• We found that GML potently altered order and disorder dynamics in the plasma membrane that resulted in reduced formation of LAT, PLC-γ, and AKT microclusters. (nature.com)
  • Considerable controversy arose over the concept that cholesterol/sphingolipid-rich rafts in the T cell plasma membrane serve as a platform for TCR signalling reactions. (biomedcentral.com)
  • Here we review new biophysical and biochemical data which provide an updated picture of the highly dynamic nanometer-sized cholesterol/sphingolipid-rich raft domains stabilised by protein-networks to form TCR signalling platforms in the T cell plasma membrane. (biomedcentral.com)
  • Early T cell signalling reactions are embedded in the complex and dynamic lipid bilayer matrix of the T cell plasma membrane and are critically defined by their lateral compartmentalisation in plasma membrane domains [ 1 , 2 ]. (biomedcentral.com)
  • We will here discuss recent data which provide a detailed picture of the membrane raft characteristics of the plasma membrane domains supporting active TCR signalling protein complexes. (biomedcentral.com)
  • The TCR signalling cascade is initiated by phosphorylation of critical tyrosines of the TCR/CD3 complex by the Src-family kinases Lck or Fyn which are anchored in the cytoplasmic leaflet of the T cell plasma membrane. (biomedcentral.com)
  • The cytosolic ZAP-70 tyrosine kinase is recruited to the T cell plasma membrane via binding to the tyrosine phosphorylated TCR-complex [ 3 ]. (biomedcentral.com)
  • These complexes assemble into submicron TCR signalling domains in the T cell plasma membrane which were first studied by confocal fluorescence microscopy at the contact zone of a T cell with TCR-activating glass coverslips. (biomedcentral.com)
  • Video microscopy technology resolving single molecule movement was employed to monitor the dynamic interaction of membrane-associated signalling proteins with TCR signalling clusters in plasma membrane domains. (biomedcentral.com)
  • These studies tracked the movement of Lck and LAT in the T cell plasma membrane and showed retention of these proteins in the submicron TCR signalling domains and their subsequent release. (biomedcentral.com)
  • However specific lipid-mediated interactions at the T cell plasma membrane bilayer are also a central functional element in early TCR signalling. (biomedcentral.com)
  • Numerous intracellular signalling proteins interact with plasma membrane lipids of the cytoplasmic leaflet via specific lipid headgroup-binding domains. (biomedcentral.com)
  • Confocal fluorescence microscopy was employed to monitor the distribution of TCR signalling plasma membrane sites and to relate their distribution to that of clustered raft markers which show resistance to Triton X 100 detergent solubilisation. (biomedcentral.com)
  • We detail enzyme-linked immunosorbent assays to determine relative GPCR plasma membrane and total expression levels. (bvsalud.org)
  • Altered membrane events induced selective inhibition of TCR-induced phosphorylation of regulatory P85 subunit of PI3K and AKT as well as abrogated calcium influx. (nature.com)
  • Moreover, we identify a distinct, N-terminal, higher affinity interaction interface between LRRK2 phosphorylated Rab8 and Rab10 termed 'Site #2', that can retain LRRK2 on membranes in cells to catalyze multiple, subsequent phosphorylation events. (stanford.edu)
  • Proteins which are found in membranes including cellular and intracellular membranes. (lookformedical.com)
  • The intracellular transfer of information (biological activation/inhibition) through a signal pathway. (lookformedical.com)
  • Those coupled to membrane depolarization or intracellular release of calcium include the receptor-mediated activation of cytotoxic functions in granulocytes and the synaptic potentiation of protein kinase activation. (lookformedical.com)
  • This protocol describes steps for cloning HA- and FLAG-tagged GPCRs, transient transfection of CHO-K1 or HEK293-T cells, and determination of basal and ligand-induced changes in intracellular cAMP levels. (bvsalud.org)
  • These interactions and their essential functional consequences for cell surface receptor signalling reactions are excellently reviewed in [ 9 ]. (biomedcentral.com)
  • I am committed to interdisciplinary research, and an example of the successes gained from this approach is the project with Prof A Nabok (Engineering Sheffield Hallam University) using total internal reflection ellipsometry to quantify protein-membrane interactions on native plant membranes and human cell lines. (brookes.ac.uk)
