title: A Novel Allosteric Activator of Free Fatty Acid 2 Receptor Displays Unique Gi-functional Bias.
abstract: The short chain fatty acid receptor FFA2 is able to stimulate signaling via both Gi- and Gq/G11-promoted pathways. These pathways are believed to control distinct physiological end points but FFA2 receptor ligands appropriate to test this hypothesis have been lacking. Herein, we characterize AZ1729, a novel FFA2 regulator that acts as a direct allosteric agonist and as a positive allosteric modulator, increasing the activity of the endogenously produced short chain fatty acid propionate in Gi-mediated pathways, but not at those transduced by Gq/G11 Using AZ1729 in combination with direct inhibitors of Gi and Gq/G11 family G proteins demonstrated that although both arms contribute to propionate-mediated regulation of phospho-ERK1/2 MAP kinase signaling in FFA2-expressing 293 cells, the Gq/G11-mediated pathway is predominant. We extend these studies by employing AZ1729 to dissect physiological FFA2 signaling pathways. The capacity of AZ1729 to act at FFA2 receptors to inhibit β-adrenoreceptor agonist-promoted lipolysis in primary mouse adipocytes and to promote chemotaxis of isolated human neutrophils confirmed these as FFA2 processes mediated by Gi signaling, whereas, in concert with blockade by the Gq/G11 inhibitor FR900359, the inability of AZ1729 to mimic or regulate propionate-mediated release of GLP-1 from mouse colonic preparations defined this physiological response as an end point transduced via activation of Gq/G11.
authors.affiliation: From the Centre for Translational Pharmacology, Institute of Molecular, Cell and Systems Biology, College of Medical, Veterinary and Life Sciences, University of Glasgow, Glasgow G12 8QQ, Scotland, United Kingdom.
authors.forename: Daniele
authors.initials: D
authors.lastname: Bolognini
authors.affiliation: From the Centre for Translational Pharmacology, Institute of Molecular, Cell and Systems Biology, College of Medical, Veterinary and Life Sciences, University of Glasgow, Glasgow G12 8QQ, Scotland, United Kingdom, the Medical Research Council Toxicology Unit, University of Leicester, Leicester LE1 9HN, United Kingdom.
authors.forename: Daryl
authors.initials: D
authors.lastname: Hodge
authors.affiliation: the Cardiovascular and Metabolic Diseases.
authors.forename: Karolina
authors.initials: K
authors.lastname: Nilsson
authors.affiliation: Respiratory, Inflammatory and Autoimmune Diseases Innovative Medicines and Early Development Biotech Unit, Department of Medicinal Chemistry, and.
authors.forename: Annika U
authors.initials: AU
authors.lastname: Petersson
authors.affiliation: the Institute of Cardiovascular and Medical Sciences, College of Medical, Veterinary and Life Sciences, University of Glasgow, Glasgow G12 8QQ, Scotland, United Kingdom.
authors.forename: Iona
authors.initials: I
authors.lastname: Donnelly
authors.affiliation: From the Centre for Translational Pharmacology, Institute of Molecular, Cell and Systems Biology, College of Medical, Veterinary and Life Sciences, University of Glasgow, Glasgow G12 8QQ, Scotland, United Kingdom.
authors.forename: Eugenia
authors.initials: E
authors.lastname: Sergeev
authors.affiliation: Institute of Pharmaceutical Biology, University of Bonn, 53115 Bonn, Germany, and.
authors.forename: Gabriele M
authors.initials: GM
authors.lastname: König
authors.affiliation: the Molecular, Cellular and Pharmacobiology Section.
authors.forename: Evi
authors.initials: E
authors.lastname: Kostenis
authors.affiliation: the Institute of Infection, Immunity and Inflammation, College of Medical, Veterinary and Life Sciences, University of Glasgow, Glasgow G12 8QQ, Scotland, United Kingdom.
authors.forename: Mariola
authors.initials: M
authors.lastname: Kurowska-Stolarska
authors.affiliation: the Institute of Cardiovascular and Medical Sciences, College of Medical, Veterinary and Life Sciences, University of Glasgow, Glasgow G12 8QQ, Scotland, United Kingdom.
authors.forename: Ashley
authors.initials: A
authors.lastname: Miller
authors.affiliation: the Discovery Sciences, Reagents and Assay Development, AstraZeneca, Mölndal, Pepparedsleden 1, SE-431 83, Mölndal, Sweden.
authors.forename: Niek
authors.initials: N
authors.lastname: Dekker
authors.affiliation: From the Centre for Translational Pharmacology, Institute of Molecular, Cell and Systems Biology, College of Medical, Veterinary and Life Sciences, University of Glasgow, Glasgow G12 8QQ, Scotland, United Kingdom, the Medical Research Council Toxicology Unit, University of Leicester, Leicester LE1 9HN, United Kingdom.
authors.forename: Andrew B
authors.initials: AB
authors.lastname: Tobin
authors.affiliation: From the Centre for Translational Pharmacology, Institute of Molecular, Cell and Systems Biology, College of Medical, Veterinary and Life Sciences, University of Glasgow, Glasgow G12 8QQ, Scotland, United Kingdom, Graeme.Milligan@glasgow.ac.uk.
authors.forename: Graeme
authors.identifier: 0000-0002-6946-3519
authors.initials: G
authors.lastname: Milligan
mesh_terms: D000494:Allosteric Regulation / Q000187:drug effects; D000818:Animals; D003106:Colon / Q000378:metabolism; D047630:Depsipeptides / Q000494:pharmacology*; D019206:GTP-Binding Protein alpha Subunits, Gi-Go / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism; D043802:GTP-Binding Protein alpha Subunits, Gq-G11 / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism; D006801:Humans; D008066:Lipolysis / Q000187:drug effects / Q000235:genetics; D020935:MAP Kinase Signaling System / Q000187:drug effects* / Q000235:genetics; D051379:Mice; D018345:Mice, Knockout; D019950:Mitogen-Activated Protein Kinase 1 / Q000235:genetics / Q000378:metabolism; D048052:Mitogen-Activated Protein Kinase 3 / Q000235:genetics / Q000378:metabolism; D009504:Neutrophils / Q000378:metabolism; D011956:Receptors, Cell Surface / Q000819:agonists / Q000235:genetics / Q000378:metabolism; D043562:Receptors, G-Protein-Coupled / Q000819:agonists / Q000235:genetics / Q000378:metabolism
chemical_list: D047630:Depsipeptides; D019206:GTP-Binding Protein alpha Subunits, Gi-Go; D043802:GTP-Binding Protein alpha Subunits, Gq-G11; D019950:Mitogen-Activated Protein Kinase 1; D048052:Mitogen-Activated Protein Kinase 3; D011956:Receptors, Cell Surface; D043562:Receptors, G-Protein-Coupled; C473103:FFAR2 protein, human; C000607068:FR900359; C506481:Ffar2 protein, mouse; C554061:GNAQ protein, human; C535150:MAPK1 protein, human
keywords: G protein; G protein-coupled receptor (GPCR); allosteric regulation; fatty acid; inflammation