title: A G Protein-biased Designer G Protein-coupled Receptor Useful for Studying the Physiological Relevance of Gq/11-dependent Signaling Pathways.
abstract: Designerreceptorsexclusivelyactivated by adesignerdrug (DREADDs) are clozapine-N-oxide-sensitive designer G protein-coupled receptors (GPCRs) that have emerged as powerful novel chemogenetic tools to study the physiological relevance of GPCR signaling pathways in specific cell types or tissues. Like endogenous GPCRs, clozapine-N-oxide-activated DREADDs do not only activate heterotrimeric G proteins but can also trigger β-arrestin-dependent (G protein-independent) signaling. To dissect the relative physiological relevance of G protein-mediatedversusβ-arrestin-mediated signaling in different cell types or physiological processes, the availability of G protein- and β-arrestin-biased DREADDs would be highly desirable. In this study, we report the development of a mutationally modified version of a non-biased DREADD derived from the M3muscarinic receptor that can activate Gq/11with high efficacy but lacks the ability to interact with β-arrestins. We also demonstrate that this novel DREADD is activein vivoand that cell type-selective expression of this new designer receptor can provide novel insights into the physiological roles of G protein (Gq/11)-dependentversusβ-arrestin-dependent signaling in hepatocytes. Thus, this novel Gq/11-biased DREADD represents a powerful new tool to study the physiological relevance of Gq/11-dependent signaling in distinct tissues and cell types, in the absence of β-arrestin-mediated cellular effects. Such studies should guide the development of novel classes of functionally biased ligands that show high efficacy in various pathophysiological conditions but display a reduced incidence of side effects.
authors: From the Molecular Signaling Section, Laboratory of Bioorganic Chemistry, NIDDK, National Institutes of Health, Bethesda, Maryland 20892.
authors: Jianxin
authors: J
authors: Hu
authors: From the Molecular Signaling Section, Laboratory of Bioorganic Chemistry, NIDDK, National Institutes of Health, Bethesda, Maryland 20892.
authors: Matthew
authors: M
authors: Stern
authors: the Vanderbilt University Medical Center, Nashville, Tennessee 37232.
authors: Luis E
authors: LE
authors: Gimenez
authors: the Institute of Biochemistry, University of Leipzig, Leipzig 04103, Germany.
authors: Lizzy
authors: L
authors: Wanka
authors: From the Molecular Signaling Section, Laboratory of Bioorganic Chemistry, NIDDK, National Institutes of Health, Bethesda, Maryland 20892.
authors: Lu
authors: L
authors: Zhu
authors: From the Molecular Signaling Section, Laboratory of Bioorganic Chemistry, NIDDK, National Institutes of Health, Bethesda, Maryland 20892.
authors: Mario
authors: M
authors: Rossi
authors: From the Molecular Signaling Section, Laboratory of Bioorganic Chemistry, NIDDK, National Institutes of Health, Bethesda, Maryland 20892.
authors: Jaroslawna
authors: J
authors: Meister
authors: the Graduate School of Pharmaceutical Science, Tohoku University, Sendai, Miyagi 980-8578, Japan, and the Japan Science and Technology Agency, Precursory Research for Embryonic Science and Technology (PRESTO), Kawaguchi, Saitama 332-0012, Japan.
authors: Asuka
authors: A
authors: Inoue
authors: the Institute of Biochemistry, University of Leipzig, Leipzig 04103, Germany.
authors: Annette G
authors: AG
authors: Beck-Sickinger
authors: the Vanderbilt University Medical Center, Nashville, Tennessee 37232.
authors: Vsevolod V
authors: VV
authors: Gurevich
authors: From the Molecular Signaling Section, Laboratory of Bioorganic Chemistry, NIDDK, National Institutes of Health, Bethesda, Maryland 20892, jwess@helix.nih.gov.
authors: Jürgen
authors: J
authors: Wess
mesh_terms: D000818:Animals; D019390:Arrestins / Q000378:metabolism; D019556:COS Cells; D002118:Calcium / Q000378:metabolism; D002478:Cells, Cultured; D002522:Chlorocebus aethiops; D043802:GTP-Binding Protein alpha Subunits, Gq-G11 / Q000235:genetics / Q000378:metabolism*; D055785:Gene Knockdown Techniques; D005947:Glucose / Q000378:metabolism; D057809:HEK293 Cells; D022781:Hepatocytes / Q000378:metabolism*; D006801:Humans; D008810:Mice, Inbred C57BL; D025941:Protein Interaction Mapping / Q000379:methods; D060066:Protein Interaction Maps; D043562:Receptors, G-Protein-Coupled / Q000378:metabolism*; D015398:Signal Transduction; D000071557:beta-Arrestins
chemical_list: D019390:Arrestins; D043562:Receptors, G-Protein-Coupled; D000071557:beta-Arrestins; D043802:GTP-Binding Protein alpha Subunits, Gq-G11; D005947:Glucose; D002118:Calcium
keywords: 7-helix receptor; DREADD; G protein; G protein-coupled receptor (GPCR); arrestin; biased signaling; cell signaling