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Yorodumi- PDB-8twv: ELIC5 with Propylamine in spNW15 nanodiscs with 2:1:1 POPC:POPE:POPG -
+Open data
-Basic information
Entry | Database: PDB / ID: 8twv | ||||||
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Title | ELIC5 with Propylamine in spNW15 nanodiscs with 2:1:1 POPC:POPE:POPG | ||||||
Components | Gamma-aminobutyric-acid receptor subunit beta-1 | ||||||
Keywords | TRANSPORT PROTEIN / ELIC / ion channel / pLGIC / Structural Protein / Membrane Protein | ||||||
Function / homology | Function and homology information extracellular ligand-gated monoatomic ion channel activity / regulation of membrane potential / transmembrane signaling receptor activity / neuron projection / signal transduction / identical protein binding / membrane Similarity search - Function | ||||||
Biological species | Dickeya dadantii (bacteria) | ||||||
Method | ELECTRON MICROSCOPY / single particle reconstruction / cryo EM / Resolution: 3.4 Å | ||||||
Authors | Dalal, V. / Arcario, M.J. / Petroff II, J.T. / Deitzen, N.M. / Tan, B.K. / Brannigan, G. / Cheng, W.W.L. | ||||||
Funding support | United States, 1items
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Citation | Journal: Nat Commun / Year: 2024 Title: Lipid nanodisc scaffold and size alter the structure of a pentameric ligand-gated ion channel. Authors: Vikram Dalal / Mark J Arcario / John T Petroff / Brandon K Tan / Noah M Dietzen / Michael J Rau / James A J Fitzpatrick / Grace Brannigan / Wayland W L Cheng / Abstract: Lipid nanodiscs have become a standard tool for studying membrane proteins, including using single particle cryo-electron microscopy (cryo-EM). We find that reconstituting the pentameric ligand-gated ...Lipid nanodiscs have become a standard tool for studying membrane proteins, including using single particle cryo-electron microscopy (cryo-EM). We find that reconstituting the pentameric ligand-gated ion channel (pLGIC), Erwinia ligand-gated ion channel (ELIC), in different nanodiscs produces distinct structures by cryo-EM. The effect of the nanodisc on ELIC structure extends to the extracellular domain and agonist binding site. Additionally, molecular dynamic simulations indicate that nanodiscs of different size impact ELIC structure and that the nanodisc scaffold directly interacts with ELIC. These findings suggest that the nanodisc plays a crucial role in determining the structure of pLGICs, and that reconstitution of ion channels in larger nanodiscs may better approximate a lipid membrane environment. | ||||||
History |
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-Structure visualization
Structure viewer | Molecule: MolmilJmol/JSmol |
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-Downloads & links
-Download
PDBx/mmCIF format | 8twv.cif.gz | 310.7 KB | Display | PDBx/mmCIF format |
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PDB format | pdb8twv.ent.gz | 254.8 KB | Display | PDB format |
PDBx/mmJSON format | 8twv.json.gz | Tree view | PDBx/mmJSON format | |
Others | Other downloads |
-Validation report
Summary document | 8twv_validation.pdf.gz | 1.2 MB | Display | wwPDB validaton report |
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Full document | 8twv_full_validation.pdf.gz | 1.2 MB | Display | |
Data in XML | 8twv_validation.xml.gz | 54.2 KB | Display | |
Data in CIF | 8twv_validation.cif.gz | 71.8 KB | Display | |
Arichive directory | https://data.pdbj.org/pub/pdb/validation_reports/tw/8twv ftp://data.pdbj.org/pub/pdb/validation_reports/tw/8twv | HTTPS FTP |
-Related structure data
Related structure data | 41672MC 8f32C 8f33C 8f34C 8f35C 8twzC 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: 37011.055 Da / Num. of mol.: 5 Source method: isolated from a genetically manipulated source Source: (gene. exp.) Dickeya dadantii (bacteria) / Gene: Dda3937_00520 / Production host: Escherichia coli (E. coli) / References: UniProt: E0SJQ4 #2: Chemical | ChemComp-3CN / Has ligand of interest | N | |
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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: ELIC5 with Propylamine in spNW15 nanodiscs with 2:1:1 POPC:POPE:POPG Type: COMPLEX / Entity ID: #1 / Source: RECOMBINANT |
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Source (natural) | Organism: Dickeya dadantii (bacteria) |
Source (recombinant) | Organism: Escherichia coli (E. coli) |
Buffer solution | pH: 7.5 |
Specimen | Embedding applied: NO / Shadowing applied: NO / Staining applied: NO / Vitrification applied: YES |
Specimen support | Details: O2 27.5 sccm H2 6.4 sccm / Grid material: COPPER / Grid mesh size: 300 divisions/in. / Grid type: Quantifoil R2/2 |
Vitrification | Instrument: FEI VITROBOT MARK IV / Cryogen name: ETHANE / Humidity: 100 % / Chamber temperature: 277 K / Details: Blot for 2 seconds before plunging |
-Electron microscopy imaging
Microscopy | Model: TFS GLACIOS |
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Electron gun | Electron source: FIELD EMISSION GUN / Accelerating voltage: 200 kV / Illumination mode: FLOOD BEAM |
Electron lens | Mode: BRIGHT FIELD / Nominal magnification: 120000 X / Calibrated magnification: 120000 X / Nominal defocus max: 2400 nm / Nominal defocus min: 1000 nm / Cs: 2.7 mm / C2 aperture diameter: 50 µm / Alignment procedure: ZEMLIN TABLEAU |
Specimen holder | Cryogen: NITROGEN / Specimen holder model: FEI TITAN KRIOS AUTOGRID HOLDER |
Image recording | Average exposure time: 11.78 sec. / Electron dose: 50.09 e/Å2 / Film or detector model: FEI FALCON IV (4k x 4k) / Num. of grids imaged: 1 |
-Processing
CTF correction | Type: PHASE FLIPPING AND AMPLITUDE CORRECTION |
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3D reconstruction | Resolution: 3.4 Å / Resolution method: FSC 0.143 CUT-OFF / Num. of particles: 27201 / Symmetry type: POINT |
Atomic model building | Protocol: AB INITIO MODEL / Space: REAL |