|Entry||Database: EMDB / ID: 8545|
|Title||SpaO mutant structure of Salmonella Type III secretion injectisomeType three secretion system|
|Map data||SpaO mutant structure of Salmonella Type III secretion injectisomeType three secretion system|
|Sample||SpaO mutant of Salmonella injectisome:|
|Method||subtomogram averaging / cryo EM / 30 Å resolution|
|Authors||Hu B / Liu J|
|Citation||Journal: Cell / Year: 2017|
Title: In Situ Molecular Architecture of the Salmonella Type III Secretion Machine.
Authors: Bo Hu / Maria Lara-Tejero / Qingke Kong / Jorge E Galán / Jun Liu
Abstract: Type III protein secretion systems have specifically evolved to deliver bacterially encoded proteins into target eukaryotic cells. The core elements of this multi-protein machine are the ...Type III protein secretion systems have specifically evolved to deliver bacterially encoded proteins into target eukaryotic cells. The core elements of this multi-protein machine are the envelope-associated needle complex, the inner membrane export apparatus, and a large cytoplasmic sorting platform. Here, we report a high-resolution in situ structure of the Salmonella Typhimurium type III secretion machine obtained by high-throughput cryo-electron tomography and sub-tomogram averaging. Through molecular modeling and comparative analysis of machines assembled with protein-tagged components or from different deletion mutants, we determined the molecular architecture of the secretion machine in situ and localized its structural components. We also show that docking of the sorting platform results in significant conformational changes in the needle complex to provide the symmetry adaptation required for the assembly of the entire secretion machine. These studies provide major insight into the structure and assembly of a broadly distributed protein secretion machine.
|Date||Deposition: Jan 4, 2017 / Header (metadata) release: Feb 8, 2017 / Map release: Mar 29, 2017 / Last update: Feb 14, 2018|
|Structure viewer||EM map: |
Downloads & links
|File||emd_8545.map.gz (map file in CCP4 format, 32001 KB)|
|Projections & slices|
Images are generated by Spider.
|Voxel size||X=Y=Z: 5.2 Å|
CCP4 map header:
-Entire SpaO mutant of Salmonella injectisome
|Entire||Name: SpaO mutant of Salmonella injectisome / Number of components: 1|
-Component #1: protein, SpaO mutant of Salmonella injectisome
|Protein||Name: SpaO mutant of Salmonella injectisome / Recombinant expression: No|
|Source||Species: Salmonella (bacteria)|
|Specimen||Specimen state: cell / Method: cryo EM|
|Sample solution||pH: 7|
|Vitrification||Cryogen name: ETHANE|
-Electron microscopy imaging
Model: Tecnai Polara / Image courtesy: FEI Company
|Imaging||Microscope: FEI POLARA 300|
|Electron gun||Electron source: FIELD EMISSION GUN / Accelerating voltage: 300 kV / Electron dose: 1.4 e/Å2 / Illumination mode: OTHER|
|Lens||Imaging mode: DARK FIELD|
|Specimen Holder||Model: OTHER / Tilt Angle: -51 - 51 deg.|
|Camera||Detector: GATAN K2 (4k x 4k)|
|Processing||Method: subtomogram averaging / Applied symmetry: C1 (asymmetric)|
|3D reconstruction||Resolution: 30 Å / Resolution method: FSC 0.5 CUT-OFF|
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