+Open data
-Basic information
Entry | Database: PDB / ID: 8eht | ||||||
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Title | Cryo-EM reconstruction of the CS20 bacterial adhesion pili | ||||||
Components | CS20 fimbria major subunit protein | ||||||
Keywords | CELL ADHESION / enterotoxigenic / adhesion pili / superelastic | ||||||
Function / homology | Fimbrial-type adhesion domain superfamily / Adhesion domain superfamily / pilus / cell adhesion / CS20 fimbria major subunit protein Function and homology information | ||||||
Biological species | Escherichia coli (E. coli) | ||||||
Method | ELECTRON MICROSCOPY / helical reconstruction / cryo EM / Resolution: 3.4 Å | ||||||
Authors | Doran, M.H. / Bullitt, E. | ||||||
Funding support | United States, 1items
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Citation | Journal: Structure / Year: 2023 Title: Three structural solutions for bacterial adhesion pilus stability and superelasticity. Authors: Matthew H Doran / Joseph L Baker / Tobias Dahlberg / Magnus Andersson / Esther Bullitt / Abstract: Bacterial adhesion pili are key virulence factors that mediate host-pathogen interactions in diverse epithelial environments. Deploying a multimodal approach, we probed the structural basis ...Bacterial adhesion pili are key virulence factors that mediate host-pathogen interactions in diverse epithelial environments. Deploying a multimodal approach, we probed the structural basis underpinning the biophysical properties of pili originating from enterotoxigenic (ETEC) and uropathogenic bacteria. Using cryo-electron microscopy we solved the structures of three vaccine target pili from ETEC bacteria, CFA/I, CS17, and CS20. Pairing these and previous pilus structures with force spectroscopy and steered molecular dynamics simulations, we find a strong correlation between subunit-subunit interaction energies and the force required for pilus unwinding, irrespective of genetic similarity. Pili integrate three structural solutions for stabilizing their assemblies: layer-to-layer interactions, N-terminal interactions to distant subunits, and extended loop interactions from adjacent subunits. Tuning of these structural solutions alters the biophysical properties of pili and promotes the superelastic behavior that is essential for sustained bacterial attachment. | ||||||
History |
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-Structure visualization
Structure viewer | Molecule: MolmilJmol/JSmol |
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-Downloads & links
-Download
PDBx/mmCIF format | 8eht.cif.gz | 185.4 KB | Display | PDBx/mmCIF format |
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PDB format | pdb8eht.ent.gz | 153.6 KB | Display | PDB format |
PDBx/mmJSON format | 8eht.json.gz | Tree view | PDBx/mmJSON format | |
Others | Other downloads |
-Validation report
Summary document | 8eht_validation.pdf.gz | 1.2 MB | Display | wwPDB validaton report |
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Full document | 8eht_full_validation.pdf.gz | 1.2 MB | Display | |
Data in XML | 8eht_validation.xml.gz | 40.4 KB | Display | |
Data in CIF | 8eht_validation.cif.gz | 62.4 KB | Display | |
Arichive directory | https://data.pdbj.org/pub/pdb/validation_reports/eh/8eht ftp://data.pdbj.org/pub/pdb/validation_reports/eh/8eht | HTTPS FTP |
-Related structure data
Related structure data | 28152MC 8ehrC 8ehsC 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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1 |
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-Components
#1: Protein | Mass: 17435.199 Da / Num. of mol.: 7 Source method: isolated from a genetically manipulated source Source: (gene. exp.) Escherichia coli (E. coli) / Gene: csnA / Production host: Escherichia coli (E. coli) / Strain (production host): WS7179A / References: UniProt: Q8VL73 |
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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: CS20 bacterial adhesion pili / Type: ORGANELLE OR CELLULAR COMPONENT / Entity ID: all / Source: RECOMBINANT |
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Source (natural) | Organism: Escherichia coli (E. coli) / Strain: WS7179A |
Source (recombinant) | Organism: Escherichia coli (E. coli) / Strain: WS7179A-2(pRA101) |
Buffer solution | pH: 7.4 |
Specimen | Embedding applied: NO / Shadowing applied: NO / Staining applied: NO / Vitrification applied: YES |
Specimen support | Details: 15 mA using the Pelco EasiGlow machine / Grid material: GOLD / Grid mesh size: 400 divisions/in. / Grid type: UltrAuFoil R1.2/1.3 |
Vitrification | Instrument: FEI VITROBOT MARK III / Cryogen name: ETHANE / Humidity: 100 % / Chamber temperature: 283 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: 3000 nm / Nominal defocus min: 1000 nm |
Image recording | Electron dose: 53.62 e/Å2 / Film or detector model: GATAN K3 (6k x 4k) / Num. of real images: 5236 |
-Processing
EM software |
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CTF correction | Type: PHASE FLIPPING AND AMPLITUDE CORRECTION | ||||||||||||||||||||||||||||||||||||||||||||
Helical symmerty | Angular rotation/subunit: 111.84 ° / Axial rise/subunit: 8.953 Å / Axial symmetry: C1 | ||||||||||||||||||||||||||||||||||||||||||||
Particle selection | Num. of particles selected: 9590890 | ||||||||||||||||||||||||||||||||||||||||||||
3D reconstruction | Resolution: 3.4 Å / Resolution method: FSC 0.143 CUT-OFF / Num. of particles: 88048 / Symmetry type: HELICAL |