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Fluorescence Correlation Spectroscopy to Examine Protein–Lipid Interactions in Membranes

  • Viktoria Betaneli
  • Jonas Mücksch
  • Petra SchwilleEmail author
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 2003)

Abstract

Fluorescence correlation spectroscopy (FCS) is a versatile technique to study membrane dynamics and protein–lipid interactions. It can provide information about diffusion coefficients, concentrations, and molecular interactions of proteins and lipids in the membrane. These parameters allow for the determination of protein partitioning into different lipid environments, the identification of lipid domains, and the detection of lipid–protein complexes on the membrane. During the last decades, FCS studies were successfully performed on model membrane systems as also on living cells, to characterize protein–lipid interactions. Recent developments of the method described here improved quantitative measurements on membranes and decreased the number of potential artifacts. The aim of this chapter is to provide the reader with the necessary information and some practical guidelines to perform FCS studies on artificial and cellular membranes.

Key words

Fluorescence correlation spectroscopy FCS Cross-correlation FCCS Diffusion Model membranes Giant unilamellar vesicles Protein–lipid interactions Lipid rafts 

Notes

Acknowledgments

We thank Dr. Fabian Heinemann for the critical reading of the manuscript, Alena Khmelinskaia for discussions on the lipid monolayer protocol, and Dr. Erdinc Sezgin for the help with the protocol describing the preparation of living cells for FCS. This work was supported by the Dresden International Graduate School for Biomedicine and Bioengineering. J.M. acknowledges support from the International Max Planck Research School acknowledges support from the excellence cluster Nanosystems Initiative Munich and the International Max Planck Research School for Molecular Life Sciences.

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© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  • Viktoria Betaneli
    • 1
  • Jonas Mücksch
    • 2
  • Petra Schwille
    • 2
    Email author
  1. 1.Medical Faculty “Carl Gustav Carus”, Institute of Physiological ChemistryTechnische Universität DresdenDresdenGermany
  2. 2.Max Planck Institute of BiochemistryMartinsriedGermany

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