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Open data
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Basic information
| Entry | Database: EMDB / ID: EMD-12961 | |||||||||
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| Title | Dps with iron-containing cluster | |||||||||
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| Function / homology | Function and homology informationDnaA-Dps complex / Oxidoreductases; Oxidizing metal ions / oxidoreductase activity, acting on metal ions / nucleoid / chromosome condensation / response to starvation / response to stress / negative regulation of DNA-templated DNA replication initiation / ferric iron binding / intracellular iron ion homeostasis ...DnaA-Dps complex / Oxidoreductases; Oxidizing metal ions / oxidoreductase activity, acting on metal ions / nucleoid / chromosome condensation / response to starvation / response to stress / negative regulation of DNA-templated DNA replication initiation / ferric iron binding / intracellular iron ion homeostasis / DNA binding / identical protein binding / membrane / cytoplasm Similarity search - Function | |||||||||
| Biological species | ![]() | |||||||||
| Method | single particle reconstruction / cryo EM / Resolution: 3.2 Å | |||||||||
Authors | Chesnokov YM / Kamyshinsky RA | |||||||||
| Funding support | Russian Federation, 1 items
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Citation | Journal: Int J Mol Sci / Year: 2021Title: Structural Rearrangement of Dps-DNA Complex Caused by Divalent Mg and Fe Cations. Authors: Liubov Dadinova / Roman Kamyshinsky / Yury Chesnokov / Andrey Mozhaev / Vladimir Matveev / Andrey Gruzinov / Alexander Vasiliev / Eleonora Shtykova / ![]() Abstract: Two independent, complementary methods of structural analysis were used to elucidate the effect of divalent magnesium and iron cations on the structure of the protective Dps-DNA complex. Small-angle ...Two independent, complementary methods of structural analysis were used to elucidate the effect of divalent magnesium and iron cations on the structure of the protective Dps-DNA complex. Small-angle X-ray scattering (SAXS) and cryo-electron microscopy (cryo-EM) demonstrate that Mg ions block the N-terminals of the Dps protein preventing its interaction with DNA. Non-interacting macromolecules of Dps and DNA remain in the solution in this case. The subsequent addition of the chelating agent (EDTA) leads to a complete restoration of the structure of the complex. Different effect was observed when Fe cations were added to the Dps-DNA complex; the presence of Fe in solution leads to the total complex destruction and aggregation without possibility of the complex restoration with the chelating agent. Here, we discuss these different responses of the Dps-DNA complex on the presence of additional free metal cations, investigating the structure of the Dps protein with and without cations using SAXS and cryo-EM. Additionally, the single particle analysis of Dps with accumulated iron performed by cryo-EM shows localization of iron nanoparticles inside the Dps cavity next to the acidic (hydrophobic) pore, near three glutamate residues. | |||||||||
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Structure visualization
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| Structure viewer | EM map: SurfView Molmil Jmol/JSmol |
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Downloads & links
-EMDB archive
| Map data | emd_12961.map.gz | 5.2 MB | EMDB map data format | |
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| Header (meta data) | emd-12961-v30.xml emd-12961.xml | 18.1 KB 18.1 KB | Display Display | EMDB header |
| FSC (resolution estimation) | emd_12961_fsc.xml | 8.8 KB | Display | FSC data file |
| Images | emd_12961.png | 238.8 KB | ||
| Masks | emd_12961_msk_1.map | 27 MB | Mask map | |
| Others | emd_12961_half_map_1.map.gz emd_12961_half_map_2.map.gz | 20.7 MB 20.6 MB | ||
| Archive directory | http://ftp.pdbj.org/pub/emdb/structures/EMD-12961 ftp://ftp.pdbj.org/pub/emdb/structures/EMD-12961 | HTTPS FTP |
-Validation report
| Summary document | emd_12961_validation.pdf.gz | 386.2 KB | Display | EMDB validaton report |
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| Full document | emd_12961_full_validation.pdf.gz | 385.3 KB | Display | |
| Data in XML | emd_12961_validation.xml.gz | 13.8 KB | Display | |
| Data in CIF | emd_12961_validation.cif.gz | 17.6 KB | Display | |
