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Yorodumi- PDB-8qb9: Helical reconstruction of yeast eisosome protein Pil1 bound to me... -
+Open data
-Basic information
Entry | Database: PDB / ID: 8qb9 | |||||||||||||||
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Title | Helical reconstruction of yeast eisosome protein Pil1 bound to membrane composed of lipid mixture -PIP2/+sterol (DOPC, DOPE, DOPS, cholesterol 30:20:20:30) | |||||||||||||||
Components | Sphingolipid long chain base-responsive protein PIL1 | |||||||||||||||
Keywords | LIPID BINDING PROTEIN / BAR domain / lipid reconstitution / membrane microdomain | |||||||||||||||
Function / homology | Function and homology information protein localization to eisosome filament / eisosome filament / eisosome assembly / eisosome / lipid droplet / cell periphery / protein localization / endocytosis / mitochondrial outer membrane / lipid binding ...protein localization to eisosome filament / eisosome filament / eisosome assembly / eisosome / lipid droplet / cell periphery / protein localization / endocytosis / mitochondrial outer membrane / lipid binding / mitochondrion / plasma membrane / cytoplasm Similarity search - Function | |||||||||||||||
Biological species | Saccharomyces cerevisiae (brewer's yeast) | |||||||||||||||
Method | ELECTRON MICROSCOPY / helical reconstruction / cryo EM / Resolution: 3.35 Å | |||||||||||||||
Authors | Kefauver, J.M. / Zou, L. / Desfosses, A. / Loewith, R.J. | |||||||||||||||
Funding support | European Union, Switzerland, 4items
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Citation | Journal: To Be Published Title: CryoEM architecture of a native stretch-sensitive membrane microdomain Authors: Kefauver, J.M. / Hakala, M. / Zou, L. / Alba, J. / Espadas Moreno, J. / Tettamanti, M.G. / Estrozi, L. / Vanni, S. / Roux, A. / Desfosses, A. / Loewith, R. | |||||||||||||||
History |
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-Structure visualization
Structure viewer | Molecule: MolmilJmol/JSmol |
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-Downloads & links
-Download
PDBx/mmCIF format | 8qb9.cif.gz | 831.6 KB | Display | PDBx/mmCIF format |
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PDB format | pdb8qb9.ent.gz | 559.1 KB | Display | PDB format |
PDBx/mmJSON format | 8qb9.json.gz | Tree view | PDBx/mmJSON format | |
Others | Other downloads |
-Validation report
Summary document | 8qb9_validation.pdf.gz | 1.7 MB | Display | wwPDB validaton report |
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Full document | 8qb9_full_validation.pdf.gz | 1.7 MB | Display | |
Data in XML | 8qb9_validation.xml.gz | 99 KB | Display | |
Data in CIF | 8qb9_validation.cif.gz | 131.9 KB | Display | |
Arichive directory | https://data.pdbj.org/pub/pdb/validation_reports/qb/8qb9 ftp://data.pdbj.org/pub/pdb/validation_reports/qb/8qb9 | HTTPS FTP |
-Related structure data
Related structure data | 18308MC 8qb7C 8qb8C 8qbbC 8qbdC 8qbeC 8qbfC 8qbgC 8r1lC M: map data used to model this data C: citing same article (ref.) |
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Similar structure data | Similarity search - Function & homologyF&H Search |
-Links
-Assembly
Deposited unit |
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Noncrystallographic symmetry (NCS) | NCS domain:
NCS domain segments: Component-ID: 1 / Ens-ID: ens_1 / Beg auth comp-ID: MET / Beg label comp-ID: MET / End auth comp-ID: THR / End label comp-ID: THR / Auth seq-ID: 1 - 271 / Label seq-ID: 1 - 271
NCS oper:
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-Components
#1: Protein | Mass: 38393.043 Da / Num. of mol.: 14 Source method: isolated from a genetically manipulated source Source: (gene. exp.) Saccharomyces cerevisiae (brewer's yeast) Strain: TB50 / Gene: PIL1 / Plasmid: pCoofy6 Details (production host): gift from Sabine Suppmann, Addgene plasmid #43990 Production host: Escherichia coli 'BL21-Gold(DE3)pLysS AG' (bacteria) References: UniProt: P53252 |
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-Experimental details
-Experiment
Experiment | Method: ELECTRON MICROSCOPY |
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EM experiment | Aggregation state: HELICAL ARRAY / 3D reconstruction method: helical reconstruction |
-Sample preparation
Component | Name: Helical assembly of recombinant Pil1 protein tubulating -PIP2/+sterol lipid mixture (DOPC,DOPE,DOPS,cholesterol 30:20:20:30) Type: COMPLEX / Entity ID: all / Source: RECOMBINANT |
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Molecular weight | Experimental value: NO |
Source (natural) | Organism: Saccharomyces cerevisiae (brewer's yeast) / Strain: TB50 |
Source (recombinant) | Organism: Escherichia coli 'BL21-Gold(DE3)pLysS AG' (bacteria) Plasmid: pCoofy6 |
Buffer solution | pH: 7.4 / Details: 20mM HEPES, pH 7.4, 150mM KoAc, 2mM MgAc |
Specimen | Embedding applied: NO / Shadowing applied: NO / Staining applied: NO / Vitrification applied: YES |
Specimen support | Grid material: COPPER / Grid mesh size: 300 divisions/in. / Grid type: EMS Lacey Carbon |
Vitrification | Instrument: LEICA EM GP / Cryogen name: ETHANE / Humidity: 90 % / Chamber temperature: 291 K |
-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 defocus max: 1800 nm / Nominal defocus min: 600 nm / Cs: 2.7 mm |
Image recording | Electron dose: 50 e/Å2 / Film or detector model: FEI FALCON IV (4k x 4k) |
-Processing
EM software |
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CTF correction | Type: PHASE FLIPPING AND AMPLITUDE CORRECTION | |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Helical symmerty |
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3D reconstruction | Resolution: 3.35 Å / Resolution method: FSC 0.143 CUT-OFF / Num. of particles: 176005 / Symmetry type: HELICAL | |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Atomic model building | Space: REAL | |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Atomic model building | Accession code: P53252 / Source name: AlphaFold / Type: in silico model | |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Refinement | Cross valid method: NONE Stereochemistry target values: GeoStd + Monomer Library + CDL v1.2 | |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Displacement parameters | Biso mean: 84.74 Å2 | |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Refine LS restraints |
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Refine LS restraints NCS |
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