title: Mapping physiological G protein-coupled receptor signaling pathways reveals a role for receptor phosphorylation in airway contraction.
abstract: G protein-coupled receptors (GPCRs) are known to initiate a plethora of signaling pathways in vitro. However, it is unclear which of these pathways are engaged to mediate physiological responses. Here, we examine the distinct roles of Gq/11-dependent signaling and receptor phosphorylation-dependent signaling in bronchial airway contraction and lung function regulated through the M3-muscarinic acetylcholine receptor (M3-mAChR). By using a genetically engineered mouse expressing a G protein-biased M3-mAChR mutant, we reveal the first evidence, to our knowledge, of a role for M3-mAChR phosphorylation in bronchial smooth muscle contraction in health and in a disease state with relevance to human asthma. Furthermore, this mouse model can be used to distinguish the physiological responses that are regulated by M3-mAChR phosphorylation (which include control of lung function) from those responses that are downstream of G protein signaling. In this way, we present an approach by which to predict the physiological/therapeutic outcome of M3-mAChR-biased ligands with important implications for drug discovery.
authors.affiliation: Medical Research Council Toxicology Unit, University of Leicester, Leicester LE1 9HN, United Kingdom; Institute of Molecular Cell and Systems Biology, University of Glasgow, Glasgow G12 8QQ, United Kingdom;
authors.forename: Sophie J
authors.initials: SJ
authors.lastname: Bradley
authors.affiliation: Airway Disease Section, National Heart & Lung Institute, Imperial College London, London SW3 6LY, United Kingdom;
authors.forename: Coen H
authors.identifier: http://orcid.org/0000-0001-9161-0581
authors.initials: CH
authors.lastname: Wiegman
authors.affiliation: Department of Infection, Immunity, and Inflammation, Maurice Shock Medical Sciences Building, University of Leicester, Leicester LE1 9HN, United Kingdom;
authors.forename: Max Maza
authors.initials: MM
authors.lastname: Iglesias
authors.affiliation: School of Health Sciences, University of Brighton, Brighton BN2 4GL, United Kingdom;
authors.forename: Kok Choi
authors.initials: KC
authors.lastname: Kong
authors.affiliation: Medical Research Council Toxicology Unit, University of Leicester, Leicester LE1 9HN, United Kingdom; Institute of Molecular Cell and Systems Biology, University of Glasgow, Glasgow G12 8QQ, United Kingdom;
authors.forename: Adrian J
authors.initials: AJ
authors.lastname: Butcher
authors.affiliation: Institute for Research in Immunology and Cancer, Department of Biochemistry and Molecular Medicine, Université de Montréal, Montreal, QC, Canada H3C 3J7;
authors.forename: Bianca
authors.initials: B
authors.lastname: Plouffe
authors.affiliation: Department of Medicine, McGill University, Montreal, QC, Canada H3A 2B2;
authors.forename: Eugénie
authors.initials: E
authors.lastname: Goupil
authors.affiliation: Medical Research Council Toxicology Unit, University of Leicester, Leicester LE1 9HN, United Kingdom;
authors.forename: Julie-Myrtille
authors.initials: JM
authors.lastname: Bourgognon
authors.affiliation: Medical Research Council Toxicology Unit, University of Leicester, Leicester LE1 9HN, United Kingdom;
authors.forename: Timothy
authors.initials: T
authors.lastname: Macedo-Hatch
authors.affiliation: Institute for Research in Immunology and Cancer, Department of Biochemistry and Molecular Medicine, Université de Montréal, Montreal, QC, Canada H3C 3J7;
authors.forename: Christian
authors.initials: C
authors.lastname: LeGouill
authors.affiliation: Airway Disease Section, National Heart & Lung Institute, Imperial College London, London SW3 6LY, United Kingdom;
authors.forename: Kirsty
authors.initials: K
authors.lastname: Russell
authors.affiliation: Department of Medicine, McGill University, Montreal, QC, Canada H3A 2B2;
authors.forename: Stéphane A
authors.initials: SA
authors.lastname: Laporte
authors.affiliation: Department of Pharmacy, University of Bonn, 53115 Bonn, Germany.
authors.forename: Gabriele M
authors.initials: GM
authors.lastname: König
authors.affiliation: Department of Pharmacy, University of Bonn, 53115 Bonn, Germany.
authors.forename: Evi
authors.initials: E
authors.lastname: Kostenis
authors.affiliation: Institute for Research in Immunology and Cancer, Department of Biochemistry and Molecular Medicine, Université de Montréal, Montreal, QC, Canada H3C 3J7;
authors.forename: Michel
authors.initials: M
authors.lastname: Bouvier
authors.affiliation: Airway Disease Section, National Heart & Lung Institute, Imperial College London, London SW3 6LY, United Kingdom;
authors.forename: Kian Fan
authors.initials: KF
authors.lastname: Chung
authors.affiliation: Department of Infection, Immunity, and Inflammation, Maurice Shock Medical Sciences Building, University of Leicester, Leicester LE1 9HN, United Kingdom; ya26@le.ac.uk tba@le.ac.uk.
authors.forename: Yassine
authors.initials: Y
authors.lastname: Amrani
authors.affiliation: Medical Research Council Toxicology Unit, University of Leicester, Leicester LE1 9HN, United Kingdom; Institute of Molecular Cell and Systems Biology, University of Glasgow, Glasgow G12 8QQ, United Kingdom; ya26@le.ac.uk tba@le.ac.uk.
authors.forename: Andrew B
authors.initials: AB
authors.lastname: Tobin
mesh_terms: D000818:Animals; D001980:Bronchi / Q000166:cytology / Q000378:metabolism*; D006801:Humans; D051379:Mice; D018345:Mice, Knockout; D009130:Muscle, Smooth / Q000166:cytology / Q000378:metabolism*; D010766:Phosphorylation / Q000502:physiology; D043587:Receptor, Muscarinic M3 / Q000235:genetics / Q000378:metabolism*; D015398:Signal Transduction / Q000502:physiology*
chemical_list: D043587:Receptor, Muscarinic M3
keywords: G protein-coupled receptor; asthma; ligand bias; muscarinic; signaling