|Entry||Database: EMDB / ID: EMD-0432|
|Title||Architecture and subunit arrangement of native AMPA receptors elucidated by cryo-EM|
|Sample||Native A2AxA3A2 complex bound with MPQX|
|Biological species||Rattus norvegicus (Norway rat)|
|Method||single particle reconstruction / cryo EM / Resolution: 21 Å|
|Authors||Zhao Y / Gouaux E|
|Citation||Journal: Science / Year: 2019|
Title: Architecture and subunit arrangement of native AMPA receptors elucidated by cryo-EM.
Authors: Yan Zhao / Shanshuang Chen / Adam C Swensen / Wei-Jun Qian / Eric Gouaux /
Abstract: Glutamate-gated AMPA receptors mediate the fast component of excitatory signal transduction at chemical synapses throughout all regions of the mammalian brain. AMPA receptors are tetrameric ...Glutamate-gated AMPA receptors mediate the fast component of excitatory signal transduction at chemical synapses throughout all regions of the mammalian brain. AMPA receptors are tetrameric assemblies composed of four subunits, GluA1-GluA4. Despite decades of study, the subunit composition, subunit arrangement, and molecular structure of native AMPA receptors remain unknown. Here we elucidate the structures of 10 distinct native AMPA receptor complexes by single-particle cryo-electron microscopy (cryo-EM). We find that receptor subunits are arranged nonstochastically, with the GluA2 subunit preferentially occupying the B and D positions of the tetramer and with triheteromeric assemblies comprising a major population of native AMPA receptors. Cryo-EM maps define the structure for S2-M4 linkers between the ligand-binding and transmembrane domains, suggesting how neurotransmitter binding is coupled to ion channel gating.
|Structure viewer||EM map: |
Downloads & links
|File||Download / File: emd_0432.map.gz / Format: CCP4 / Size: 125 MB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES)|
|Projections & slices|
Images are generated by Spider.
|Voxel size||X=Y=Z: 1.72 Å|
|Symmetry||Space group: 1|
CCP4 map header:
-Entire Native A2AxA3A2 complex bound with MPQX
|Entire||Name: Native A2AxA3A2 complex bound with MPQX / Number of components: 1|
-Component #1: protein, Native A2AxA3A2 complex bound with MPQX
|Protein||Name: Native A2AxA3A2 complex bound with MPQX / Recombinant expression: No|
|Source||Species: Rattus norvegicus (Norway rat)|
|Specimen||Specimen state: Particle / Method: cryo EM|
|Sample solution||Specimen conc.: 4 mg/mL / pH: 8|
|Vitrification||Cryogen name: ETHANE / Temperature: 295 K / Humidity: 100 %|
-Electron microscopy imaging
Model: Titan Krios / Image courtesy: FEI Company
|Imaging||Microscope: FEI TITAN KRIOS|
|Electron gun||Electron source: FIELD EMISSION GUN / Accelerating voltage: 300 kV / Electron dose: 54 e/Å2 / Illumination mode: FLOOD BEAM|
|Lens||Imaging mode: BRIGHT FIELD|
|Specimen Holder||Model: OTHER|
|Camera||Detector: GATAN K2 SUMMIT (4k x 4k)|
|Processing||Method: single particle reconstruction / Number of projections: 10800|
|3D reconstruction||Software: RELION / Resolution: 21 Å / Resolution method: FSC 0.143 CUT-OFF|
-Atomic model buiding
|Modeling #1||Refinement protocol: rigid body / Refinement space: REAL|
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