Abstract
The organization and dynamics of receptors and other molecules in the plasma membrane are not well understood. Here we analyzed the spatio-temporal dynamics of T cell antigen receptor (TCR) complexes and linker for activation of T cells (Lat), a key adaptor molecule in the TCR signaling pathway, in T cell membranes using high-speed photoactivated localization microscopy, dual-color fluorescence cross-correlation spectroscopy and transmission electron microscopy. In quiescent T cells, both molecules existed in separate membrane domains (protein islands), and these domains concatenated after T cell activation. These concatemers were identical to signaling microclusters, a prominent hallmark of T cell activation. This separation versus physical juxtapositioning of receptor domains and domains containing downstream signaling molecules in quiescent versus activated T cells may be a general feature of plasma membrane-associated signal transduction.
Publication types
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Research Support, N.I.H., Extramural
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Research Support, Non-U.S. Gov't
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Research Support, U.S. Gov't, Non-P.H.S.
MeSH terms
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Adaptor Proteins, Signal Transducing / genetics
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Adaptor Proteins, Signal Transducing / metabolism*
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Animals
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Cell Membrane / metabolism*
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Cell Membrane / ultrastructure
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Genetic Vectors / genetics
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Green Fluorescent Proteins / genetics
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Green Fluorescent Proteins / metabolism
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Kinetics
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Lymphocyte Activation / immunology
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Membrane Microdomains / metabolism
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Membrane Microdomains / ultrastructure
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Membrane Proteins / genetics
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Membrane Proteins / metabolism*
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Mice
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Microscopy, Electron, Transmission
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Microscopy, Fluorescence / methods
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Models, Biological
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Phosphoproteins / genetics
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Phosphoproteins / metabolism*
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Protein Transport
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Receptors, Antigen, T-Cell / genetics
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Receptors, Antigen, T-Cell / metabolism*
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Recombinant Fusion Proteins / genetics
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Recombinant Fusion Proteins / metabolism
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Retroviridae / genetics
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T-Lymphocytes / cytology
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T-Lymphocytes / immunology
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T-Lymphocytes / metabolism*
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T-Lymphocytes / ultrastructure
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Transfection
Substances
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Adaptor Proteins, Signal Transducing
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Lat protein, mouse
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Membrane Proteins
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Phosphoproteins
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Receptors, Antigen, T-Cell
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Recombinant Fusion Proteins
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Green Fluorescent Proteins