|Entry||Database: EMDB / ID: 4185|
|Title||Human GroEL obtained by grid-blotting from blue native PAGE|
|Map data||Grid blotted from blue native PAGE human GroEL|
|Source||Homo sapiens (human)|
|Method||single particle reconstruction / negative staining / 25 Å resolution|
|Authors||Kirykowicz AM / Woodward JD|
|Citation||Journal: To Be Published|
Title: Human GroEL obtained by grid-blotting from blue native PAGE
Authors: Kirykowicz AM / Woodward JD
|Date||Deposition: Nov 30, 2017 / Header (metadata) release: Dec 13, 2017 / Map release: Dec 5, 2018 / Last update: Dec 5, 2018|
|Structure viewer||EM map: |
Downloads & links
|File||emd_4185.map.gz (map file in CCP4 format, 1049 KB)|
|Projections & slices|
Images are generated by Spider.
|Voxel size||X=Y=Z: 4.22 Å|
CCP4 map header:
-Entire GroEL only
|Entire||Name: GroEL only / Details: Obtained from Sigma-Aldrich / Number of components: 1|
-Component #1: protein, GroEL only
|Protein||Name: GroEL only / Details: Obtained from Sigma-Aldrich / Recombinant expression: No|
|Mass||Experimental: 840 kDa|
|Source||Species: Homo sapiens (human)|
|Source (engineered)||Expression System: Escherichia coli (E. coli)|
|Specimen||Specimen state: particle / Method: negative staining|
|Sample solution||Specimen conc.: 1 mg/ml|
Buffer solution: Diluted in 50 mM Tris-HCl, 200 mM NaCl (pH 8.0). 1-5 ug of GroEL run on blue native page: Anode buffer: 25 mM Imidazole Cathode buffer A: 50 mM Tricine, 7.5 mM Imidazole, 0.02% Coomassie G250 (pH 7.0) Cathode buffer B: 50 mM Tricine, 7.5 mM Imidazole, 0.002% Coomassie G250 (pH 7.0)
|Staining||1-5 ug of groEL was run on a blue native PAGE gel. Sample was transferred to grid by grid blotting procedure and washed/stained with 2% UA.|
|Vitrification||Cryogen name: NONE|
-Electron microscopy imaging
Model: Tecnai F20 / Image courtesy: FEI Company
|Imaging||Microscope: FEI TECNAI F20|
|Electron gun||Electron source: FIELD EMISSION GUN / Accelerating voltage: 200 kV / Electron dose: 2 e/Å2 / Illumination mode: OTHER|
|Lens||Magnification: 50000.0 X (nominal) / Cs: 1.2 mm / Imaging mode: BRIGHT FIELD|
|Specimen Holder||Model: OTHER|
|Camera||Detector: GATAN ULTRASCAN 4000 (4k x 4k)|
|Processing||Method: single particle reconstruction / Applied symmetry: D7 (2*7 fold dihedral) / Number of projections: 463|
|3D reconstruction||Resolution: 25 Å / Resolution method: FSC 0.5 CUT-OFF / Details: EMAN procedure|
Euler angles: EMAN procedure: 5 iteration of 10 degrees (19 classes generated), 5 iterations of 8 degrees (23 classes generated), 10 iterations of 5 degrees (61 classes generated), and 6 iterations of 3 degrees (155 classes generated)
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