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Yorodumi- EMDB-0070: Structure of the type IV pilus from enterohemorrhagic Escherichia... -
+Open data
-Basic information
Entry | Database: EMDB / ID: EMD-0070 | |||||||||
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Title | Structure of the type IV pilus from enterohemorrhagic Escherichia coli (EHEC) | |||||||||
Map data | ||||||||||
Sample |
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Function / homology | : / Prokaryotic N-terminal methylation site. / Prokaryotic N-terminal methylation motif / Prokaryotic N-terminal methylation site / Pilin-like / membrane / Prepilin peptidase-dependent pilin Function and homology information | |||||||||
Biological species | Escherichia coli O157:H7 (bacteria) | |||||||||
Method | helical reconstruction / cryo EM / Resolution: 8.0 Å | |||||||||
Authors | Zheng W / Egelman E / Bardiaux B / Luna-Rico A / Izadi-Pruneyre N / Francetic O | |||||||||
Funding support | France, 1 items
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Citation | Journal: Structure / Year: 2019 Title: Structure and Assembly of the Enterohemorrhagic Escherichia coli Type 4 Pilus. Authors: Benjamin Bardiaux / Gisele Cardoso de Amorim / Areli Luna Rico / Weili Zheng / Ingrid Guilvout / Camille Jollivet / Michael Nilges / Edward H Egelman / Nadia Izadi-Pruneyre / Olivera Francetic / Abstract: Bacterial type 4a pili are dynamic surface filaments that promote bacterial adherence, motility, and macromolecular transport. Their genes are highly conserved among enterobacteria and their ...Bacterial type 4a pili are dynamic surface filaments that promote bacterial adherence, motility, and macromolecular transport. Their genes are highly conserved among enterobacteria and their expression in enterohemorrhagic Escherichia coli (EHEC) promotes adhesion to intestinal epithelia and pro-inflammatory signaling. To define the molecular basis of EHEC pilus assembly, we determined the structure of the periplasmic domain of its major subunit PpdD (PpdDp), a prototype of an enterobacterial pilin subfamily containing two disulfide bonds. The structure of PpdDp, determined by NMR, was then docked into the density envelope of purified EHEC pili obtained by cryoelectron microscopy (cryo-EM). Cryo-EM reconstruction of EHEC pili at ∼8 Å resolution revealed extremely high pilus flexibility correlating with a large extended region of the pilin stem. Systematic mutagenesis combined with functional and interaction analyses identified charged residues essential for pilus assembly. Structural information on exposed regions and interfaces between EHEC pilins is relevant for vaccine and drug discovery. | |||||||||
History |
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-Structure visualization
Movie |
Movie viewer |
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Structure viewer | EM map: SurfViewMolmilJmol/JSmol |
Supplemental images |
-Downloads & links
-EMDB archive
Map data | emd_0070.map.gz | 3 MB | EMDB map data format | |
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Header (meta data) | emd-0070-v30.xml emd-0070.xml | 11.6 KB 11.6 KB | Display Display | EMDB header |
Images | emd_0070.png | 35.1 KB | ||
Archive directory | http://ftp.pdbj.org/pub/emdb/structures/EMD-0070 ftp://ftp.pdbj.org/pub/emdb/structures/EMD-0070 | HTTPS FTP |
-Validation report
Summary document | emd_0070_validation.pdf.gz | 207.9 KB | Display | EMDB validaton report |
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Full document | emd_0070_full_validation.pdf.gz | 207 KB | Display | |
Data in XML | emd_0070_validation.xml.gz | 5.9 KB | Display | |
Arichive directory | https://ftp.pdbj.org/pub/emdb/validation_reports/EMD-0070 ftp://ftp.pdbj.org/pub/emdb/validation_reports/EMD-0070 | HTTPS FTP |
-Related structure data
Related structure data | 6gv9MC 6gmsC M: atomic model generated by this map C: citing same article (ref.) |
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Similar structure data |
-Links
EMDB pages | EMDB (EBI/PDBe) / EMDataResource |
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Related items in Molecule of the Month |
-Map
File | Download / File: emd_0070.map.gz / Format: CCP4 / Size: 27 MB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES) | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
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Projections & slices | Image control
Images are generated by Spider. | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Voxel size | X=Y=Z: 1.09 Å | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Density |
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Symmetry | Space group: 1 | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Details | EMDB XML:
CCP4 map header:
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-Supplemental data
-Sample components
-Entire : EHEC type IV pili
Entire | Name: EHEC type IV pili |
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Components |
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-Supramolecule #1: EHEC type IV pili
Supramolecule | Name: EHEC type IV pili / type: complex / ID: 1 / Parent: 0 / Macromolecule list: all |
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Source (natural) | Organism: Escherichia coli O157:H7 (bacteria) |
Recombinant expression | Organism: Escherichia coli BL21(DE3) (bacteria) |
-Macromolecule #1: Prepilin peptidase-dependent protein D
Macromolecule | Name: Prepilin peptidase-dependent protein D / type: protein_or_peptide / ID: 1 / Number of copies: 14 / Enantiomer: LEVO |
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Source (natural) | Organism: Escherichia coli O157:H7 (bacteria) |
Molecular weight | Theoretical: 14.883821 KDa |
Recombinant expression | Organism: Escherichia coli BL21(DE3) (bacteria) |
Sequence | String: FTLIELMVVI GIIAILSAIG IPAYQNYLRK AALTDMLQTF VPYRTAVELC ALEHGGLDTC DGGSNGIPSP TTTRYVSAMS VAKGVVSLT GQESLNGLSV VMTPGWDNAN GVTGWARNCN IQSDSALQQA CEDVFRFDDA N |
-Experimental details
-Structure determination
Method | cryo EM |
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Processing | helical reconstruction |
Aggregation state | filament |
-Sample preparation
Buffer | pH: 7.4 |
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Grid | Material: COPPER / Mesh: 300 / Support film - Material: CARBON / Support film - topology: LACEY / Pretreatment - Type: PLASMA CLEANING |
Vitrification | Cryogen name: ETHANE / Chamber humidity: 95 % / Chamber temperature: 297 K / Instrument: FEI VITROBOT MARK IV |
-Electron microscopy
Microscope | FEI TITAN KRIOS |
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Image recording | Film or detector model: FEI FALCON III (4k x 4k) / Detector mode: INTEGRATING / Number grids imaged: 1 / Average exposure time: 2.0 sec. / Average electron dose: 1.4 e/Å2 |
Electron beam | Acceleration voltage: 300 kV / Electron source: FIELD EMISSION GUN |
Electron optics | Illumination mode: FLOOD BEAM / Imaging mode: BRIGHT FIELD |
Experimental equipment | Model: Titan Krios / Image courtesy: FEI Company |
-Image processing
Final reconstruction | Applied symmetry - Helical parameters - Δz: 11.2 Å Applied symmetry - Helical parameters - Δ&Phi: 96 ° Applied symmetry - Helical parameters - Axial symmetry: C1 (asymmetric) Resolution.type: BY AUTHOR / Resolution: 8.0 Å / Resolution method: FSC 0.143 CUT-OFF / Software - Name: SPIDER / Number images used: 25669 |
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CTF correction | Software - Name: CTFFIND (ver. 3) |
Startup model | Type of model: OTHER / Details: featureless cylinder |
Final angle assignment | Type: NOT APPLICABLE |
-Atomic model buiding 1
Details | Soluble domain structure and homology model of TM segment fitted as rigid-bodies. Linker region built ab initio. Real-space refinement after symmetrization. |
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Refinement | Space: REAL / Protocol: OTHER |
Output model | PDB-6gv9: |