title: Correlative Förster Resonance Electron Transfer-Proximity Ligation Assay (FRET-PLA) Technique for Studying Interactions Involving Membrane Proteins.
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.
authors.affiliation: Robert Koch Institute, HIV and Other Retroviruses, Berlin, Germany.
authors.forename: Daniel
authors.initials: D
authors.lastname: Ivanusic
authors.affiliation: Robert Koch Institute, HIV and Other Retroviruses, Berlin, Germany.
authors.forename: Joachim
authors.initials: J
authors.lastname: Denner
authors.affiliation: Robert Koch Institute, HIV and Other Retroviruses, Berlin, Germany.
authors.forename: Norbert
authors.initials: N
authors.lastname: Bannert
mesh_terms: D001426:Bacterial Proteins / Q000737:chemistry; D031541:Fluorescence Resonance Energy Transfer / Q000379:methods*; D049452:Green Fluorescent Proteins / Q000737:chemistry; D057809:HEK293 Cells; D006801:Humans; D008026:Ligation; D008164:Luminescent Proteins / Q000737:chemistry; D008565:Membrane Proteins / Q000737:chemistry*; D008856:Microscopy, Fluorescence; D011485:Protein Binding; D011487:Protein Conformation; D025941:Protein Interaction Mapping / Q000379:methods*; D011499:Protein Processing, Post-Translational
chemical_list: D001426:Bacterial Proteins; C485182:Cyan Fluorescent Protein; D008164:Luminescent Proteins; D008565:Membrane Proteins; C054562:yellow fluorescent protein, Bacteria; D049452:Green Fluorescent Proteins
keywords: FRET; NFRET; PLA; membrane proteins; protein-protein interaction; proximity ligation assay; sensitized emission