期刊
CELLULAR SIGNALLING
卷 18, 期 10, 页码 1664-1670出版社
ELSEVIER SCIENCE INC
DOI: 10.1016/j.cellsig.2006.01.004
关键词
bioluminescence resonance energy transfer; BRET; protein-protein interaction; G-protein coupled receptor; beta-arrestin
类别
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.
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