Hdl Handle:
http://hdl.handle.net/10675.2/735
Title:
Internalization Dissociates b2-Adrenergic Receptors
Authors:
Lan, Tien-Hung; Kuravi, Sudhakiranmayi; Lambert, Nevin A.
Abstract:
G protein-coupled receptors (GPCRs) self-associate as dimers or higher-order oligomers in living cells. The stability of associated GPCRs has not been extensively studied, but it is generally thought that these receptors move between the plasma membrane and intracellular compartments as intact dimers or oligomers. Here we show that b2-adrenergic receptors (b2ARs) that self-associate at the plasma membrane can dissociate during agonist-induced internalization. We use bioluminescence-resonance energy transfer (BRET) to monitor movement of β2ARs between subcellular compartments. BRET between b2ARs and plasma membrane markers decreases in response to agonist activation, while at the same time BRET between b2ARs and endosome markers increases. Energy transfer between b2ARs is decreased in a similar manner if either the donor- or acceptor-labeled receptor is mutated to impair agonist binding and internalization. These changes take place over the course of 30 minutes, persist after agonist is removed, and are sensitive to several inhibitors of arrestin- and clathrin-mediated endocytosis. The magnitude of the decrease in BRET between donor- and acceptor-labeled b2ARs suggests that at least half of the receptors that contribute to the BRET signal are physically segregated by internalization. These results are consistent with the possibility that b2ARs associate transiently with each other in the plasma membrane, or that b2AR dimers or oligomers are actively disrupted during internalization.
Citation:
PLoS One. 2011 Feb 22; 6(2):e17361
Issue Date:
22-Feb-2011
URI:
http://hdl.handle.net/10675.2/735
DOI:
10.1371/journal.pone.0017361
PubMed ID:
21364942
PubMed Central ID:
PMC3043075
Type:
Article
ISSN:
1932-6203
Appears in Collections:
Department of Pharmacology and Toxicology: Faculty Research and Presentations

Full metadata record

DC FieldValue Language
dc.contributor.authorLan, Tien-Hungen_US
dc.contributor.authorKuravi, Sudhakiranmayien_US
dc.contributor.authorLambert, Nevin A.en_US
dc.date.accessioned2012-10-26T20:27:16Z-
dc.date.available2012-10-26T20:27:16Z-
dc.date.issued2011-02-22en_US
dc.identifier.citationPLoS One. 2011 Feb 22; 6(2):e17361en_US
dc.identifier.issn1932-6203en_US
dc.identifier.pmid21364942en_US
dc.identifier.doi10.1371/journal.pone.0017361en_US
dc.identifier.urihttp://hdl.handle.net/10675.2/735-
dc.description.abstractG protein-coupled receptors (GPCRs) self-associate as dimers or higher-order oligomers in living cells. The stability of associated GPCRs has not been extensively studied, but it is generally thought that these receptors move between the plasma membrane and intracellular compartments as intact dimers or oligomers. Here we show that b2-adrenergic receptors (b2ARs) that self-associate at the plasma membrane can dissociate during agonist-induced internalization. We use bioluminescence-resonance energy transfer (BRET) to monitor movement of β2ARs between subcellular compartments. BRET between b2ARs and plasma membrane markers decreases in response to agonist activation, while at the same time BRET between b2ARs and endosome markers increases. Energy transfer between b2ARs is decreased in a similar manner if either the donor- or acceptor-labeled receptor is mutated to impair agonist binding and internalization. These changes take place over the course of 30 minutes, persist after agonist is removed, and are sensitive to several inhibitors of arrestin- and clathrin-mediated endocytosis. The magnitude of the decrease in BRET between donor- and acceptor-labeled b2ARs suggests that at least half of the receptors that contribute to the BRET signal are physically segregated by internalization. These results are consistent with the possibility that b2ARs associate transiently with each other in the plasma membrane, or that b2AR dimers or oligomers are actively disrupted during internalization.en_US
dc.rightsLan et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.en_US
dc.subjectResearch Articleen_US
dc.subjectBiologyen_US
dc.subjectBiochemistryen_US
dc.subjectProteinsen_US
dc.subjectLuminescent Proteinsen_US
dc.subjectTransmembrane Proteinsen_US
dc.subjectChemical Biologyen_US
dc.subjectBiophysicsen_US
dc.subjectMacromolecular Assembliesen_US
dc.subjectMolecular Cell Biologyen_US
dc.subjectSignal Transductionen_US
dc.subjectMembrane Receptor Signalingen_US
dc.subjectHormone Receptor Signalingen_US
dc.subjectNeurotransmitter Receptor Signalingen_US
dc.subjectSignaling in Cellular Processesen_US
dc.subjectG-Protein Signalingen_US
dc.subjectChemistryen_US
dc.subjectPhysical Chemistryen_US
dc.subjectEnergy Transferen_US
dc.titleInternalization Dissociates b2-Adrenergic Receptorsen_US
dc.typeArticleen_US
dc.identifier.pmcidPMC3043075en_US
dc.contributor.corporatenameDepartment of Pharmacology and Toxicology-

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