气候变化领域集成服务门户(CCPortal): BRET-based effector membrane translocation assay monitors GPCR-promoted and endocytosis-mediated Gqactivation at early endosomes

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DOI	10.1073/pnas.2025846118
	BRET-based effector membrane translocation assay monitors GPCR-promoted and endocytosis-mediated Gqactivation at early endosomes
	Wright S.C.; Lukasheva V.; Gouill C.L.; Kobayashi H.; Breton B.; Mailhot-Larouche S.; Blondel-Tepaz E.; Vieira N.A.; Costa-Neto C.; Héroux M.; Lambert N.A.; Parreiras-E-Silva L.T.; Bouvier M.
发表日期	2021
ISSN	0027-8424
卷号	118 期号:20
英文摘要	G protein-coupled receptors (GPCRs) are gatekeepers of cellular homeostasis and the targets of a large proportion of drugs. In addition to their signaling activity at the plasma membrane, it has been proposed that their actions may result from translocation and activation of G proteins at endomembranes-namely endosomes. This could have a significant impact on our understanding of how signals from GPCR-targeting drugs are propagated within the cell. However, little is known about the mechanisms that drive G protein movement and activation in subcellular compartments. Using bioluminescence resonance energy transfer (BRET)-based effector membrane translocation assays, we dissected the mechanisms underlying endosomal Gqtrafficking and activity following activation of Gq-coupled receptors, including the angiotensin II type 1, bradykinin B2, oxytocin, thromboxane A2alpha isoform, and muscarinic acetylcholine M3receptors. Our data reveal that GPCR-promoted activation of Gqat the plasma membrane induces its translocation to endosomes independently of β-arrestin engagement and receptor endocytosis. In contrast, Gq activity at endosomes was found to rely on both receptor endocytosisdependent and -independent mechanisms. In addition to shedding light on the molecular processes controlling subcellular Gqsignaling, our study provides a set of tools that will be generally applicable to the study of G protein translocation and activation at endosomes and other subcellular organelles, as well as the contribution of signal propagation to drug action. © 2021 National Academy of Sciences. All rights reserved.
英文关键词	Arrestin; Endosomal signaling; GPCR; Gq/11
语种	英语
scopus关键词	angiotensin 1 receptor; angiotensin II; beta arrestin; bradykinin B2 receptor; G protein coupled receptor; muscarinic M3 receptor; oxytocin; thromboxane A2; acidification; Article; bioluminescence resonance energy transfer; cell activation; cell membrane; cell organelle; endocytosis; endosome; intracellular space; membrane transport; protein function; protein localization; protein transport; signal transduction
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/238919
作者单位	Department of Biochemistry and Molecular Medicine, Université de Montreál, Montreál, QC H3T 1J4, Canada; Institute for Research in Immunology and Cancer, Université de Montreál, Montreál, QC H3T 1J4, Canada; Molecular Biology Program, Université de Montreál, Montreál, QC H3T 1J4, Canada; Department of Biochemistry and Immunology, Ribeirão Preto Medical School, University of São PauloSP 14049-900, Brazil; Department of Pharmacology and Toxicology, Medical College of Georgia, Augusta University, Augusta, GA 30912, United States
推荐引用方式 GB/T 7714	Wright S.C.,Lukasheva V.,Gouill C.L.,et al. BRET-based effector membrane translocation assay monitors GPCR-promoted and endocytosis-mediated Gqactivation at early endosomes[J],2021,118(20).
APA	Wright S.C..,Lukasheva V..,Gouill C.L..,Kobayashi H..,Breton B..,...&Bouvier M..(2021).BRET-based effector membrane translocation assay monitors GPCR-promoted and endocytosis-mediated Gqactivation at early endosomes.Proceedings of the National Academy of Sciences of the United States of America,118(20).
MLA	Wright S.C.,et al."BRET-based effector membrane translocation assay monitors GPCR-promoted and endocytosis-mediated Gqactivation at early endosomes".Proceedings of the National Academy of Sciences of the United States of America 118.20(2021).