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DOI	10.1073/pnas.1716224115
	Monitoring ligand-dependent assembly of receptor ternary complexes in live cells by BRETFect
	Cotnoir-White D.; Ezzy M.E.; Boulay P.-L.; Rozendaal M.; Bouvier M.; Gagnon E.; Mader S.
发表日期	2018
ISSN	0027-8424
起始页码	E2653
结束页码	E2662
卷号	115 期号:11
英文摘要	There is currently an unmet need for versatile techniques to monitor the assembly and dynamics of ternary complexes in live cells. Here we describe bioluminescence resonance energy transfer with fluorescence enhancement by combined transfer (BRETFect), a high-throughput technique that enables robust spectrometric detection of ternary protein complexes based on increased energy transfer from a luciferase to a fluorescent acceptor in the presence of a fluorescent intermediate. Its unique donor–intermediate–acceptor relay system is designed so that the acceptor can receive energy either directly from the donor or indirectly via the intermediate in a combined transfer, taking advantage of the entire luciferase emission spectrum. BRETFect was used to study the ligand-dependent cofactor interaction properties of the estrogen receptors ERα and ERβ, which form homo- or heterodimers whose distinctive regulatory properties are difficult to dissect using traditional methods. BRETFect uncovered the relative capacities of hetero- vs. homodimers to recruit receptor-specific cofactors and regulatory proteins, and to interact with common cofactors in the presence of receptor-specific ligands. BRETFect was also used to follow the assembly of ternary complexes between the V2R vasopressin receptor and two different intracellular effectors, illustrating its use for dissection of ternary protein–protein interactions engaged by G protein-coupled receptors. Our results indicate that BRETFect represents a powerful and versatile technique to monitor the dynamics of ternary interactions within multimeric complexes in live cells. © 2018 National Academy of Sciences. All Rights Reserved.
英文关键词	Bioluminescence resonance energy transfer; G protein-coupled receptors; Heterodimers; Nuclear receptors; Ternary complexes
语种	英语
scopus关键词	1,3,5 tris(4 hydroxyphenyl) 4 propylpyrazole; afimoxifene; diarylpropionitrile; estrogen receptor alpha; estrogen receptor beta; fulvestrant; G protein coupled receptor; heterodimer; homodimer; luciferase; vasopressin V2 receptor; cell receptor; G protein coupled receptor; photoprotein; ternary complex factor; Article; bioluminescence resonance energy transfer with fluorescence enhancement by combined transfer; complex formation; controlled study; dimerization; drug potency; drug selectivity; embryo; energy transfer; human; human cell; ligand binding; priority journal; protein analysis; protein protein interaction; sumoylation; chemistry; cytology; fluorescence resonance energy transfer; HEK293 cell line; metabolism; procedures; Cytological Techniques; Fluorescence Resonance Energy Transfer; HEK293 Cells; Humans; Luminescent Proteins; Receptors, Cytoplasmic and Nuclear; Receptors, G-Protein-Coupled; Ternary Complex Factors
来源期刊	Proceedings of the National Academy of Sciences of the United States of America
文献类型	期刊论文
条目标识符	http://gcip.llas.ac.cn/handle/2XKMVOVA/160556
作者单位	Cotnoir-White, D., Institute for Research in Immunology and Cancer, Université de Montréal, Montréal, QC H3T 1J4, Canada; Ezzy, M.E., Institute for Research in Immunology and Cancer, Université de Montréal, Montréal, QC H3T 1J4, Canada; Boulay, P.-L., Institute for Research in Immunology and Cancer, Université de Montréal, Montréal, QC H3T 1J4, Canada; Rozendaal, M., Institute for Research in Immunology and Cancer, Université de Montréal, Montréal, QC H3T 1J4, Canada; Bouvier, M., Institute for Research in Immunology and Cancer, Université de Montréal, Montréal, QC H3T 1J4, Canada, Département de Biochimie et Médecine Moléculaire, Université de Montréal, Montréal, QC H3T 1J4, Canada; Gagnon, E., Institute for Research in Immunology and Cancer, Université de Montréal, Montréal, QC H3T 1J4, Canada, Département de Microbiologie, Infectiologie et Immunologie, Université de Montréal, Montréal, QC H3T 1J4, Canada; Mader, S., Institute for Research in Immunology and Cancer, Université de Montréal, Montré...
推荐引用方式 GB/T 7714	Cotnoir-White D.,Ezzy M.E.,Boulay P.-L.,et al. Monitoring ligand-dependent assembly of receptor ternary complexes in live cells by BRETFect[J],2018,115(11).
APA	Cotnoir-White D..,Ezzy M.E..,Boulay P.-L..,Rozendaal M..,Bouvier M..,...&Mader S..(2018).Monitoring ligand-dependent assembly of receptor ternary complexes in live cells by BRETFect.Proceedings of the National Academy of Sciences of the United States of America,115(11).
MLA	Cotnoir-White D.,et al."Monitoring ligand-dependent assembly of receptor ternary complexes in live cells by BRETFect".Proceedings of the National Academy of Sciences of the United States of America 115.11(2018).