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Live-Cell FRET Imaging of Phosphorylation-Dependent Caveolin-1 Switch

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Caveolae

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2169))

Abstract

The detection of dynamic conformational changes in proteins in live cells is challenging. Live-cell FRET (Förster Resonance Energy Transfer) is an example of a noninvasive technique that can be used to achieve this goal at nanometer resolution. FRET-based assays are dependent on the presence of fluorescent probes, such as CFP- and YFP-conjugated protein pairs. Here, we describe an experimental protocol in which live-cell FRET was used to measure conformational changes in caveolin-1 (Cav-1) oligomers on the surface of plasmalemma vesicles, or caveolae.

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Author information

Authors and Affiliations

  1. Department of Pharmacology, University of Illinois at Chicago, Chicago, IL, USA

    Adriana M. Zimnicka, Peter T. Toth & Richard D. Minshall

  2. Department of and Anesthesiology, University of Illinois at Chicago, Chicago, IL, USA

    Zhenlong Chen & Richard D. Minshall

  3. Research Resources Center Fluorescence Imaging Core, University of Illinois at Chicago, Chicago, IL, USA

    Peter T. Toth

Authors
  1. Adriana M. Zimnicka
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  2. Zhenlong Chen
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  3. Peter T. Toth
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  4. Richard D. Minshall
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Corresponding author

Correspondence to Richard D. Minshall .

Editor information

Editors and Affiliations

  1. Institut Curie – Centre de Recherche, PSL Research University, Membrane Mechanics and Dynamics of Intracellular Signaling Laboratory, Paris, France

    Cedric M. Blouin

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Zimnicka, A.M., Chen, Z., Toth, P.T., Minshall, R.D. (2020). Live-Cell FRET Imaging of Phosphorylation-Dependent Caveolin-1 Switch. In: Blouin, C. (eds) Caveolae. Methods in Molecular Biology, vol 2169. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-0732-9_7

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  • DOI: https://doi.org/10.1007/978-1-0716-0732-9_7

  • Published:

  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-0731-2

  • Online ISBN: 978-1-0716-0732-9

  • eBook Packages: Springer Protocols

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