title: Glutamate receptor desensitization is mediated by changes in quaternary structure of the ligand binding domain.
abstract: Glutamate receptor ion channels are membrane proteins that mediate excitatory synaptic transmission in the central nervous system of vertebrates. Insight into molecular mechanisms underlying glutamate receptor gating is limited by lack of structural information for receptors trapped in different conformational states. Here, we report the use of single-particle cryoelectron tomography to determine the structures, at ∼21 Å resolution, of full-length GluK2 kainate receptors trapped in antagonist-bound resting and agonist-bound desensitized states. The resting state, stabilized by the competitive antagonist LY466195, closely resembles the crystal structure of the AMPA receptor GluA2, with well-resolved proximal and distal subunits exhibiting cross-over between the twofold symmetric amino terminal domain and a twofold symmetric ligand binding domain (LBD) dimer of dimers assembly. In the desensitized state, the LBD undergoes a major rearrangement, resulting in a separation of the four subunits by ∼25 Å. However, the amino terminal domain, transmembrane, and cytoplasmic regions of the receptor have similar conformations in the resting and desensitized states. The LBD rearrangement was not anticipated in prior models based on crystal structures for soluble LBD dimer assemblies, and we speculate that subunit separation allows a better match to the fourfold symmetric ion channel domain. From fits of the amino terminal domain and LBD domains into the density map of the desensitized state we have derived a structural model for differences in quaternary conformation between the resting and desensitized states.
authors.affiliation: Laboratory of Cell Biology, Center for Cancer Research, National Cancer Institute, National Institutes of Health, Bethesda, MD 20892, USA.
authors.forename: David M
authors.initials: DM
authors.lastname: Schauder
authors.forename: Oleg
authors.initials: O
authors.lastname: Kuybeda
authors.forename: Jinjin
authors.initials: J
authors.lastname: Zhang
authors.forename: Katherine
authors.initials: K
authors.lastname: Klymko
authors.forename: Alberto
authors.initials: A
authors.lastname: Bartesaghi
authors.forename: Mario J
authors.initials: MJ
authors.lastname: Borgnia
authors.forename: Mark L
authors.initials: ML
authors.lastname: Mayer
authors.forename: Sriram
authors.initials: S
authors.lastname: Subramaniam
mesh_terms: D000818:Animals; D001665:Binding Sites; D020285:Cryoelectron Microscopy; D018360:Crystallography, X-Ray; D015640:Ion Channel Gating; D008024:Ligands; D008958:Models, Molecular; D017354:Point Mutation; D054730:Protein Interaction Domains and Motifs; D020836:Protein Structure, Quaternary; D051381:Rats; D018092:Kainic Acid Receptors / Q000737:chemistry*; D014054:Tomography; D000097806:GluK2 Kainate Receptor
chemical_list: D008024:Ligands; D018092:Kainic Acid Receptors; D000097806:GluK2 Kainate Receptor