|Entry||Database: PDB / ID: 6ira|
|Title||Structure of the human GluN1/GluN2A NMDA receptor in the glutamate/glycine-bound state at pH 7.8|
|Keywords||MEMBRANE PROTEIN / ionotropic glutamate receptors / NMDA receptors / synaptic protein|
|Function / homology||Ionotropic glutamate receptor, metazoa / CREB1 phosphorylation through the activation of CaMKII/CaMKK/CaMKIV cascasde / N-methyl D-aspartate receptor 2B3 C-terminus / Calmodulin-binding domain C0 of NMDA receptor NR1 subunit / Ligated ion channel L-glutamate- and glycine-binding site / EPHB-mediated forward signaling / Receptor, ligand binding region / Unblocking of NMDA receptors, glutamate binding and activation / Receptor family ligand binding region / Ligand-gated ion channel ...Ionotropic glutamate receptor, metazoa / CREB1 phosphorylation through the activation of CaMKII/CaMKK/CaMKIV cascasde / N-methyl D-aspartate receptor 2B3 C-terminus / Calmodulin-binding domain C0 of NMDA receptor NR1 subunit / Ligated ion channel L-glutamate- and glycine-binding site / EPHB-mediated forward signaling / Receptor, ligand binding region / Unblocking of NMDA receptors, glutamate binding and activation / Receptor family ligand binding region / Ligand-gated ion channel / Periplasmic binding protein-like I / Ionotropic glutamate receptor, L-glutamate and glycine-binding domain / Glutamate [NMDA] receptor, epsilon subunit, C-terminal / Ras activation upon Ca2+ influx through NMDA receptor / Synaptic adhesion-like molecules / Ionotropic glutamate receptor / RAF/MAP kinase cascade / MECP2 regulates neuronal receptors and channels / Calmodulin-binding domain C0, NMDA receptor, NR1 subunit / Neurexins and neuroligins / response to glycine / propylene metabolic process / pons maturation / protein localization to postsynaptic membrane / directional locomotion / excitatory chemical synaptic transmission / regulation of respiratory gaseous exchange / conditioned taste aversion / olfactory learning / male mating behavior / serotonin metabolic process / glutamate receptor signaling pathway / startle response / regulation of synapse assembly / sleep / glutamate-gated calcium ion channel activity / neurotransmitter binding / calcium ion transmembrane import into cytosol / cation transport / neurogenesis / respiratory gaseous exchange / calcium ion homeostasis / activation of cysteine-type endopeptidase activity / glycine binding / NMDA glutamate receptor activity / glutamate binding / NMDA selective glutamate receptor complex / regulation of dendrite morphogenesis / dopamine metabolic process / suckling behavior / synaptic membrane / positive regulation of cysteine-type endopeptidase activity / social behavior / positive regulation of reactive oxygen species biosynthetic process / regulation of axonogenesis / synaptic transmission, glutamatergic / postsynaptic density membrane / positive regulation of calcium ion transport into cytosol / response to morphine / excitatory synapse / long-term memory / excitatory postsynaptic potential / synaptic cleft / positive regulation of excitatory postsynaptic potential / sensory perception of pain / prepulse inhibition / adult locomotory behavior / regulation of membrane potential / cerebral cortex development / integral component of postsynaptic density membrane / regulation of sensory perception of pain / transmitter-gated ion channel activity involved in regulation of postsynaptic membrane potential / response to amphetamine / regulation of synaptic plasticity / visual learning / long-term synaptic potentiation / ionotropic glutamate receptor signaling pathway / memory / regulation of long-term neuronal synaptic plasticity / positive regulation of long-term synaptic potentiation / presynaptic membrane / terminal bouton / synaptic vesicle / ephrin receptor signaling pathway / negative regulation of protein catabolic process / protein heterotetramerization / brain development / postsynaptic membrane / response to wounding / learning or memory / Ras guanyl-nucleotide exchange factor activity / chemical synaptic transmission / amyloid-beta binding / negative regulation of neuron apoptotic process / calmodulin binding / response to ethanol / dendritic spine / MAPK cascade / postsynaptic density / protein-containing complex binding|
Function and homology information
|Specimen source||Homo sapiens (human)|
|Method||ELECTRON MICROSCOPY / single particle reconstruction / cryo EM / 4.5 Å resolution|
|Authors||Zhang, J. / Chang, S. / Zhang, X. / Zhu, S.|
|Citation||Journal: Cell Rep / Year: 2018|
Title: Structural Basis of the Proton Sensitivity of Human GluN1-GluN2A NMDA Receptors.
