+Open data
-Basic information
Entry | Database: PDB / ID: 6x80 | |||||||||
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Title | Structure of the Campylobacter jejuni G508A Flagellar Filament | |||||||||
Components | Flagellin A | |||||||||
Keywords | STRUCTURAL PROTEIN / Helical Symmetry / Bacterial Flagellar Filament | |||||||||
Function / homology | Function and homology information bacterial-type flagellum / structural molecule activity / extracellular region Similarity search - Function | |||||||||
Biological species | Campylobacter jejuni (Campylobacter) | |||||||||
Method | ELECTRON MICROSCOPY / helical reconstruction / cryo EM / Resolution: 3.5 Å | |||||||||
Authors | Kreutzberger, M.A.B. / Wang, F. / Egelman, E.H. | |||||||||
Funding support | United States, 2items
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Citation | Journal: Proc Natl Acad Sci U S A / Year: 2020 Title: Atomic structure of the flagellar filament reveals how ε Proteobacteria escaped Toll-like receptor 5 surveillance. Authors: Mark A B Kreutzberger / Cheryl Ewing / Frederic Poly / Fengbin Wang / Edward H Egelman / Abstract: Vertebrates, from zebra fish to humans, have an innate immune recognition of many bacterial flagellins. This involves a conserved eight-amino acid epitope in flagellin recognized by the Toll-like ...Vertebrates, from zebra fish to humans, have an innate immune recognition of many bacterial flagellins. This involves a conserved eight-amino acid epitope in flagellin recognized by the Toll-like receptor 5 (TLR5). Several important human pathogens, such as and , have escaped TLR5 activation by mutations in this epitope. When such mutations were introduced into flagellin, motility was abolished. It was previously argued, using very low-resolution cryoelectron microscopy (cryo-EM), that accommodated these mutations by forming filaments with 7 protofilaments, rather than the 11 found in other bacteria. We have now determined the atomic structure of the G508A flagellar filament from a 3.5-Å-resolution cryo-EM reconstruction, and show that it has 11 protofilaments. The residues in the TLR5 epitope have reduced contacts with the adjacent subunit compared to other bacterial flagellar filament structures. The weakening of the subunit-subunit interface introduced by the mutations in the TLR5 epitope is compensated for by extensive interactions between the outer domains of the flagellin subunits. In other bacteria, these outer domains can be nearly absent or removed without affecting motility. Furthermore, we provide evidence for the stabilization of these outer domain interactions through glycosylation of key residues. These results explain the essential role of glycosylation in motility, and show how the outer domains have evolved to play a role not previously found in other bacteria. | |||||||||
History |
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-Structure visualization
Movie |
Movie viewer |
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Structure viewer | Molecule: MolmilJmol/JSmol |
-Downloads & links
-Download
PDBx/mmCIF format | 6x80.cif.gz | 2 MB | Display | PDBx/mmCIF format |
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PDB format | pdb6x80.ent.gz | 1.7 MB | Display | PDB format |
PDBx/mmJSON format | 6x80.json.gz | Tree view | PDBx/mmJSON format | |
Others | Other downloads |
-Validation report
Summary document | 6x80_validation.pdf.gz | 17.9 MB | Display | wwPDB validaton report |
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Full document | 6x80_full_validation.pdf.gz | 18.2 MB | Display | |
Data in XML | 6x80_validation.xml.gz | 359.2 KB | Display | |
Data in CIF | 6x80_validation.cif.gz | 507.1 KB | Display | |
Arichive directory | https://data.pdbj.org/pub/pdb/validation_reports/x8/6x80 ftp://data.pdbj.org/pub/pdb/validation_reports/x8/6x80 | HTTPS FTP |
-Related structure data
Related structure data | 22088MC M: map data used to model this data C: citing same article (ref.) |
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Similar structure data |
-Links
-Assembly
Deposited unit |
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1 |
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-Components
#1: Protein | Mass: 59590.762 Da / Num. of mol.: 22 / Mutation: G508A Source method: isolated from a genetically manipulated source Source: (gene. exp.) Campylobacter jejuni (Campylobacter) / Gene: flaA / Production host: Campylobacter jejuni (Campylobacter) / References: UniProt: P22251 #2: Sugar | ChemComp-P8E / Has ligand of interest | Y | |
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-Experimental details
-Experiment
Experiment | Method: ELECTRON MICROSCOPY |
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EM experiment | Aggregation state: FILAMENT / 3D reconstruction method: helical reconstruction |
-Sample preparation
Component | Name: Bacterial Flagellar Filament / Type: ORGANELLE OR CELLULAR COMPONENT / Entity ID: #1 / Source: MULTIPLE SOURCES |
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Source (natural) | Organism: Campylobacter jejuni (Campylobacter) |
Source (recombinant) | Organism: Campylobacter jejuni (Campylobacter) |
Buffer solution | pH: 7 |
Specimen | Embedding applied: NO / Shadowing applied: NO / Staining applied: NO / Vitrification applied: YES |
Vitrification | Cryogen name: ETHANE |
-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 |
Image recording | Electron dose: 51 e/Å2 / Film or detector model: GATAN K3 (6k x 4k) |
-Processing
Software | Name: PHENIX / Version: 1.15.2_3472: / Classification: refinement | ||||||||||||||||||||||||
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EM software |
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CTF correction | Type: PHASE FLIPPING AND AMPLITUDE CORRECTION | ||||||||||||||||||||||||
Helical symmerty | Angular rotation/subunit: 65.32 ° / Axial rise/subunit: 4.8 Å / Axial symmetry: C1 | ||||||||||||||||||||||||
3D reconstruction | Resolution: 3.5 Å / Resolution method: OTHER / Num. of particles: 116959 / Details: MODEL:MAP FSC using a threshold of 0.5 / Symmetry type: HELICAL | ||||||||||||||||||||||||
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