title: Ligand binding to the human MT2 melatonin receptor: the role of residues in transmembrane domains 3, 6, and 7.
abstract: To better understand the mechanism of interactions between G-protein-coupled melatonin receptors and their ligands, our previously reported homology model of human MT2 receptor with docked 2-iodomelatonin was further refined and used to select residues within TM3, TM6, and TM7 potentially important for receptor-ligand interactions. Selected residues were mutated and radioligand-binding assay was used to test the binding affinities of hMT2 receptors transiently expressed in HEK293 cells. Our data demonstrate that residues N268 and A275 in TM6 as well as residues V291 and L295 in TM7 are essential for 2-iodomelatonin binding to the hMT2 receptor, while TM3 residues M120, G121, V124, and I125 may participate in binding of other receptor agonists and/or antagonists. Presented data also hint at possible specific interaction between the side-chain of Y188 in second extracellular loop and N-acetyl group of 2-iodomelatonin.
authors.affiliation: Institute of Physiology, Academy of Sciences of the Czech Republic, Videnska 1083, 142 20 Prague, Czech Republic.
authors.forename: Petr
authors.initials: P
authors.lastname: Mazna
authors.forename: Karel
authors.initials: K
authors.lastname: Berka
authors.forename: Irena
authors.initials: I
authors.lastname: Jelinkova
authors.forename: Ales
authors.initials: A
authors.lastname: Balik
authors.forename: Petr
authors.initials: P
authors.lastname: Svoboda
authors.forename: Veronika
authors.initials: V
authors.lastname: Obsilova
authors.forename: Tomas
authors.initials: T
authors.lastname: Obsil
authors.forename: Jan
authors.initials: J
authors.lastname: Teisinger
mesh_terms: D019943:Amino Acid Substitution; D001665:Binding Sites / Q000235:genetics; D002460:Cell Line; D006801:Humans; D066298:In Vitro Techniques; D007700:Kinetics; D008024:Ligands; D008550:Melatonin / Q000031:analogs & derivatives* / Q000378:metabolism; D008958:Models, Molecular; D016297:Mutagenesis, Site-Directed; D017434:Protein Structure, Tertiary; D044123:Receptor, Melatonin, MT2 / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism
chemical_list: D008024:Ligands; D044123:Receptor, Melatonin, MT2; D011994:Recombinant Proteins; C044556:2-iodomelatonin; D008550:Melatonin