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Yorodumi- PDB-8x0e: Human FL Metabotropic glutamate receptor 5, mGlu5-5M with agonist... -
+Open data
-Basic information
Entry | Database: PDB / ID: 8x0e | ||||||||||||
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Title | Human FL Metabotropic glutamate receptor 5, mGlu5-5M with agonist and PAM, W785A mutant | ||||||||||||
Components | Metabotropic glutamate receptor 5 | ||||||||||||
Keywords | MEMBRANE PROTEIN / G-PROTEIN COUPLED RECEPTORS / SIGNAL TRANSDUCTION / METABOTROPIC GLUTAMATE RECEPTOR / Agonist / active state | ||||||||||||
Function / homology | Function and homology information A2A adenosine receptor binding / phospholipase C-activating G protein-coupled glutamate receptor signaling pathway / G protein-coupled receptor activity involved in regulation of postsynaptic membrane potential / adenylate cyclase inhibiting G protein-coupled glutamate receptor activity / trans-synaptic signaling by endocannabinoid, modulating synaptic transmission / neurotransmitter receptor activity involved in regulation of postsynaptic cytosolic calcium ion concentration / positive regulation of long-term neuronal synaptic plasticity / desensitization of G protein-coupled receptor signaling pathway / G protein-coupled glutamate receptor signaling pathway / astrocyte projection ...A2A adenosine receptor binding / phospholipase C-activating G protein-coupled glutamate receptor signaling pathway / G protein-coupled receptor activity involved in regulation of postsynaptic membrane potential / adenylate cyclase inhibiting G protein-coupled glutamate receptor activity / trans-synaptic signaling by endocannabinoid, modulating synaptic transmission / neurotransmitter receptor activity involved in regulation of postsynaptic cytosolic calcium ion concentration / positive regulation of long-term neuronal synaptic plasticity / desensitization of G protein-coupled receptor signaling pathway / G protein-coupled glutamate receptor signaling pathway / astrocyte projection / Class C/3 (Metabotropic glutamate/pheromone receptors) / : / glutamate receptor activity / Neurexins and neuroligins / protein tyrosine kinase activator activity / regulation of synaptic transmission, glutamatergic / positive regulation of calcium-mediated signaling / protein tyrosine kinase binding / learning / dendritic shaft / locomotory behavior / G protein-coupled receptor activity / postsynaptic density membrane / synapse organization / Schaffer collateral - CA1 synapse / cognition / cellular response to amyloid-beta / chemical synaptic transmission / G alpha (q) signalling events / positive regulation of MAPK cascade / dendritic spine / learning or memory / glutamatergic synapse / dendrite / regulation of DNA-templated transcription / identical protein binding / plasma membrane / cytoplasm Similarity search - Function | ||||||||||||
Biological species | Homo sapiens (human) | ||||||||||||
Method | ELECTRON MICROSCOPY / single particle reconstruction / cryo EM / Resolution: 3.4 Å | ||||||||||||
Authors | Vinothkumar, K.R. / Lebon, G. / Cannone, G. | ||||||||||||
Funding support | India, France, 3items
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Citation | Journal: To Be Published Title: Conformational diversity in class C GPCR positive allosteric modulation Authors: Vinothkumar, K.R. / Lebon, G. / Cannone, G. | ||||||||||||
History |
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-Structure visualization
Structure viewer | Molecule: MolmilJmol/JSmol |
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-Downloads & links
-Download
PDBx/mmCIF format | 8x0e.cif.gz | 314.2 KB | Display | PDBx/mmCIF format |
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PDB format | pdb8x0e.ent.gz | 248.2 KB | Display | PDB format |
PDBx/mmJSON format | 8x0e.json.gz | Tree view | PDBx/mmJSON format | |
Others | Other downloads |
-Validation report
Summary document | 8x0e_validation.pdf.gz | 1.3 MB | Display | wwPDB validaton report |
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Full document | 8x0e_full_validation.pdf.gz | 1.2 MB | Display | |
Data in XML | 8x0e_validation.xml.gz | 59.6 KB | Display | |
Data in CIF | 8x0e_validation.cif.gz | 89.3 KB | Display | |
Arichive directory | https://data.pdbj.org/pub/pdb/validation_reports/x0/8x0e ftp://data.pdbj.org/pub/pdb/validation_reports/x0/8x0e | HTTPS FTP |
-Related structure data
Related structure data | 37976MC 8x0bC 8x0cC 8x0dC 8x0fC 8x0gC 8x0hC C: citing same article (ref.) M: map data used to model this data |
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Similar structure data | Similarity search - Function & homologyF&H Search |
-Links
-Assembly
Deposited unit |
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1 |
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-Components
