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Caveolae pp 71-80 | Cite as

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

  • Adriana M. Zimnicka
  • Zhenlong Chen
  • Peter T. Toth
  • Richard D. MinshallEmail author
Protocol
Part of the Methods in Molecular Biology book series (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.

Key words

Live-cell FRET Cav-1-CFP Cav-1-YFP Caveolae Conformation change 

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Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2020

Authors and Affiliations

  • Adriana M. Zimnicka
    • 1
  • Zhenlong Chen
    • 2
  • Peter T. Toth
    • 1
    • 3
  • Richard D. Minshall
    • 1
    • 2
    Email author
  1. 1.Department of PharmacologyUniversity of Illinois at ChicagoChicagoUSA
  2. 2.Department of and AnesthesiologyUniversity of Illinois at ChicagoChicagoUSA
  3. 3.Research Resources Center Fluorescence Imaging CoreUniversity of Illinois at ChicagoChicagoUSA

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