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.
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29 January 2010
In the version of this article initially published, some rows in Table 1 were misaligned. The error has been corrected in the HTML and PDF versions of the article.
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Acknowledgements
We thank B. Wilson for advice on TEM and plasma membrane sheet preparation; J. Perrino from the Cell Sciences Imaging Facility for expertise and service; and F. Tynan for comments on the manuscript. Supported by the US National Institutes of Health (AI 55277 to M.M.D.), the US National Science Foundation (J.T.G.), the Howard Hughes Medical Institute (M.M.D. and J.T.G.) and the Human Frontier Science Program (B.F.L.).
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B.F.L. and M.M.D., conceptualization and manuscript preparation; B.F.L., PALM experiments and analysis, TEM studies, molecular biology, cell biology and protein chemistry; M.A.M., PALM conceptualization and analysis software; M.B.F. and J.T.G., dcFCCS conceptualization; M.B.F., dcFCCS software and analysis; M.B.F. and B.F.L., dcFCCS experiments; and J.B.H., anti-TCRβ-scFv and microscope setup.
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Supplementary Figures 1–11, Supplementary Tables 1–2 and Supplementary Methods (PDF 8659 kb)
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Lillemeier, B., Mörtelmaier, M., Forstner, M. et al. TCR and Lat are expressed on separate protein islands on T cell membranes and concatenate during activation. Nat Immunol 11, 90–96 (2010). https://doi.org/10.1038/ni.1832
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DOI: https://doi.org/10.1038/ni.1832
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