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TCR and Lat are expressed on separate protein islands on T cell membranes and concatenate during activation

Nature Immunology volume 11, pages 90–96 (2010)Cite this article

An Erratum to this article was published on 01 June 2010

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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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Figure 1: Analysis of the distribution of CD3ζ and Lat in native T cell plasma membrane sheets by hsPALM.
Figure 2: Analysis of the distribution of endogenous TCRβ in T cells by hsPALM.
Figure 3: TEM analysis of the concatenation of CD3ζ and Lat islands.
Figure 4: Analysis of the correlation of CD3ζ and Lat in live T cells on bilayers by dcFCCS.
Figure 5: Kinetics of the correlation between CD3ζ and Lat themselves and each other in T cells on nonactivating and activating bilayers.

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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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Author notes

  1. Björn F Lillemeier & Martin B Forstner

    Present address: Present addresses: Nomis Center for Immunobiology and Microbial Pathogenesis, Waitt Advanced Biophotonics Center, The Salk Institute, La Jolla, California, USA (B.F.L.), and Department of Physics, Syracuse Biomaterials Institute, Syracuse University, Syracuse, New York, USA (M.B.F.).,

Authors and Affiliations

  1. Department of Microbiology & Immunology, Howard Hughes Medical Institute, Stanford University School of Medicine, Stanford, California, USA

    Björn F Lillemeier, Manuel A Mörtelmaier, Johannes B Huppa & Mark M Davis

  2. Department of Chemistry, Physical Bioscience and Materials Sciences Divisions, Howard Hughes Medical Institute, University of California Berkeley, Lawrence Berkeley National Laboratory, Berkeley, California, USA

    Martin B Forstner & Jay T Groves

Authors
  1. Björn F Lillemeier
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  2. Manuel A Mörtelmaier
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  6. Mark M Davis
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Contributions

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.

Corresponding author

Correspondence to Mark M Davis.

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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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