title: Monitoring Ligand-Activated Protein-Protein Interactions Using Bioluminescent Resonance Energy Transfer (BRET) Assay.
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.
authors: Department of Oncology, University of Wisconsin-Madison, 7459 WIMR II, 1111 Highland Avenue, Madison, WI, 53705-2275, USA.
authors: Carlos
authors: C
authors: Coriano
authors: Department of Experimental Radiation Oncology, The University of Texas MD Anderson Cancer Center, Houston, TX, 77030, USA.
authors: Emily
authors: E
authors: Powell
authors: Department of Oncology, University of Wisconsin-Madison, 7459 WIMR II, 1111 Highland Avenue, Madison, WI, 53705-2275, USA. wxu@oncology.wisc.edu.
authors: Wei
authors: W
authors: Xu
mesh_terms: D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism; D060807:Bioluminescence Resonance Energy Transfer Techniques; D052244:Endocrine Disruptors / Q000633:toxicity*; D047628:Estrogen Receptor alpha / Q000235:genetics* / Q000378:metabolism; D047629:Estrogen Receptor beta / Q000235:genetics* / Q000378:metabolism; D005786:Gene Expression Regulation / Q000187:drug effects*; D057809:HEK293 Cells; D057166:High-Throughput Screening Assays; D006801:Humans; D008156:Luciferases / Q000235:genetics / Q000378:metabolism; D008164:Luminescent Proteins / Q000235:genetics / Q000378:metabolism; D010957:Plasmids / Q000737:chemistry / Q000378:metabolism; D025941:Protein Interaction Mapping; D055503:Protein Multimerization / Q000187:drug effects; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism
chemical_list: D001426:Bacterial Proteins; C506487:ESR1 protein, human; D052244:Endocrine Disruptors; D047628:Estrogen Receptor alpha; D047629:Estrogen Receptor beta; D008164:Luminescent Proteins; D011993:Recombinant Fusion Proteins; C054562:yellow fluorescent protein, Bacteria; D008156:Luciferases
keywords: Bioluminescent Resonance Energy Transfer (BRET); Imaging assay; Protein–protein interactions; Screening assay