title: Covalent agonists for studying G protein-coupled receptor activation.
abstract: Structural studies on G protein-coupled receptors (GPCRs) provide important insights into the architecture and function of these important drug targets. However, the crystallization of GPCRs in active states is particularly challenging, requiring the formation of stable and conformationally homogeneous ligand-receptor complexes. Native hormones, neurotransmitters, and synthetic agonists that bind with low affinity are ineffective at stabilizing an active state for crystallogenesis. To promote structural studies on the pharmacologically highly relevant class of aminergic GPCRs, we here present the development of covalently binding molecular tools activating Gs-, Gi-, and Gq-coupled receptors. The covalent agonists are derived from the monoamine neurotransmitters noradrenaline, dopamine, serotonin, and histamine, and they were accessed using a general and versatile synthetic strategy. We demonstrate that the tool compounds presented herein display an efficient covalent binding mode and that the respective covalent ligand-receptor complexes activate G proteins comparable to the natural neurotransmitters. A crystal structure of the β2-adrenoreceptor in complex with a covalent noradrenaline analog and a conformationally selective antibody (nanobody) verified that these agonists can be used to facilitate crystallogenesis.
authors.affiliation: Department of Chemistry and Pharmacy, Friedrich Alexander University, 91052 Erlangen, Germany; and.
authors.forename: Dietmar
authors.initials: D
authors.lastname: Weichert
authors.affiliation: Department of Molecular and Cellular Physiology, Stanford University School of Medicine, Stanford, CA 94305.
authors.forename: Andrew C
authors.initials: AC
authors.lastname: Kruse
authors.affiliation: Department of Molecular and Cellular Physiology, Stanford University School of Medicine, Stanford, CA 94305.
authors.forename: Aashish
authors.initials: A
authors.lastname: Manglik
authors.affiliation: Department of Chemistry and Pharmacy, Friedrich Alexander University, 91052 Erlangen, Germany; and.
authors.forename: Christine
authors.initials: C
authors.lastname: Hiller
authors.affiliation: Department of Molecular and Cellular Physiology, Stanford University School of Medicine, Stanford, CA 94305.
authors.forename: Cheng
authors.initials: C
authors.lastname: Zhang
authors.affiliation: Department of Chemistry and Pharmacy, Friedrich Alexander University, 91052 Erlangen, Germany; and.
authors.forename: Harald
authors.initials: H
authors.lastname: Hübner
authors.affiliation: Department of Molecular and Cellular Physiology, Stanford University School of Medicine, Stanford, CA 94305 kobilka@stanford.edu peter.gmeiner@fau.de.
authors.forename: Brian K
authors.initials: BK
authors.lastname: Kobilka
authors.affiliation: Department of Chemistry and Pharmacy, Friedrich Alexander University, 91052 Erlangen, Germany; and kobilka@stanford.edu peter.gmeiner@fau.de.
authors.forename: Peter
authors.initials: P
authors.lastname: Gmeiner
mesh_terms: D058666:Adrenergic beta-2 Receptor Agonists / Q000737:chemistry / Q000494:pharmacology*; D006801:Humans; D008024:Ligands; D008958:Models, Molecular; D018377:Neurotransmitter Agents / Q000378:metabolism; D009638:Norepinephrine / Q000031:analogs & derivatives / Q000378:metabolism; D011869:Radioligand Assay; D018343:Receptors, Adrenergic, beta-2 / Q000737:chemistry / Q000378:metabolism*
chemical_list: D058666:Adrenergic beta-2 Receptor Agonists; D008024:Ligands; D018377:Neurotransmitter Agents; D018343:Receptors, Adrenergic, beta-2; D009638:Norepinephrine
keywords: chemical biology; chemical probes; structural biology