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. 2016 Aug 1:85:29.17.1-29.17.13.
doi: 10.1002/cpps.10.

Correlative Förster Resonance Electron Transfer-Proximity Ligation Assay (FRET-PLA) Technique for Studying Interactions Involving Membrane Proteins

Daniel Ivanusic  1 , Joachim Denner  1 , Norbert Bannert  1
Affiliations

Correlative Förster Resonance Electron Transfer-Proximity Ligation Assay (FRET-PLA) Technique for Studying Interactions Involving Membrane Proteins

Daniel Ivanusic et al. Curr Protoc Protein Sci. .

Abstract

This unit provides a guide and detailed protocol for studying membrane protein-protein interactions (PPI) using the acceptor-sensitized Förster resonance electron transfer (FRET) method in combination with the proximity ligation assay (PLA). The protocol in this unit is focused on the preparation of FRET-PLA samples and the detection of correlative FRET/PLA signals as well as on the analysis of FRET-PLA data and interpretation of correlative results when using cyan fluorescent protein (CFP) as a FRET donor and yellow fluorescent protein (YFP) as a FRET acceptor. The correlative application of FRET and PLA combines two powerful tools for monitoring PPI, yielding results that are more reliable than with either technique alone. © 2016 by John Wiley & Sons, Inc.

Keywords: FRET; NFRET; PLA; membrane proteins; protein-protein interaction; proximity ligation assay; sensitized emission.

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