  • Program access grant to the STFC Harwell Laser Facility 'The Plant Cell Initiative: Protein interactions in the higher plant secretory pathway' 2017-2021 (approximate value of £200K). (brookes.ac.uk)
  • Examples of signal transduction systems are the GAMMA-AMINOBUTYRIC ACID-postsynaptic receptor-calcium ion channel system, the receptor-mediated T-cell activation pathway, and the receptor-mediated activation of phospholipases. (lookformedical.com)
  • A feed-forward pathway drives LRRK2 kinase membrane recruitment and activation. (stanford.edu)
  • To address these knowledge gaps, we treated human activated peripheral blood T cells (APBTs) with solubilized GML or ethanol vehicle control. (nature.com)
  • They are a highly dynamic continuum of vesicles with a rich variety of established biochemical roles in cells, to which Ca2+ storage can now be added. (wikipedia.org)
  • A molecular understanding of membrane traffic has broad implications for our understanding of growth control in cancer, receptor trafficking errors in heart disease, regulation of insulin secretion in diabetes and synaptic vesicle biogenesis and transport in neurological disorders. (stanford.edu)
  • The analysis of detergent-resistant T cell membranes had initiated the concept that early TCR signalling steps take place in cholesterol/sphingolipid-rich raft domains of T cell plasma membranes [ 10 ] (see Box for an overview of the current perception of raft domains in cell membranes). (biomedcentral.com)
  • GML treated T cells have decreased cellular proliferation when stimulated with phorbol 12-myristate 12-acetate (PMA), concanavalin A (ConA), and Toxic Shock Syndrome Toxin-1 (TSST-1) 13 . (nature.com)
  • Signal transduction plays an important role in activating cellular functions, cell differentiation, and cell proliferation. (lookformedical.com)
  • Subsequently, NAADP has been detected in sources as diverse as human sperm, red and white blood cells, liver, and pancreas, to name but a few. (wikipedia.org)
  • Subsequently, other cell types have followed suit, as exemplified by the pancreas (acinar and beta cells), T-cells, and smooth muscle. (wikipedia.org)
  • However, how GML treatment affects the full T cell activation response from signaling events to effector functions is not well understood. (nature.com)
  • The T cell antigen receptor (TCR) provides the key signal for activation of T lymphocytes to perform their numerous effector functions in adaptive immune responses. (biomedcentral.com)
  • The transduction mechanisms that couple cell stimuli to such NAADP increases are ill-defined, with some suggestions of cyclic AMP or cytosolic Ca2+ itself stimulating synthesis. (wikipedia.org)
  • Revisiting the Latency of Uridine Diphosphate-Glucuronosyltransferases (UGTs)-How Does the Endoplasmic Reticulum Membrane Influence Their Function? (mdpi.com)
  • In this study, we have mechanistically examined if GML affects the signaling and functional output of human primary T cells. (nature.com)
  • To directly address these questions, we mechanistically characterized how GML affects human primary T cell activation. (nature.com)
  • These studies suggest that GML potentially alters T cell activation. (nature.com)
  • In addition, the mechanism behind how GML mediates suppressed T cell activation is completely unknown. (nature.com)
  • However, no studies have comprehensively examined the effects of TCR-mediated T cell activation or identified potential mechanisms for the effects of GML on TCR-mediated T cell activation. (nature.com)
  • Temperature optimization studies were carried out using HEK293H cells transiently transfected with the adenosine receptor A2a and the GloSensor™ plasmid (pGloSensor-20F or -22F). (bvsalud.org)
  • Certainly NAADP production can occur in vitro but whether it occurs in vivo is another question (because genetic knockout or knock-down of ADP-ribosyl cyclases has no effect on NAADP production in some cell types), and there may be other routes which require different substrates and enzymes. (wikipedia.org)
  • APBTs were treated with varying doses of GML for 24 hours and live cells were counted using tryphan blue staining. (nature.com)
  • We also present a detailed protocol for monitoring cAMP levels in live cells expressing endogenous or exogenous GPCRs. (bvsalud.org)
  • Cells propagated in vitro in special media conducive to their growth. (lookformedical.com)
  • T cells become activated upon engagement of their TCRs by a cognate peptide-MHC ligand presented on the surface of an antigen-presenting or target cell. (biomedcentral.com)
  • Single pavement layer of cells which line the luminal surface of the entire vascular system and regulate the transport of macromolecules and blood components. (lookformedical.com)