Monitoring Transmembrane and Peripheral Membrane Protein Interactions by Förster Resonance Energy Transfer Using Fluorescence Lifetime Imaging Microscopy

Janhavi Nagwekar, Caterina Di Ciano-Oliveira, Gregory D. Fairn

Research output: Chapter in Book/Report/Conference proceedingChapter

3 Citations (Scopus)

Abstract

Caveolae are bulb-shaped invaginations of the plasma membrane that are enriched in specific lipids including cholesterol, phosphatidylserine and sphingolipids. Caveolae have many described cellular roles and functions, including endocytic transport, transcytosis, mechanosensing, and serving as a buffer against plasmalemmal stress. Caveola are formed through interactions between integral membrane proteins (Caveolin) and a cavin family of peripheral proteins (Cavins). Nearly half of the human proteome resides within or at the surface of membranes. Studying protein–protein interactions, especially of transmembrane domain containing proteins can be challenging. Fortunately, sophisticated biophysical methods allow for the monitoring of protein interactions in intact cells. Here, we describe the principles of Förster resonance energy transfer, fluorescence lifetime, and how their properties can be used to assess protein–protein interactions. Additionally, we discuss and demonstrate how fluorescence lifetime can be monitored microscopically thereby providing caveolin–cavin interaction data from living cells.

Original languageEnglish
Title of host publicationMethods in Molecular Biology
PublisherHumana Press Inc.
Pages77-90
Number of pages14
DOIs
Publication statusPublished - 2022

Publication series

NameMethods in Molecular Biology
Volume2440
ISSN (Print)1064-3745
ISSN (Electronic)1940-6029

Bibliographical note

Funding Information:
Original work in the laboratory of G.D.F. is supported by the Canadian Institutes of Health Research (Grants: PJT166010 and PJT165968) and the Natural Sciences and Engineering Research Council of Canada. Figures 1–3 were generated using BioRender. com. Conflict of Interest: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.

Publisher Copyright:
© 2022, The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature.

ASJC Scopus Subject Areas

  • Molecular Biology
  • Genetics

PubMed: MeSH publication types

  • Journal Article
  • Research Support, Non-U.S. Gov't

Fingerprint

Dive into the research topics of 'Monitoring Transmembrane and Peripheral Membrane Protein Interactions by Förster Resonance Energy Transfer Using Fluorescence Lifetime Imaging Microscopy'. Together they form a unique fingerprint.

Cite this