|Entry||Database: EMDB / ID: 3596|
|Title||The structure of the ESX-5 mycobacterial type VII secretion system membrane complex|
|Sample||ESX-5 T7SS membrane complex|
|Source||Mycobacterium xenopi rivm700367 / bacteria|
|Method||single particle reconstruction, at 13.1 Å resolution|
|Authors||Ciccarelli L / Marlovits TC|
|Citation||Nat Microbiol, 2017, 2, 17047-17047|
Nat Microbiol, 2017, 2, 17047-17047 Yorodumi Papers
|Date||Deposition: Feb 17, 2017 / Header (metadata) release: Apr 5, 2017 / Map release: Apr 12, 2017 / Last update: Aug 2, 2017|
Downloads & links
|File||emd_3596.map.gz (map file in CCP4 format, 131073 KB)|
|Projections & slices|
Images are generated by Spider package.
|Voxel size||X=Y=Z: 2.18 Å|
CCP4 map header:
-Entire ESX-5 T7SS membrane complex
|Entire||Name: ESX-5 T7SS membrane complex / Details: Solubilized in amphipol A8-35 / Number of components: 1|
|Mass||Theoretical: 1.8 MDa|
-Component #1: protein, ESX-5 T7SS membrane complex
|Protein||Name: ESX-5 T7SS membrane complex / Details: Solubilized in amphipol A8-35 / Recombinant expression: No|
|Mass||Theoretical: 1.8 MDa|
|Source||Species: Mycobacterium xenopi rivm700367 / bacteria / Strain: RIVM700367|
|Source (engineered)||Expression System: Mycobacterium smegmatis str. mc2 155 / bacteria|
|Sample solution||Specimen conc.: 0.1 mg/ml|
Buffer solution: 20 mM Tris pH 8, 100 mM NaCl, 5 % (v/v) glycerol
|Staining||A drop of 2% (w/v) uranyl acetate was added to a carbon-coated grid with absorbed protein and blotted after 30 s.|
|Vitrification||Cryogen name: NONE|
-Electron microscopy imaging
|Imaging||Microscope: FEI TECNAI 20|
|Electron gun||Electron source: LAB6 / Accelerating voltage: 200 kV / Electron dose: 20 e/Å2 / Illumination mode: FLOOD BEAM|
|Lens||Magnification: 50000 X (nominal), 44000 X (calibrated) / Cs: 1.2 mm / Imaging mode: BRIGHT FIELD / Defocus: 1400 - 5000 nm|
|Specimen Holder||Model: SIDE ENTRY, EUCENTRIC|
|Camera||Detector: FEI EAGLE (4k x 4k)|
|Image acquisition||Number of digital images: 65|
|Processing||Method: single particle reconstruction / Applied symmetry: C6 (6 fold cyclic) / Number of projections: 1418|
|3D reconstruction||Algorithm: BACK PROJECTION / Software: EMAN / Resolution: 13.1 Å / Resolution method: FSC 0.143 CUT-OFF|
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Three pioneers of this field were awarded Nobel Prize in Chemistry 2017
- Jacques Dubochet (University of Lausanne, Switzerland) is a pioneer of ice-embedding method of EM specimen (as known as cryo-EM), Most of 3DEM structures in EMDB and PDB are obtained using his method.
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External links: The 2017 Nobel Prize in Chemistry - Press Release
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