|Entry||Database: EMDB / ID: EMD-20713|
|Title||In vivo structure of the Legionella type II secretion system by electron cryotomography (aligning the OM-complex).Type II secretion system|
|Sample||Subtomogram average of the Legionella pneumophila type II secretion system (aligning the OM-complex).Type II secretion system|
|Biological species||Legionella pneumophila (bacteria)|
|Method||subtomogram averaging / cryo EM / Resolution: 40 Å|
|Authors||Ghosal D / Jensen GJ|
|Funding support|| United States, 1 items |
|Citation||Journal: Nat Microbiol / Year: 2019|
Title: In vivo structure of the Legionella type II secretion system by electron cryotomography.
Authors: Debnath Ghosal / Ki Woo Kim / Huaixin Zheng / Mohammed Kaplan / Hilary K Truchan / Alberto E Lopez / Ian E McIntire / Joseph P Vogel / Nicholas P Cianciotto / Grant J Jensen /
Abstract: The type II secretion system (T2SS) is a multiprotein envelope-spanning assembly that translocates a wide range of virulence factors, enzymes and effectors through the outer membrane of many Gram- ...The type II secretion system (T2SS) is a multiprotein envelope-spanning assembly that translocates a wide range of virulence factors, enzymes and effectors through the outer membrane of many Gram-negative bacteria. Here, using electron cryotomography and subtomogram averaging methods, we reveal the in vivo structure of an intact T2SS imaged within the human pathogen Legionella pneumophila. Although the T2SS has only limited sequence and component homology with the evolutionarily related type IV pilus (T4P) system, we show that their overall architectures are remarkably similar. Despite similarities, there are also differences, including, for example, that the T2SS-ATPase complex is usually present but disengaged from the inner membrane, the T2SS has a much longer periplasmic vestibule and it has a short-lived flexible pseudopilus. Placing atomic models of the components into our electron cryotomography map produced a complete architectural model of the intact T2SS that provides insights into the structure and function of its components, its position within the cell envelope and the interactions between its different subcomplexes.
|Structure viewer||EM map: |
Downloads & links
|File||Download / File: emd_20713.map.gz / Format: CCP4 / Size: 2.9 MB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES)|
|Projections & slices|
Images are generated by Spider.
generated in cubic-lattice coordinate
|Voxel size||X=Y=Z: 8.4 Å|
|Symmetry||Space group: 1|
CCP4 map header:
-Entire Subtomogram average of the Legionella pneumophila type II secreti...
|Entire||Name: Subtomogram average of the Legionella pneumophila type II secretion system (aligning the OM-complex).Type II secretion system|
Details: We used electron cryotomography and subtomogram averaging to visualize L. pneumophila type II secretion system complex directly in intact, frozen-hydrated bacterial cells
Number of components: 1
-Component #1: protein, Subtomogram average of the Legionella pneumophila type I...
|Protein||Name: Subtomogram average of the Legionella pneumophila type II secretion system (aligning the OM-complex).Type II secretion system|
Details: We used electron cryotomography and subtomogram averaging to visualize L. pneumophila type II secretion system complex directly in intact, frozen-hydrated bacterial cellsType II secretion system
Recombinant expression: No
|Source||Species: Legionella pneumophila (bacteria) / Strain: Lp02|
|Specimen||Specimen state: Cell / Method: cryo EM|
|Sample solution||pH: 6.9|
|Vitrification||Cryogen name: OTHER|
-Electron microscopy imaging
Model: Titan Krios / Image courtesy: FEI Company
|Imaging||Microscope: FEI TITAN KRIOS|
|Electron gun||Electron source: FIELD EMISSION GUN / Accelerating voltage: 300 kV / Electron dose: 1.2 e/Å2 / Illumination mode: FLOOD BEAM|
|Lens||Imaging mode: BRIGHT FIELD|
|Specimen Holder||Model: OTHER|
|Camera||Detector: GATAN K2 SUMMIT (4k x 4k)|
|Processing||Method: subtomogram averaging / Applied symmetry: C2 (2 fold cyclic) / Number of subtomograms: 440|
|3D reconstruction||Software: IMOD / Resolution: 40 Å / Resolution method: FSC 0.143 CUT-OFF|
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