A5035690049- Cellular transport and secretion
- Platelet Disorders and Treatments
- Lipid Membrane Structure and Behavior
- Complement system in diseases
- Cell Adhesion Molecules Research
- Blood Coagulation and Thrombosis Mechanisms
- Erythrocyte Function and Pathophysiology
- Calcium signaling and nucleotide metabolism
- Lysosomal Storage Disorders Research
- Blood groups and transfusion
- Antiplatelet Therapy and Cardiovascular Diseases
- Myeloproliferative Neoplasms: Diagnosis and Treatment
- RNA Interference and Gene Delivery
- Blood properties and coagulation
- Endoplasmic Reticulum Stress and Disease
- Monoclonal and Polyclonal Antibodies Research
- melanin and skin pigmentation
- RNA regulation and disease
- Photosynthetic Processes and Mechanisms
- Electron Spin Resonance Studies
- Microtubule and mitosis dynamics
- Advanced NMR Techniques and Applications
- Mast cells and histamine
- NMR spectroscopy and applications
- Venous Thromboembolism Diagnosis and Management
Florida Aquarium
2023
University College London
2013-2022
MRC Laboratory for Molecular Cell Biology
2013-2022
University of Glasgow
2019
Indian Institute of Technology Bombay
2019
Tohoku University
2019
Universidad de Zaragoza
2019
Bielefeld University
2019
University of Edinburgh
2019
Queen Mary University of London
1957-2018
Specialized cells exploit adaptor protein complexes for unique post-Golgi sorting events, providing a model system to specify function. Here, we show that AP-3 and AP-1 function independently in of the melanocyte-specific tyrosinase from endosomes melanosome, specialized lysosome-related organelle distinguishable lysosomes. localize melanocytes primarily clathrin-coated buds on tubular early near melanosomes. Both adaptors recognize dileucine-based melanosome signal, largely colocalizes with...
The study of actin in regulated exocytosis has a long history with many different results numerous systems. A major limitation on identifying precise mechanisms been the paucity experimental systems which function directly assessed alongside granule content release at distinct steps single secretory organelle sufficient spatiotemporal resolution. Using dual-color confocal microscopy and correlative electron human endothelial cells, we visually distinguished two sequential...
Weibel-Palade bodies (WPBs), endothelial-specific secretory granules that are central to primary hemostasis and inflammation, occur in dimensions ranging between 0.5 5 μm. How their size is determined whether it has a functional relevance at present unknown. Here, we provide evidence for dual role of the Golgi apparatus controlling these carriers. At ministack level, cisternae constrain nanostructures ("quanta") von Willebrand factor (vWF), main WPB cargo. The ribbon architecture then allows...
It is well established that a proportion of newly synthesized lysosomal enzymes and class II major histocompatibility complex antigens are delivered directly to the endocytic pathway from Golgi complex. Here we show significant transferrin receptors can be detected in endosomes before reaching cell surface. These endosome more efficiently than either constitutively secreted soluble proteins or glycophosphatidylinositol-anchored plasma membrane suggesting their transfer signal-dependent....
Ligands that bind mammalian cell surface integrins with high affinity can mediate cellular internalization. We show particles of the bacteriophage fd display cyclic integrin-binding peptide sequence GGCRGDMFGC in a proportion their major coat protein subunits to cells and are efficiently internalized. In displayed conformation RGD motif is restricted within hairpin loop formed by disulfide bridge between 2 cysteine residues. Cellular internalization phage was demonstrated confocal...
We have developed a novel technique with which to investigate the morphological basis of exocytotic traffic. used expression HRP from cDNA in variety cells combination peroxidase cytochemistry outline traffic into and out Golgi apparatus at electron microscopic level very high sensitivity. A secretory form (ssHRP) is active beginning pathway activity efficiently cleared cells. Investigation elements involved itinerary soluble ER proteins using ssHRP tagged retention motif (ssHRPKDEL) shows...
Clathrin provides an external scaffold to form small 50–100-nm transport vesicles. In contrast, formation of much larger dense-cored secretory granules is driven by selective aggregation internal cargo at the trans-Golgi network; only known role clathrin in remove missorted proteins small, coated vesicles during maturation these spherical organelles. The Weibel-Palade bodies (WPBs) also driven, but are cigar-shaped organelles up 5 μm long. We hypothesized that a cytoplasmic coat might be...
The transfer of newly synthesized membrane proteins moving from the rough endoplasmic reticulum (RER) to Golgi complex has been studied by electron microscopy in HEp-2 cells transfected with cDNAs for chimeric proteins. These consist a reporter enzyme, horseradish peroxidase (HRP), anchored transmembrane domains two integral proteins, transferrin receptor and sialyl-transferase. chimeras are distributed throughout nuclear envelope, RER, vesicular tubular clusters (VTCs) network tubules...
The identification of organelles is crucial for efficient cellular function, yet the basic underlying mechanisms by which this might occur have not been established. One group proteins likely to be central organelle identity Rab family small GTPases. We thus investigated recruitment membranes using endothelial cells as a model system. report that Weibel-Palade bodies, Von Willebrand Factor storage compartment human umbilical vein cells, contain Rab27a. also found body-like structures induced...
The Weibel-Palade bodies (WPBs) of endothelial cells play an important role in haemostasis and the initiation inflammation, yet their biogenesis is poorly understood. Tubulation major content protein, von Willebrand factor (VWF), crucial to WPB function, so we investigated further relationship between VWF tubule formation human umbilical vein (HUVECs). By using high-pressure freezing freeze substitution before electron microscopy, visualised tubules trans-Golgi network (TGN), as well...
The neuronal ceroid lipofuscinoses (NCLs) are common neurodegenerative disorders of childhood and classified as lysosomal storage diseases since affected cells exhibit lysosomes containing lipofuscin-like material. CLN3 is the most widely conserved NCL gene, suggesting that it has a basic eukaryotic cell function; its loss might be expected to cause earliest onset and/or severe disease. However, mutations in linked juvenile (JNCL), latest mildest form children. We sought explain this...
The neuronal ceroid lipofuscinoses (NCLs, Batten disease) are a group of inherited childhood-onset neurodegenerative disorders characterized by the lysosomal accumulation undigested material within cells. To understand this dysfunction, we analysed trafficking cation-independent mannose 6-phosphate receptor (CI-MPR), which delivers digestive enzymes to lysosomes. A common form NCL is caused mutations in CLN3, multipass transmembrane protein unknown function. We report that ablation CLN3...
Von-Willebrand factor (vWF) is a highly multimerized hemostatic glycoprotein that stored in endothelial Weibel-Palade bodies (WPB) and secreted upon cell stimulation to act recruiting platelets sites of vessel injury. Only fully matured multimeric vWF represents an efficient anchor for platelets, cells have developed mechanisms prevent release immature vWF. Full maturation occurs within WPB following their translocation from perinuclear site emergence at the trans-Golgi network (TGN)...