[English] 日本語
Yorodumi- EMDB-5203: Subtomogram average of epsilon15 using Zernike phase contrast mic... -
+Open data
-Basic information
Entry | Database: EMDB / ID: EMD-5203 | |||||||||
---|---|---|---|---|---|---|---|---|---|---|
Title | Subtomogram average of epsilon15 using Zernike phase contrast microscopy | |||||||||
Map data | This is an image of a surface rendered top-view of epsilon 15 phage reconstructed with 50 subtomograms taken with a Zernike phase plate. | |||||||||
Sample |
| |||||||||
Keywords | Phase contrast / cryoEM / electron tomography / bacteriophage | |||||||||
Biological species | epsilon15 (virus) | |||||||||
Method | subtomogram averaging / cryo EM / Resolution: 32.0 Å | |||||||||
Authors | Murata K / Liu X / Danev R / Jakana J / Schmid MF / King J / Nagayama K / Chiu W | |||||||||
Citation | Journal: Structure / Year: 2010 Title: Zernike phase contrast cryo-electron microscopy and tomography for structure determination at nanometer and subnanometer resolutions. Authors: Kazuyoshi Murata / Xiangan Liu / Radostin Danev / Joanita Jakana / Michael F Schmid / Jonathan King / Kuniaki Nagayama / Wah Chiu / Abstract: Zernike phase contrast cryo-electron microscopy (ZPC-cryoEM) is an emerging technique that is capable of producing higher image contrast than conventional cryoEM. By combining this technique with ...Zernike phase contrast cryo-electron microscopy (ZPC-cryoEM) is an emerging technique that is capable of producing higher image contrast than conventional cryoEM. By combining this technique with advanced image processing methods, we achieved subnanometer resolution for two biological specimens: 2D bacteriorhodopsin crystal and epsilon15 bacteriophage. For an asymmetric reconstruction of epsilon15 bacteriophage, ZPC-cryoEM can reduce the required amount of data by a factor of approximately 3, compared with conventional cryoEM. The reconstruction was carried out to 13 A resolution without the need to correct the contrast transfer function. New structural features at the portal vertex of the epsilon15 bacteriophage are revealed in this reconstruction. Using ZPC cryo-electron tomography (ZPC-cryoET), a similar level of data reduction and higher resolution structures of epsilon15 bacteriophage can be obtained relative to conventional cryoET. These results show quantitatively the benefits of ZPC-cryoEM and ZPC-cryoET for structural determinations of macromolecular machines at nanometer and subnanometer resolutions. | |||||||||
History |
|
-Structure visualization
Movie |
Movie viewer |
---|---|
Structure viewer | EM map: SurfViewMolmilJmol/JSmol |
Supplemental images |
-Downloads & links
-EMDB archive
Map data | emd_5203.map.gz | 36.7 MB | EMDB map data format | |
---|---|---|---|---|
Header (meta data) | emd-5203-v30.xml emd-5203.xml | 8.6 KB 8.6 KB | Display Display | EMDB header |
Images | emd_5203_1.png | 105.8 KB | ||
Archive directory | http://ftp.pdbj.org/pub/emdb/structures/EMD-5203 ftp://ftp.pdbj.org/pub/emdb/structures/EMD-5203 | HTTPS FTP |
-Validation report
Summary document | emd_5203_validation.pdf.gz | 78 KB | Display | EMDB validaton report |
---|---|---|---|---|
Full document | emd_5203_full_validation.pdf.gz | 77.1 KB | Display | |
Data in XML | emd_5203_validation.xml.gz | 493 B | Display | |
Arichive directory | https://ftp.pdbj.org/pub/emdb/validation_reports/EMD-5203 ftp://ftp.pdbj.org/pub/emdb/validation_reports/EMD-5203 | HTTPS FTP |
-Related structure data
-Links
EMDB pages | EMDB (EBI/PDBe) / EMDataResource |
---|
-Map
File | Download / File: emd_5203.map.gz / Format: CCP4 / Size: 37.5 MB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES) | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
Annotation | This is an image of a surface rendered top-view of epsilon 15 phage reconstructed with 50 subtomograms taken with a Zernike phase plate. | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Voxel size | X=Y=Z: 4.88 Å | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Density |
| ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Symmetry | Space group: 1 | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Details | EMDB XML:
CCP4 map header:
|
-Supplemental data
-Sample components
-Entire : Epsilon 15
Entire | Name: Epsilon 15 |
---|---|
Components |
|
-Supramolecule #1000: Epsilon 15
Supramolecule | Name: Epsilon 15 / type: sample / ID: 1000 / Details: The sample was monodisperse / Number unique components: 1 |
---|
-Supramolecule #1: epsilon15
Supramolecule | Name: epsilon15 / type: virus / ID: 1 / Name.synonym: epsilon15 / Sci species name: epsilon15 / Database: NCBI / Virus type: VIRION / Virus isolate: STRAIN / Virus enveloped: No / Virus empty: No / Syn species name: epsilon15 |
---|---|
Host (natural) | Organism: Salmonella (bacteria) / synonym: BACTERIA(EUBACTERIA) |
Virus shell | Shell ID: 1 / Diameter: 700 Å / T number (triangulation number): 7 |
-Experimental details
-Structure determination
Method | cryo EM |
---|---|
Processing | subtomogram averaging |
Aggregation state | particle |
-Sample preparation
Buffer | pH: 7.5 |
---|---|
Vitrification | Cryogen name: ETHANE / Instrument: OTHER |
-Electron microscopy
Microscope | JEOL 2200FS |
---|---|
Temperature | Average: 90 K |
Specialist optics | Energy filter - Name: JEOL / Energy filter - Lower energy threshold: 0.0 eV / Energy filter - Upper energy threshold: 20.0 eV |
Image recording | Number real images: 71 / Bits/pixel: 16 |
Electron beam | Acceleration voltage: 200 kV / Electron source: FIELD EMISSION GUN |
Electron optics | Illumination mode: FLOOD BEAM / Imaging mode: BRIGHT FIELD / Cs: 4.3 mm / Nominal magnification: 20000 |
Sample stage | Specimen holder: Eucentric / Specimen holder model: GATAN LIQUID NITROGEN / Tilt series - Axis1 - Min angle: -70 ° / Tilt series - Axis1 - Max angle: 70 ° |
-Image processing
Details | Average number of projections used in the 3D reconstructions: 50. |
---|---|
Final reconstruction | Resolution.type: BY AUTHOR / Resolution: 32.0 Å / Resolution method: FSC 0.5 CUT-OFF |