polymer, which could also be achieved with a bacterial domain, synthetic filament-forming and amyloid-like sequences. This demonstrates importance...
IL-1 receptor (IL-1R) signaling can activate thresholded invariant outputs and proportional that scale with the amount of stimulation. Both responses require Myddosome, a multiprotein complex. The Myddosome is required for polyubiquitin chain formation NF-kB signaling. However, how these signals are spatially temporally regulated to drive switch-like outcomes not understood. During IL-1R signaling, Myddosomes dynamically reorganize into multi-Myddosome clusters at cell membrane. Blockade...
The AP‐1 and AP‐2 complexes are the most abundant adaptors in clathrin‐coated vesicles (CCVs), but clathrin‐mediated trafficking can still occur absence of any detectable or AP‐2. To find out whether adaptor abundance reflects cargo abundance, we used lectin pulldowns to identify major membrane glycoproteins CCVs from human placenta rat liver. Both preparations contained three prominent high molecular‐weight proteins: cation‐independent mannose 6‐phosphate receptor (CIMPR), carboxypeptidase...
Abstract A key feature of innate immune signaling is the compartmentalization effectors into cellular structures referred to as signalosomes. Critical formation these compartments are protein polymers composed Death Domains (DD). However, biophysical properties polymeric scaffolds require for signal transduction not clearly defined. Here, we engineered a single-component signalosome, Chimeric Higher-order Assemblies Receptor Mediated Signaling (CHARMS). We found that CHARMS functionality...
Abstract T cells mount an immune response by measuring the binding strength of its cell receptor (TCR) for peptide-loaded MHCs (pMHC) on antigen-presenting cell. How convert lifetime extracellular TCR-pMHC interaction into intracellular signal remains unknown. Here, we developed a synthetic signaling system in which domains TCR and pMHC were replaced with short hybridizing strands DNA. Remarkably, can discriminate between DNA ligands differing single base pair. Single molecule imaging...
Extended abstract of a paper presented at Microscopy and Microanalysis 2011 in Nashville, Tennessee, USA, August 7–August 11, 2011.
Abstract Signaling pathways can produce digital invariant outputs and analog that scale with the amount of stimulation. In IL-1 receptor (IL-1R) signaling both types require Myddosome, a multi-protein complex. The Myddosome is required for polyubiquitin chain formation NF-kB signaling. However, ways in which these signals are spatially temporally regulated to drive switch-like proportional outcomes not understood. We find during IL-1R signaling, Myddosomes dynamically re-organize into large,...
Abstract The controlled oligomerization of signaling proteins is an essential feature many inflammatory pathways. An example IL-1 receptor signaling, which relies on the Death Domain (DD)-containing MyD88 and IRAK family kinases. This process leads to assembly Myddosome complex, disrupting holds potential for anti-inflammatory treatments. However, IRAKs’ activity also regulated by auto-/trans-phosphorylation, it unclear if these processes operate at or downstream assembly. Here, we find that...
Abstract A recurring feature of innate immune receptor signaling is the self-assembly proteins into oligomeric complexes. The Myddosome an complex that required to transmit inflammatory signals from TLR/IL1Rs and consists MyD88 IRAK family kinases. However, molecular basis for how self-assemble regulate intracellular remains poorly understood. Here, we developed a novel assay analyze spatiotemporal dynamics IL1R in live cells. We found oligomerization inducible initially reversible....
Abstract A recurring feature of innate immune receptor signaling is the self-assembly proteins into oligomeric complexes. The Myddosome an complex that required to transmit inflammatory signals from TLR/IL1Rs and consists MyD88 IRAK family kinases. However, molecular basis for how self-assemble regulate intracellular remains poorly understood. Here, we developed a novel assay analyze spatiotemporal dynamics IL1R in live cells. We found oligomerization inducible initially reversible....