Authors: Jin-Bao Zhang / Shenghai Chang / Pan Xu / Miao Miao / Hangjun Wu / Youyi Zhang / Tongtong Zhang / Han Wang / Jilin Zhang / Chun Xie / Nan Song / Cheng Luo / Xing Zhang / Shujia Zhu
Abstract: N-methyl-D-aspartate (NMDA) receptors are critical for synaptic development and plasticity. While glutamate is the primary agonist, protons can modulate NMDA receptor activity at synapses during ...N-methyl-D-aspartate (NMDA) receptors are critical for synaptic development and plasticity. While glutamate is the primary agonist, protons can modulate NMDA receptor activity at synapses during vesicle exocytosis by mechanisms that are unknown. We used cryo-electron microscopy to solve the structures of the human GluN1-GluN2A NMDA receptor at pH 7.8 and pH 6.3. Our structures demonstrate that the proton sensor predominantly resides in the N-terminal domain (NTD) of the GluN2A subunit and reveal the allosteric coupling mechanism between the proton sensor and the channel gate. Under high-pH conditions, the GluN2A-NTD adopts an "open-and-twisted" conformation. However, upon protonation at the lower pH, the GluN2A-NTD transits from an open- to closed-cleft conformation, causing rearrangements between the tetrameric NTDs and agonist-binding domains. The conformational mobility observed in our structures (presumably from protonation) is supported by molecular dynamics simulation. Our findings reveal the structural mechanisms by which protons allosterically inhibit human GluN1-GluN2A receptor activity.
SummaryFull reportAbout validation report
|Date||Deposition: Nov 12, 2018 / Release: Jan 16, 2019|
|Structure viewer||Molecule: |
Downloads & links
D: Glutamate receptor ionotropic, NMDA 2A
A: Glutamate receptor ionotropic, NMDA 1
B: Glutamate receptor ionotropic, NMDA 2A
C: Glutamate receptor ionotropic, NMDA 1
Mass: 95603.875 Da / Num. of mol.: 2 / Details: 2 mM L-Glutamate / Mutation: E656R, E657R / Source: (gene. exp.) Homo sapiens (human) / Gene: GRIN2A, NMDAR2A / Cell line (production host): HEK293S GnTI- / Production host: Homo sapiens (human) / References: UniProt: Q12879
Mass: 95336.219 Da / Num. of mol.: 2 / Details: 2 mM Glycine / Mutation: G612R / Source: (gene. exp.) Homo sapiens (human) / Gene: GRIN1, NMDAR1 / Cell line (production host): HEK293S GnTI- / Production host: Homo sapiens (human) / References: UniProt: Q05586
|Experiment||Method: ELECTRON MICROSCOPY|
|EM experiment||Aggregation state: PARTICLE / Reconstruction method: single particle reconstruction|
|Component||Name: Human GluN1/GluN2A NMDA receptors in the glutamate/glycine-bound state at pH 7.8|
Type: COMPLEX / Details: with the presence of Glycine, L-glutamate and EDTA / Entity ID: 1,
|Molecular weight||Value: 0.38 MDa / Experimental value: NO||Source (natural)||Organism: Homo sapiens (human)||Source (recombinant)||Cell: HEK293S GnTI - / Organism: Homo sapiens (human) / Plasmid: pEG-Bacmam||Buffer solution||Details: Solutions were made fresh. / pH: 7.8||Buffer component|
|Specimen||Conc.: 3.5 mg/ml / Details: Tetrameric GluN1/GluN2A NMDA receptors / Embedding applied: NO / Shadowing applied: NO / Staining applied: NO / Vitrification applied: YES||Specimen support||Details: 15 mA / Grid material: GOLD / Grid mesh size: 200 / Grid type: Quantifoil R1.2/1.3||Vitrification||Instrument: FEI VITROBOT MARK II / Cryogen name: ETHANE / Humidity: 100 % / Chamber temperature: 291 kelvins|
Details: blot for 2 seconds before plunging in liquid ethane.
-Electron microscopy imaging
Model: Titan Krios / Image courtesy: FEI Company
|Microscopy||Microscope model: FEI TITAN KRIOS|
|Electron gun||Electron source: FIELD EMISSION GUN / Accelerating voltage: 300 kV / Illumination mode: FLOOD BEAM|
|Electron lens||Mode: BRIGHT FIELDBright-field microscopy|
|Image recording||Average exposure time: 8 sec. / Electron dose: 63 e/Å2 / Detector mode: SUPER-RESOLUTION / Film or detector model: GATAN K2 SUMMIT (4k x 4k) / Number of grids imaged: 4 / Number of real images: 3171|
|Image scans||Movie frames/image: 32 / Used frames/image: 1-32|
|CTF correction||Type: PHASE FLIPPING AND AMPLITUDE CORRECTION|
|Particle selection||Number of particles selected: 431818|
|Symmetry||Point symmetry: C2|
|3D reconstruction||Resolution: 4.5 Å / Resolution method: FSC 0.143 CUT-OFF / Number of particles: 131990 / Symmetry type: POINT|
|Atomic model building||Ref protocol: RIGID BODY FIT|
|Atomic model building|
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