| Arichive directory | https://ftp.pdbj.org/pub/emdb/validation_reports/EMD-12961 ftp://ftp.pdbj.org/pub/emdb/validation_reports/EMD-12961 | HTTPS FTP |
-Related structure data
| Similar structure data |
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Links
| EMDB pages | EMDB (EBI/PDBe) / EMDataResource |
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| Related items in Molecule of the Month |
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Map
| File | Download / File: emd_12961.map.gz / Format: CCP4 / Size: 27 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: 0.86 Å | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
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| Symmetry | Space group: 1 | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
| Details | EMDB XML:
CCP4 map header:
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-Supplemental data
-Mask #1
| File | emd_12961_msk_1.map | ||||||||||||
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-Half map: half2
| File | emd_12961_half_map_1.map | ||||||||||||
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| Annotation | half2 | ||||||||||||
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| Density Histograms |
-Half map: half1
| File | emd_12961_half_map_2.map | ||||||||||||
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| Annotation | half1 | ||||||||||||
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Sample components
-Entire : Dps with iron-containing cluster
| Entire | Name: Dps with iron-containing cluster |
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| Components |
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-Supramolecule #1: Dps with iron-containing cluster
| Supramolecule | Name: Dps with iron-containing cluster / type: complex / ID: 1 / Parent: 0 |
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| Source (natural) | Organism: ![]() |
| Recombinant expression | Organism: ![]() |
| Molecular weight | Theoretical: 224 KDa |
-Experimental details
-Structure determination
| Method | cryo EM |
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Processing | single particle reconstruction |
| Aggregation state | particle |
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Sample preparation
| Concentration | 3.5 mg/mL | ||||||||||
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| Buffer | pH: 8 Component:
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| Grid | Model: Quantifoil R1.2/1.3 / Material: COPPER / Mesh: 300 / Pretreatment - Type: GLOW DISCHARGE / Pretreatment - Atmosphere: AIR / Pretreatment - Pressure: 0.026000000000000002 kPa / Details: Pelco EasiGlow ( 25 mA) | ||||||||||
| Vitrification | Cryogen name: ETHANE / Chamber humidity: 100 % / Chamber temperature: 293 K / Instrument: FEI VITROBOT MARK IV / Details: blotting for 2.5 seconds. | ||||||||||
| Details | 50 ul of Dps (3.5 mg/mL) were mixed with 1 ul of 1M FeSO4. |
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Electron microscopy
| Microscope | FEI TITAN KRIOS |
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| Temperature | Min: 78.0 K / Max: 90.0 K |
| Specialist optics | Spherical aberration corrector: Cs corrector by CEOS |
| Image recording | Film or detector model: FEI FALCON II (4k x 4k) / Detector mode: INTEGRATING / Digitization - Dimensions - Width: 4096 pixel / Digitization - Dimensions - Height: 4096 pixel / Digitization - Sampling interval: 14.0 µm / Digitization - Frames/image: 1-40 / Number grids imaged: 1 / Number real images: 129 / Average exposure time: 2.0 sec. / Average electron dose: 80.0 e/Å2 |
| Electron beam | Acceleration voltage: 300 kV / Electron source: FIELD EMISSION GUN |
| Electron optics | C2 aperture diameter: 100.0 µm / Calibrated defocus max: 2.5 µm / Calibrated defocus min: 0.8 µm / Calibrated magnification: 162790 / Illumination mode: FLOOD BEAM / Imaging mode: BRIGHT FIELD / Cs: 0.005 mm / Nominal defocus max: 2.5 µm / Nominal defocus min: 0.8 µm / Nominal magnification: 75000 |
| Sample stage | Specimen holder model: FEI TITAN KRIOS AUTOGRID HOLDER / Cooling holder cryogen: NITROGEN |
| Experimental equipment | ![]() Model: Titan Krios / Image courtesy: FEI Company |
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Image processing
-Atomic model buiding 1
| Initial model | PDB ID: |
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| Refinement | Space: REAL / Protocol: RIGID BODY FIT |
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Authors
Russian Federation, 1 items
Citation
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