Journal: Sci Rep / Year: 2019 Title: Structural and Functional Insights into GluK3-kainate Receptor Desensitization and Recovery. Authors: Jyoti Kumari / Rajesh Vinnakota / Janesh Kumar / Abstract: GluK3-kainate receptors are atypical members of the iGluR family that reside at both the pre- and postsynapse and play a vital role in the regulation of synaptic transmission. For a better ...GluK3-kainate receptors are atypical members of the iGluR family that reside at both the pre- and postsynapse and play a vital role in the regulation of synaptic transmission. For a better understanding of structural changes that underlie receptor functions, GluK3 receptors were trapped in desensitized and resting/closed states and structures analyzed using single particle cryo-electron microscopy. While the desensitized GluK3 has domain organization as seen earlier for another kainate receptor-GluK2, antagonist bound GluK3 trapped a resting state with only two LBD domains in dimeric arrangement necessary for receptor activation. Using structures as a guide, we show that the N-linked glycans at the interface of GluK3 ATD and LBD likely mediate inter-domain interactions and attune receptor-gating properties. The mutational analysis also identified putative N-glycan interacting residues. Our results provide a molecular framework for understanding gating properties unique to GluK3 and exploring the role of N-linked glycosylation in their modulation.
History
Deposition
Feb 13, 2019
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Header (metadata) release
Jul 24, 2019
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Map release
Jul 24, 2019
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Update
Nov 6, 2019
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Current status
Nov 6, 2019
Processing site: PDBj / Status: Released
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Structure visualization
Movie
Surface view with section colored by density value
Purified and detergent solubilized GluK3 receptors
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Electron microscopy
Microscope
FEI TITAN KRIOS
Image recording
Film or detector model: FEI FALCON III (4k x 4k) / Detector mode: COUNTING / Number grids imaged: 1 / Number real images: 719 / Average exposure time: 60.0 sec. / Average electron dose: 16.73 e/Å2
Electron beam
Acceleration voltage: 300 kV / Electron source: FIELD EMISSION GUN
Electron optics
Illumination mode: SPOT SCAN / Imaging mode: BRIGHT FIELD / Cs: 2.0 mm
Number classes used: 1 / Applied symmetry - Point group: C1 (asymmetric) / Algorithm: FOURIER SPACE / Resolution.type: BY AUTHOR / Resolution: 7.4 Å / Resolution method: FSC 0.143 CUT-OFF / Software - Name: cryoSPARC (ver. 1) / Software - details: final reconstruction / Number images used: 9730
Initial angle assignment
Type: COMMON LINE / Software - Name: cryoSPARC (ver. 1) / Software - details: Abinitio 3D reconstruction / Details: Abinitio 3D reconstruction module of cryosparc
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