Extended bioluminescence resonance energy transfer (eBRET) for monitoring prolonged protein–protein interactions in live cells

Pfleger, Kevin D. G., Dromey, Jasmin R., Dalrymple, Matthew B., Lim, Esther M. L., Thomas, Walter G. and Eidne, Karin A. (2006) Extended bioluminescence resonance energy transfer (eBRET) for monitoring prolonged protein–protein interactions in live cells. Cellular Signalling, 18 10: 1664-1670. doi:10.1016/j.cellsig.2006.01.004


Author Pfleger, Kevin D. G.
Dromey, Jasmin R.
Dalrymple, Matthew B.
Lim, Esther M. L.
Thomas, Walter G.
Eidne, Karin A.
Title Extended bioluminescence resonance energy transfer (eBRET) for monitoring prolonged protein–protein interactions in live cells
Journal name Cellular Signalling   Check publisher's open access policy
ISSN 0898-6568
Publication date 2006-10-01
Sub-type Article (original research)
DOI 10.1016/j.cellsig.2006.01.004
Volume 18
Issue 10
Start page 1664
End page 1670
Total pages 7
Place of publication Oxford
Publisher Elsevier/Pergamon
Language eng
Subject 060110 Receptors and Membrane Biology
060111 Signal Transduction
060199 Biochemistry and Cell Biology not elsewhere classified
060108 Protein Trafficking
060602 Animal Physiology - Cell
110201 Cardiology (incl. Cardiovascular Diseases)
110903 Central Nervous System
Abstract Bioluminescence resonance energy transfer (BRET) is an increasingly popular technique for studying protein–protein interactions in live cells. It is particularly suitable for real-time monitoring of such interactions, however, the timescale over which assays can be carried out is currently relatively short (minutes) due to substrate instability. We present a new derivation of the BRET technology, termed ‘extended BRET’ (eBRET), which now enables protein–protein interactions to be monitored in real-time for many hours. This capability has significant benefits for investigating cellular function over extended timescales, as we have illustrated using the agonist-induced G-protein coupled receptor/β-arrestin interaction. The potential for studying the modulation of such interactions by agonists, antagonists, inhibitors, dominant negative mutants and co-expressed accessory proteins is substantial. Furthermore, the advantages of eBRET have important implications for the development of high-throughput BRET screening systems, an ever-expanding area of interest for the pharmaceutical industry.
Keyword Bioluminescence resonance energy transfer
Protein–protein interaction
BRET
G-protein coupled receptor
β-arrestin
Q-Index Code C1

Document type: Journal Article
Sub-type: Article (original research)
Collections: Excellence in Research Australia (ERA) - Collection
School of Biomedical Sciences Publications
 
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