title: Allosteric modulation of M1 muscarinic acetylcholine receptor internalization and subcellular trafficking.
abstract: Allosteric modulators are an attractive approach to achieve receptor subtype-selective targeting of G protein-coupled receptors. Benzyl quinolone carboxylic acid (BQCA) is an unprecedented example of a highly selective positive allosteric modulator of the M1 muscarinic acetylcholine receptor (mAChR). However, despite favorable pharmacological characteristics of BQCA in vitro and in vivo, there is limited evidence of the impact of allosteric modulation on receptor regulatory mechanisms such as β-arrestin recruitment or receptor internalization and endocytic trafficking. In the present study we investigated the impact of BQCA on M1 mAChR regulation. We show that BQCA potentiates agonist-induced β-arrestin recruitment to M1 mAChRs. Using a bioluminescence resonance energy transfer approach to monitor intracellular trafficking of M1 mAChRs, we show that once internalized, M1 mAChRs traffic to early endosomes, recycling endosomes and late endosomes. We also show that BQCA potentiates agonist-induced subcellular trafficking. M1 mAChR internalization is both β-arrestin and G protein-dependent, with the third intracellular loop playing an important role in the dynamics of β-arrestin recruitment. As the global effect of receptor activation ultimately depends on the levels of receptor expression at the cell surface, these results illustrate the need to extend the characterization of novel allosteric modulators of G protein-coupled receptors to encapsulate the consequences of chronic exposure to this family of ligands.
authors.affiliation: From the Drug Discovery Biology, Monash Institute of Pharmaceutical Sciences, Faculty of Pharmacy and Pharmaceutical Sciences, Monash University, Parkville, Melbourne, Victoria 3052, Australia and.
authors.forename: Holly R
authors.initials: HR
authors.lastname: Yeatman
authors.affiliation: From the Drug Discovery Biology, Monash Institute of Pharmaceutical Sciences, Faculty of Pharmacy and Pharmaceutical Sciences, Monash University, Parkville, Melbourne, Victoria 3052, Australia and.
authors.forename: J Robert
authors.initials: JR
authors.lastname: Lane
authors.affiliation: From the Drug Discovery Biology, Monash Institute of Pharmaceutical Sciences, Faculty of Pharmacy and Pharmaceutical Sciences, Monash University, Parkville, Melbourne, Victoria 3052, Australia and.
authors.forename: Kwok Ho Christopher
authors.initials: KH
authors.lastname: Choy
authors.affiliation: Department of Pharmacology and Toxicology, Georgia Regents University, Augusta, Georgia 30912.
authors.forename: Nevin A
authors.initials: NA
authors.lastname: Lambert
authors.affiliation: From the Drug Discovery Biology, Monash Institute of Pharmaceutical Sciences, Faculty of Pharmacy and Pharmaceutical Sciences, Monash University, Parkville, Melbourne, Victoria 3052, Australia and.
authors.forename: Patrick M
authors.initials: PM
authors.lastname: Sexton
authors.affiliation: From the Drug Discovery Biology, Monash Institute of Pharmaceutical Sciences, Faculty of Pharmacy and Pharmaceutical Sciences, Monash University, Parkville, Melbourne, Victoria 3052, Australia and arthur.christopoulos@monash.edu.
authors.forename: Arthur
authors.initials: A
authors.lastname: Christopoulos
authors.affiliation: From the Drug Discovery Biology, Monash Institute of Pharmaceutical Sciences, Faculty of Pharmacy and Pharmaceutical Sciences, Monash University, Parkville, Melbourne, Victoria 3052, Australia and meri.canals@monash.edu.
authors.forename: Meritxell
authors.initials: M
authors.lastname: Canals
mesh_terms: D000494:Allosteric Regulation / Q000502:physiology; D000818:Animals; D019390:Arrestins / Q000378:metabolism; D016466:CHO Cells; D055884:Chemical Hazard Release; D003412:Cricetulus; D004705:Endocytosis / Q000502:physiology*; D011992:Endosomes / Q000378:metabolism*; D054769:G-Protein-Coupled Receptor Kinase 2 / Q000378:metabolism; D043802:GTP-Binding Protein alpha Subunits, Gq-G11 / Q000378:metabolism; D057809:HEK293 Cells; D006801:Humans; D008024:Ligands; D021381:Protein Transport / Q000502:physiology; D043643:Receptor, Muscarinic M1 / Q000737:chemistry* / Q000378:metabolism*; D043562:Receptors, G-Protein-Coupled / Q000378:metabolism; D015398:Signal Transduction / Q000502:physiology; D000071557:beta-Arrestins
chemical_list: D019390:Arrestins; D008024:Ligands; D043643:Receptor, Muscarinic M1; D043562:Receptors, G-Protein-Coupled; D000071557:beta-Arrestins; C496567:GRK2 protein, human; D054769:G-Protein-Coupled Receptor Kinase 2; D043802:GTP-Binding Protein alpha Subunits, Gq-G11
keywords: Allosteric Modulator; Allosteric Regulation; Arrestin; G Protein-coupled Receptor (GPCR); GPCR Regulation; Receptor Endocytosis; Trafficking
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