title: Bioorthogonal Labeling of Ghrelin Receptor to Facilitate Studies of Ligand-Dependent Conformational Dynamics.
abstract: Ghrelin receptor (GhrR) is a promising drug target because of its central role in energy homeostasis. GhrR, known for high constitutive activity, is thought to display multi-state conformations during activation and signaling. We used genetically encoded unnatural amino acids and bioorthogonal labeling reactions to engineer multiple fluorescent donor-acceptor pairs to probe ligand-directed structural changes in GhrR. We demonstrate how conformational dynamics of a G-protein-coupled receptor can be measured in reconstituted systems.
authors.affiliation: Laboratory of Chemical Biology and Signal Transduction, The Rockefeller University, 1230 York Avenue, New York, NY 10065, USA.
authors.forename: Minyoung
authors.initials: M
authors.lastname: Park
authors.affiliation: Laboratory for Molecular Pharmacology, Department of Neuroscience and Pharmacology, University of Copenhagen, The Panum Institute, Blegdamsvej 3b, 2200 Copenhagen, Denmark.
authors.forename: Bjørn B
authors.initials: BB
authors.lastname: Sivertsen
authors.affiliation: Institute of Biochemistry, Faculty of Biosciences, Pharmacy, and Psychology, Universität Leipzig, Brüderstraße 34, 04103 Leipzig, Germany.
authors.forename: Sylvia
authors.initials: S
authors.lastname: Els-Heindl
authors.affiliation: Laboratory of Chemical Biology and Signal Transduction, The Rockefeller University, 1230 York Avenue, New York, NY 10065, USA.
authors.forename: Thomas
authors.initials: T
authors.lastname: Huber
authors.affiliation: Laboratory for Molecular Pharmacology, Department of Neuroscience and Pharmacology, University of Copenhagen, The Panum Institute, Blegdamsvej 3b, 2200 Copenhagen, Denmark.
authors.forename: Birgitte
authors.initials: B
authors.lastname: Holst
authors.affiliation: Institute of Biochemistry, Faculty of Biosciences, Pharmacy, and Psychology, Universität Leipzig, Brüderstraße 34, 04103 Leipzig, Germany.
authors.forename: Annette G
authors.initials: AG
authors.lastname: Beck-Sickinger
authors.affiliation: Laboratory for Molecular Pharmacology, Department of Neuroscience and Pharmacology, University of Copenhagen, The Panum Institute, Blegdamsvej 3b, 2200 Copenhagen, Denmark.
authors.forename: Thue W
authors.initials: TW
authors.lastname: Schwartz
authors.affiliation: Laboratory of Chemical Biology and Signal Transduction, The Rockefeller University, 1230 York Avenue, New York, NY 10065, USA; Department of Neurobiology, Care Sciences and Society, Division for Neurogeriatrics, Center for Alzheimer Research, Karolinska Institutet, 141 57 Huddinge, Sweden. Electronic address: sakmar@rockefeller.edu.
authors.forename: Thomas P
authors.initials: TP
authors.lastname: Sakmar
mesh_terms: D000911:Antibodies, Monoclonal / Q000737:chemistry / Q000276:immunology; D001386:Azides / Q000737:chemistry; D061565:Cycloaddition Reaction; D031541:Fluorescence Resonance Energy Transfer; D005456:Fluorescent Dyes / Q000737:chemistry; D006801:Humans; D008024:Ligands; D009942:Organometallic Compounds / Q000737:chemistry; D010649:Phenylalanine / Q000031:analogs & derivatives / Q000737:chemistry; D011487:Protein Conformation; D054440:Receptors, Ghrelin / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism
chemical_list: D000911:Antibodies, Monoclonal; D001386:Azides; D005456:Fluorescent Dyes; D008024:Ligands; D009942:Organometallic Compounds; D054440:Receptors, Ghrelin; C070734:europium(III) trisbipyridine cryptate; C027162:4-azidophenylalanine; D010649:Phenylalanine