|Entry||Database: EMDB / ID: 5288|
|Title||The single particle reconstruction of the human Toll-like receptor 5 ectodomain in detergent|
|Keywords||Toll-like receptor 5 / ectodomain|
|Sample||toll-like receptor 5 ectodomain|
|Source||Homo sapiens / human|
|Map data||This is the volume of the hTLR5 ectodomain.|
|Method||single particle reconstruction, at 26 Å resolution|
|Authors||Zhou K / Kanai R / Lee P / Wang HW / Modis Y|
|Citation||J. Struct. Biol., 2012, 177, 402-409|
|Date||Deposition: May 2, 2011 / Header (metadata) release: May 5, 2011 / Map release: Dec 19, 2011 / Last update: Dec 19, 2011|
Downloads & links
|File||emd_5288.map.gz (map file in CCP4 format, 3908 KB)|
|Projections & slices|
Images are generated by Spider package.
|Voxel size||X=Y=Z: 2.14 Å|
CCP4 map header:
-Entire toll-like receptor 5 ectodomain
|Entire||Name: toll-like receptor 5 ectodomain / Details: The sample was monodisperse / Number of components: 2 / Oligomeric State: dimer|
|Mass||Theoretical: 200 kDa / Experimental: 200 kDa / Measured by: gel filtration|
-Component #1: protein, Toll-like receptor
|Protein||Name: Toll-like receptor / a.k.a: TLR5 / Oligomeric Details: dimer / Recombinant expression: Yes / Number of Copies: 2|
|Mass||Theoretical: 200 kDa / Experimental: 200 kDa|
|Source||Species: Homo sapiens / human|
|Source (engineered)||Expression System: Sf9 insect cells / Vector: pAcGP67-A|
|Source (natural)||Organelle: membrane / Location in cell: plasma membrane / Cell: Sf9 insect cells|
|External references||InterPro: InterPro: 017241|
|Sample solution||Specimen conc.: 0.005 mg/ml / Buffer solution: 10 mM TEA, 0.15 M NaCl / pH: 7.5|
|Support film||thin carbon covered home-made holey carbon on 400 mesh copper grid|
|Staining||The protein solution was applied onto a thin-carbon-covered holey carbon copper grid and negatively stained in 1% uranyl-formate solution for 1 minute.|
|Vitrification||Instrument: NONE / Cryogen name: NONE|
-Electron microscopy imaging
|Imaging||Microscope: FEI TECNAI 12 / Date: Aug 1, 2010|
|Electron gun||Electron source: LAB6 / Accelerating voltage: 120 kV / Illumination mode: FLOOD BEAM|
|Lens||Magnification: 52000 X (nominal), 52000 X (calibrated) / Cs: 2.2 mm / Imaging mode: BRIGHT FIELD / Defocus: 800 - 1200 nm|
|Specimen Holder||Holder: Eucentric / Model: SIDE ENTRY, EUCENTRIC|
|Camera||Detector: GATAN ULTRASCAN 4000 (4k x 4k)|
|Image acquisition||Number of digital images: 50|
|Processing||Method: single particle reconstruction / Number of class averages: 100 / Number of projections: 3087 / Applied symmetry: C1 (asymmetric)|
|3D reconstruction||Algorithm: Random conical tilt followed by projection matching refinement|
Software: IMAGIC SPIDER
Details: Final maps were calculated from all the particles by back-projection reconstruction.
Resolution: 26 Å / Resolution method: FSC 0.5
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