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Monitoring Ligand-Activated Protein–Protein Interactions Using Bioluminescent Resonance Energy Transfer (BRET) Assay

  • Carlos Coriano
  • Emily Powell
  • Wei XuEmail author
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1473)

Abstract

The bioluminescent resonance energy transfer (BRET) assay has been extensively used in cell-based and in vivo imaging systems for detecting protein–protein interactions in the native environment of living cells. These protein–protein interactions are essential for the functional response of many signaling pathways to environmental chemicals. BRET has been used as a toxicological tool for identifying chemicals that either induce or inhibit these protein–protein interactions. This chapter focuses on describing the toxicological applications of BRET and its optimization as a high-throughput detection system in live cells. Here we review the construction of BRET fusion proteins, describe the BRET methodology, and outline strategies to overcome obstacles that may arise. Furthermore, we describe the advantage of BRET over other resonance energy transfer methods for monitoring protein–protein interactions.

Key words

Bioluminescent Resonance Energy Transfer (BRET) Protein–protein interactions Screening assay Imaging assay 

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Copyright information

© Springer Science+Business Media New York 2016

Authors and Affiliations

  1. 1.Department of OncologyUniversity of Wisconsin-MadisonMadisonUSA
  2. 2.Department of Experimental Radiation OncologyThe University of Texas MD Anderson Cancer CenterHoustonUSA

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