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TNF Receptor Membrane Dynamics Studied with Fluorescence Microscopy and Spectroscopy

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Fluorescent Methods to Study Biological Membranes

Part of the book series: Springer Series on Fluorescence ((SS FLUOR,volume 13))

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Abstract

Sensitive fluorescence techniques opened novel opportunities to study the function and interaction of proteins in living cells. Here, we review the contribution of fluorescence correlation spectroscopy (FCS) and single-molecule tracking to study the dynamics of TNF receptor 1 and 2 (TNFR1/2). Although these techniques greatly differ, both report a similar behavior of TNF receptor (TNFR) species in Hela cells under different experimental conditions. FCS as well as single-molecule tracking revealed an increase of the diffusion coefficient of TNFR1 after treating cells with methyl-cyclodextrin.

In addition, FCS studies of the activation of TNFR1 showed that ligand binding hardly affects its diffusion coefficient.

In contrast, unstimulated TNFR2 was observed to diffuse faster than TNFR1, whereas ligand stimulation of TNFR2 decreases the diffusion coefficient.

In conclusion, the results indicate that the two TNFRs compartmentalize in distinct domains of the plasma membrane most likely determined by the respective transmembrane domains and/or transmembrane domain near regions.

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Author information

Authors and Affiliations

  1. Institute of Cell Biology and Immunology, University of Stuttgart, Allmandring 31, D-70569, Stuttgart, Germany

    Felix Neugart

  2. Leica Microsystems CMS GmbH, Am Friedensplatz 3, D-68165, Mannheim, Germany

    Felix Neugart

  3. Cell Biology, University of Bielefeld, Universitätsstr. 25, D-33501, Bielefeld, Germany

    Darius Widera & Christian Kaltschmidt

  4. Molecular Neurobiology, University of Bielefeld, Universitätsstr. 25, D-33501, Bielefeld, Germany

    Barbara Kaltschmidt

  5. Department of Biotechnology & Biophysics, Julius-Maximilians-Universität, Am Hubland, D-97074, Würzburg, Germany

    Mike Heilemann

Authors
  1. Felix Neugart
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  2. Darius Widera
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  3. Barbara Kaltschmidt
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  4. Christian Kaltschmidt
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  5. Mike Heilemann
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Corresponding authors

Correspondence to Felix Neugart , Darius Widera or Mike Heilemann .

Editor information

Editors and Affiliations

  1. Faculté de Pharmacie, Laboratoire de Biophotonique, Université de Strasbourg, route du Rhin 74, Illkirch, 67401, France

    Yves Mély

  2. Faculté de Pharmacie, Laboratoire de Biophotonique et, Université de Strasbourg, route du Rhin 74, Illkirch, 67401, France

    Guy Duportail

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© 2012 Springer-Verlag Berlin Heidelberg

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Neugart, F., Widera, D., Kaltschmidt, B., Kaltschmidt, C., Heilemann, M. (2012). TNF Receptor Membrane Dynamics Studied with Fluorescence Microscopy and Spectroscopy. In: Mély, Y., Duportail, G. (eds) Fluorescent Methods to Study Biological Membranes. Springer Series on Fluorescence, vol 13. Springer, Berlin, Heidelberg. https://doi.org/10.1007/4243_2012_60

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