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

Kevin Pfleger, Jasmin Dromey, Matthew Dalrymple, Esther Lim, W.G. Thomas, Karin Eidne

Research output: Contribution to journalArticle

79 Citations (Scopus)

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/beta-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. (c) 2006 Elsevier Inc. All rights reserved.
Original languageEnglish
Pages (from-to)1664-1670
JournalCellular Signalling
Volume18
Issue number10
DOIs
Publication statusPublished - 2006

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