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Yorodumi- EMDB-9269: Single-Molecule 3D Image of Low-Density Lipoprotein in Complex wi... -
+Open data
-Basic information
Entry | Database: EMDB / ID: EMD-9269 | |||||||||
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Title | Single-Molecule 3D Image of Low-Density Lipoprotein in Complex with Cholesteryl Ester Transfer Protein (No. 02) | |||||||||
Map data | ||||||||||
Sample |
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Biological species | Homo sapiens (human) | |||||||||
Method | electron tomography / cryo EM / Resolution: 71.0 Å | |||||||||
Authors | Wu H / Zhai X / Lei D / Liu J / Yu Y / Bie R / Ren G | |||||||||
Citation | Journal: Sci Rep / Year: 2018 Title: An Algorithm for Enhancing the Image Contrast of Electron Tomography. Authors: Hao Wu / Xiaobo Zhai / Dongsheng Lei / Jianfang Liu / Yadong Yu / Rongfang Bie / Gang Ren / Abstract: Three-dimensional (3D) reconstruction of a single protein molecule is essential for understanding the relationship between the structural dynamics and functions of the protein. Electron tomography ...Three-dimensional (3D) reconstruction of a single protein molecule is essential for understanding the relationship between the structural dynamics and functions of the protein. Electron tomography (ET) provides a tool for imaging an individual particle of protein from a series of tilted angles. Individual-particle electron tomography (IPET) provides an approach for reconstructing a 3D density map from a single targeted protein particle (without averaging from different particles of this type of protein), in which the target particle was imaged from a series of tilting angles. However, owing to radiation damage limitations, low-dose images (high noise, and low image contrast) are often challenging to be aligned for 3D reconstruction at intermediate resolution (1-3 nm). Here, we propose a computational method to enhance the image contrast, without increasing any experimental dose, for IPET 3D reconstruction. Using an edge-preserving smoothing-based multi-scale image decomposition algorithm, this method can detect the object against a high-noise background and enhance the object image contrast without increasing the noise level or significantly decreasing the image resolution. The method was validated by using both negative staining (NS) ET and cryo-ET images. The successful 3D reconstruction of a small molecule (<100 kDa) indicated that this method can be used as a supporting tool to current ET 3D reconstruction methods for studying protein dynamics via structure determination from each individual particle of the same type of protein. | |||||||||
History |
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-Structure visualization
Movie |
Movie viewer |
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Structure viewer | EM map: SurfViewMolmilJmol/JSmol |
Supplemental images |
-Downloads & links
-EMDB archive
Map data | emd_9269.map.gz | 3.6 MB | EMDB map data format | |
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Header (meta data) | emd-9269-v30.xml emd-9269.xml | 10.9 KB 10.9 KB | Display Display | EMDB header |
FSC (resolution estimation) | emd_9269_fsc.xml | 9.1 KB | Display | FSC data file |
Images | emd_9269.png | 20.9 KB | ||
Archive directory | http://ftp.pdbj.org/pub/emdb/structures/EMD-9269 ftp://ftp.pdbj.org/pub/emdb/structures/EMD-9269 | HTTPS FTP |
-Validation report
Summary document | emd_9269_validation.pdf.gz | 79.2 KB | Display | EMDB validaton report |
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Full document | emd_9269_full_validation.pdf.gz | 78.3 KB | Display | |
Data in XML | emd_9269_validation.xml.gz | 494 B | Display | |
Arichive directory | https://ftp.pdbj.org/pub/emdb/validation_reports/EMD-9269 ftp://ftp.pdbj.org/pub/emdb/validation_reports/EMD-9269 | HTTPS FTP |
-Related structure data
Related structure data | 9262C 9263C 9266C 9267C 9268C 9270C 9271C C: citing same article (ref.) |
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Similar structure data |
-Links
EMDB pages | EMDB (EBI/PDBe) / EMDataResource |
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-Map
File | Download / File: emd_9269.map.gz / Format: CCP4 / Size: 3.8 MB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES) | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
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Projections & slices | Image control
Images are generated by Spider. | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Voxel size | X=Y=Z: 4.8 Å | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Density |
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Symmetry | Space group: 1 | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Details | EMDB XML:
CCP4 map header:
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-Supplemental data
-Sample components
-Entire : Human plasma low-density lipoprotein in complex with cholesteryl ...
Entire | Name: Human plasma low-density lipoprotein in complex with cholesteryl ester transfer protein |
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Components |
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-Supramolecule #1: Human plasma low-density lipoprotein in complex with cholesteryl ...
Supramolecule | Name: Human plasma low-density lipoprotein in complex with cholesteryl ester transfer protein type: complex / ID: 1 / Parent: 0 Details: Low-density lipoproteins (LDLs) were isolated from human plasma by density gradient ultracentrifugation (Children's Hospital Oakland Research Institute). Cholesteryl ester transfer protein ...Details: Low-density lipoproteins (LDLs) were isolated from human plasma by density gradient ultracentrifugation (Children's Hospital Oakland Research Institute). Cholesteryl ester transfer protein (CETP) was produced by MERCK. LDL and CETP were incubated at 37 degC for 15 minutes at a molar ratio of ~4:1. |
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Source (natural) | Organism: Homo sapiens (human) |
Molecular weight | Theoretical: 2.5 MDa |
-Supramolecule #2: Human plasma low-density lipoprotein
Supramolecule | Name: Human plasma low-density lipoprotein / type: complex / ID: 2 / Parent: 1 Details: Low-density lipoproteins (LDLs) were isolated from human plasma by density gradient ultracentrifugation (Children's Hospital Oakland Research Institute). |
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-Supramolecule #3: Cholesteryl ester transfer protein
Supramolecule | Name: Cholesteryl ester transfer protein / type: organelle_or_cellular_component / ID: 3 / Parent: 1 Details: Cholesteryl ester transfer protein (CETP) was produced by MERCK. |
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Source (natural) | Organism: Homo sapiens (human) / Tissue: Blood |
-Experimental details
-Structure determination
Method | cryo EM |
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Processing | electron tomography |
Aggregation state | particle |
-Sample preparation
Buffer | pH: 7 / Details: 1X Dulbeccos phosphate-buffered saline |
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Grid | Model: Homemade / Material: COPPER / Mesh: 200 / Support film - Material: CARBON / Support film - topology: HOLEY / Pretreatment - Type: GLOW DISCHARGE / Pretreatment - Atmosphere: AIR |
Vitrification | Cryogen name: ETHANE / Chamber humidity: 90 % / Instrument: LEICA EM GP |
Details | Low-density lipoproteins (LDLs) were isolated from human plasma by density gradient ultracentrifugation (Children's Hospital Oakland Research Institute). Cholesteryl ester transfer protein (CETP) was produced by MERCK. LDL and CETP were incubated at 37 degC for 15 minutes at a molar ratio of ~4:1. |
Sectioning | Other: NO SECTIONING |
-Electron microscopy
Microscope | ZEISS LIBRA120PLUS |
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Specialist optics | Energy filter - Name: In-column Omega Filter / Energy filter - Slit width: 20 eV |
Image recording | Film or detector model: GATAN ULTRASCAN 4000 (4k x 4k) / Digitization - Dimensions - Width: 4096 pixel / Digitization - Dimensions - Height: 4096 pixel / Average electron dose: 0.26 e/Å2 |
Electron beam | Acceleration voltage: 120 kV / Electron source: LAB6 |
Electron optics | Illumination mode: FLOOD BEAM / Imaging mode: BRIGHT FIELD / Cs: 2.2 mm / Nominal magnification: 50000 |