title: Structural determinants of agonist-specific kinetics at the ionotropic glutamate receptor 2.
abstract: Glutamate receptors (GluRs) are the most abundant mediators of the fast excitatory neurotransmission in the human brain. Agonists will, after activation of the receptors, induce different degrees of desensitization. The efficacy of agonists strongly correlates with the agonist-induced closure of the ligand-binding domain. However, the differences in desensitization properties are less well understood. By using high-resolution x-ray structure of the GluR2 flop (GluR2o) ligand-binding core protein in complex with the partial glutamate receptor agonist (S)-2-amino-3-(3-hydroxy-5-tert-butyl-4-isothiazolyl)propionic acid [(S)-thio-ATPA], we show that (S)-thio-ATPA induces an 18 degrees closure of the binding core similar to another partial agonist, (S)-2-amino-3-(4-bromo-3-hydroxy-5-isoxazolyl)propionic acid [(S)-Br-HIBO]. Despite the similar closure of the ligand-binding domain, we find in electrophysiological studies that (S)-thio-ATPA induced a 6.4-fold larger steady-state current than (RS)-Br-HIBO, and rapid agonist applications show that (S)-thio-ATPA induces a 3.6-fold higher steady-state/peak ratio and a 2.2-fold slower desensitization time constant than (RS)-Br-HIBO. Structural comparisons reveal that (S)-Br-HIBO, but not (S)-thio-ATPA, induces a twist of the ligand-binding core compared with the apostructure, and the agonist-specific conformation of Leu-650 correlates with the different kinetic profiles pointing at a key role in defining the desensitization kinetics. We conclude that, especially for intermediate efficacious agonists, the desensitization properties are influenced by additional ligand-induced factors beyond domain closure.
authors.affiliation: Department of Molecular Biology, C. F. Møllers Allé Building 130, University of Aarhus, DK-8000 Aarhus, Denmark.
authors.forename: Mai Marie
authors.initials: MM
authors.lastname: Holm
authors.forename: Marie-Louise
authors.initials: ML
authors.lastname: Lunn
authors.forename: Stephen F
authors.initials: SF
authors.lastname: Traynelis
authors.forename: Jette S
authors.initials: JS
authors.lastname: Kastrup
authors.forename: Jan
authors.initials: J
authors.lastname: Egebjerg
mesh_terms: D000409:Alanine / Q000031:analogs & derivatives / Q000378:metabolism; D017398:Alternative Splicing / Q000235:genetics; D000818:Animals; D001921:Brain / Q000378:metabolism*; D003460:Crystallization; D004305:Dose-Response Relationship, Drug; D007051:Ibotenic Acid / Q000031:analogs & derivatives / Q000378:metabolism; D007700:Kinetics; D008958:Models, Molecular; D016296:Mutagenesis; D009865:Oocytes; D018408:Patch-Clamp Techniques; D011485:Protein Binding; D017434:Protein Structure, Tertiary; D018091:Receptors, AMPA / Q000819:agonists / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D009435:Synaptic Transmission / Q000502:physiology*; D013844:Thiazoles / Q000378:metabolism; D014982:Xenopus laevis
chemical_list: C428244:2-amino-3-(5-tert-butyl-3-hydroxy-4-isothiazolyl)propionic acid; D018091:Receptors, AMPA; D013844:Thiazoles; C023366:4-bromohomoibotenic acid; D007051:Ibotenic Acid; D000409:Alanine; C104722:glutamate receptor ionotropic, AMPA 2