#1: Protein | Mass: 93619.383 Da / Num. of mol.: 2 / Mutation: T742A,S753A,T777A,I799A,A813L,W785A,H350L Source method: isolated from a genetically manipulated source Details: The N-terminal sequence (DYKDDDDKHHHHHHHHHHLEVLFQGP) is the tag and linker), which has been cleaved by protease before structural experiment. Here, it is included for completion. The ...Details: The N-terminal sequence (DYKDDDDKHHHHHHHHHHLEVLFQGP) is the tag and linker), which has been cleaved by protease before structural experiment. Here, it is included for completion. The sequence when compared to uniprot starts at 21 and ends at 856 (construct used for expression). The construct has the following mutations (with reference to Uniprot ID P41594) H350L - mutation for nanobody binding T742A, S753A, T777A, I799A, A813L (thermostabilising mutant) W785A - introduced mutant to test for PAM binding. There are 10 disulfide bonds per chain and 2 NAG molecules. Source: (gene. exp.) Homo sapiens (human) / Gene: GRM5 / Cell line (production host): HEK293S GnT- / Production host: Homo sapiens (human) / References: UniProt: P41594 #2: Sugar | #3: Chemical | Has ligand of interest | Y | Has protein modification | Y | |
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-Experimental details
-Experiment
Experiment | Method: ELECTRON MICROSCOPY |
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EM experiment | Aggregation state: PARTICLE / 3D reconstruction method: single particle reconstruction |
-Sample preparation
Component | Name: Thermostabilised full length human mGluR5, W785A mutant Type: COMPLEX Details: Receptor was purified with orthosteric Quisqualate and PAM VU0424465 Entity ID: #1 / Source: RECOMBINANT | |||||||||||||||||||||||||
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Molecular weight | Value: 0.2 MDa / Experimental value: NO | |||||||||||||||||||||||||
Source (natural) | Organism: Homo sapiens (human) | |||||||||||||||||||||||||
Source (recombinant) | Organism: Homo sapiens (human) / Strain: HEK293S GnT- | |||||||||||||||||||||||||
Buffer solution | pH: 7.4 | |||||||||||||||||||||||||
Buffer component |
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Specimen | Conc.: 6 mg/ml / Embedding applied: NO / Shadowing applied: NO / Staining applied: NO / Vitrification applied: YES Details: Receptor is purified in detergent micelles and monodisperse. The receptor is purified with 10 uM Quisqualate and 10 uM PAM VU0424465 | |||||||||||||||||||||||||
Specimen support | Grid material: GOLD / Grid mesh size: 300 divisions/in. / Grid type: Quantifoil R1.2/1.3 | |||||||||||||||||||||||||
Vitrification | Instrument: FEI VITROBOT MARK IV / Cryogen name: ETHANE / Humidity: 100 % / Chamber temperature: 289 K / Details: Blot force was set to 0 |
-Electron microscopy imaging
Experimental equipment | Model: Titan Krios / Image courtesy: FEI Company |
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Microscopy | Model: FEI TITAN KRIOS |
Electron gun | Electron source: FIELD EMISSION GUN / Accelerating voltage: 300 kV / Illumination mode: FLOOD BEAM |
Electron lens | Mode: BRIGHT FIELD / Nominal magnification: 75000 X / Calibrated magnification: 130841 X / Nominal defocus max: 2800 nm / Nominal defocus min: 1800 nm / Cs: 2.7 mm / C2 aperture diameter: 50 µm / Alignment procedure: COMA FREE |
Specimen holder | Cryogen: NITROGEN / Specimen holder model: FEI TITAN KRIOS AUTOGRID HOLDER |
Image recording | Average exposure time: 60 sec. / Electron dose: 28.25 e/Å2 / Detector mode: COUNTING / Film or detector model: FEI FALCON III (4k x 4k) / Num. of grids imaged: 2 / Num. of real images: 5762 Details: Images were collected in movie mode for 60 seconds and 25 frames were stored. |
Image scans | Sampling size: 14 µm / Width: 4096 / Height: 4096 |
-Processing
EM software |
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CTF correction | Type: PHASE FLIPPING AND AMPLITUDE CORRECTION | |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Particle selection | Num. of particles selected: 783666 Details: The particles were extract from two batches of data collection | |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Symmetry | Point symmetry: C1 (asymmetric) | |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
3D reconstruction | Resolution: 3.4 Å / Resolution method: FSC 0.143 CUT-OFF / Num. of particles: 188960 / Algorithm: BACK PROJECTION / Num. of class averages: 3 / Symmetry type: POINT | |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Atomic model building | B value: 183.8 / Protocol: OTHER / Space: RECIPROCAL | |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Atomic model building | Accession code: D_1300042377 Details: The initial model was from this deposition ID, which has been submitted Source name: Other / Type: experimental model | |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Refinement | Resolution: 3.4→177.62 Å / Cor.coef. Fo:Fc: 0.977 / SU B: 11.347 / SU ML: 0.18 / ESU R: 0.261 Stereochemistry target values: MAXIMUM LIKELIHOOD WITH PHASES Details: HYDROGENS HAVE BEEN USED IF PRESENT IN THE INPUT
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Solvent computation | Solvent model: PARAMETERS FOR MASK CACLULATION | |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Displacement parameters | Biso mean: 182.715 Å2 | |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Refinement step | Cycle: 1 / Total: 12086 | |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Refine LS restraints |
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