title: The Size of Activating and Inhibitory Killer Ig-like Receptor Nanoclusters Is Controlled by the Transmembrane Sequence and Affects Signaling.
abstract: Super-resolution microscopy has revealed that immune cell receptors are organized in nanoscale clusters at cell surfaces and immune synapses. However, mechanisms and functions for this nanoscale organization remain unclear. Here, we used super-resolution microscopy to compare the surface organization of paired killer Ig-like receptors (KIR), KIR2DL1 and KIR2DS1, on human primary natural killer cells and cell lines. Activating KIR2DS1 assembled in clusters two-fold larger than its inhibitory counterpart KIR2DL1. Site-directed mutagenesis established that the size of nanoclusters is controlled by transmembrane amino acid 233, a lysine in KIR2DS1. Super-resolution microscopy also revealed two ways in which the nanoscale clustering of KIR affects signaling. First, KIR2DS1 and DAP12 nanoclusters are juxtaposed in the resting cell state but coalesce upon receptor ligation. Second, quantitative super-resolution microscopy revealed that phosphorylation of the kinase ZAP-70 or phosphatase SHP-1 is favored in larger KIR nanoclusters. Thus, the size of KIR nanoclusters depends on the transmembrane sequence and affects downstream signaling.
authors.affiliation: Manchester Collaborative Centre for Inflammation Research, University of Manchester, 46 Grafton Street, Manchester M13 9NT, UK.
authors.forename: Anna
authors.initials: A
authors.lastname: Oszmiana
authors.affiliation: Manchester Collaborative Centre for Inflammation Research, University of Manchester, 46 Grafton Street, Manchester M13 9NT, UK.
authors.forename: David J
authors.initials: DJ
authors.lastname: Williamson
authors.affiliation: Division of Cell and Molecular Biology, Sir Alexander Fleming Building, Imperial College London, London SW7 2AZ, UK.
authors.forename: Shaun-Paul
authors.initials: SP
authors.lastname: Cordoba
authors.affiliation: Manchester Collaborative Centre for Inflammation Research, University of Manchester, 46 Grafton Street, Manchester M13 9NT, UK.
authors.forename: David J
authors.initials: DJ
authors.lastname: Morgan
authors.affiliation: Manchester Collaborative Centre for Inflammation Research, University of Manchester, 46 Grafton Street, Manchester M13 9NT, UK.
authors.forename: Philippa R
authors.initials: PR
authors.lastname: Kennedy
authors.affiliation: Manchester Collaborative Centre for Inflammation Research, University of Manchester, 46 Grafton Street, Manchester M13 9NT, UK.
authors.forename: Kevin
authors.initials: K
authors.lastname: Stacey
authors.affiliation: Manchester Collaborative Centre for Inflammation Research, University of Manchester, 46 Grafton Street, Manchester M13 9NT, UK. Electronic address: daniel.davis@manchester.ac.uk.
authors.forename: Daniel M
authors.initials: DM
authors.lastname: Davis
mesh_terms: D048868:Adaptor Proteins, Signal Transducing / Q000378:metabolism; D019943:Amino Acid Substitution; D002460:Cell Line; D002462:Cell Membrane / Q000378:metabolism*; D002999:Clone Cells; D006801:Humans; D007694:Killer Cells, Natural / Q000378:metabolism; D008565:Membrane Proteins / Q000378:metabolism; D053758:Nanoparticles / Q000737:chemistry*; D010766:Phosphorylation; D011485:Protein Binding; D053824:Protein Tyrosine Phosphatase, Non-Receptor Type 6 / Q000378:metabolism; D054340:Receptors, KIR / Q000378:metabolism*; D054342:Receptors, KIR2DL1 / Q000378:metabolism*; D015398:Signal Transduction; D051746:ZAP-70 Protein-Tyrosine Kinase / Q000378:metabolism
chemical_list: D048868:Adaptor Proteins, Signal Transducing; C093100:KIR2DL1 protein, human; C496212:KIR2DS1 protein, human; D008565:Membrane Proteins; D054340:Receptors, KIR; D054342:Receptors, KIR2DL1; C110546:TYROBP protein, human; D051746:ZAP-70 Protein-Tyrosine Kinase; C497261:ZAP70 protein, human; D053824:Protein Tyrosine Phosphatase, Non-Receptor Type 6