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Yorodumi- EMDB-30930: The negatively stained reconstruction of cytoplasmic AdeS in ATP-... -
+Open data
-Basic information
Entry | Database: EMDB / ID: EMD-30930 | |||||||||
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Title | The negatively stained reconstruction of cytoplasmic AdeS in ATP-bound state | |||||||||
Map data | AdeS reconstruction in ATP-bound state. | |||||||||
Sample |
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Function / homology | Function and homology information histidine kinase / phosphorelay sensor kinase activity / membrane => GO:0016020 / cytoplasm Similarity search - Function | |||||||||
Biological species | Acinetobacter baumannii (bacteria) | |||||||||
Method | single particle reconstruction / negative staining / Resolution: 22.0 Å | |||||||||
Authors | Zhu L / Wu D / Wu K | |||||||||
Funding support | China, 2 items
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Citation | Journal: iScience / Year: 2021 Title: Proteolysis and multimerization regulate signaling along the two-component regulatory system AdeRS. Authors: Zhenlin Ouyang / Fang Zheng / Li Zhu / Jan Felix / Di Wu / Ke Wu / Irina Gutsche / Yi Wu / Peter M Hwang / Junjun She / Yurong Wen / Abstract: Bacterial two-component regulatory systems are ubiquitous environment-sensing signal transducers involved in pathogenesis and antibiotic resistance. The two-component regulatory system AdeRS is made ...Bacterial two-component regulatory systems are ubiquitous environment-sensing signal transducers involved in pathogenesis and antibiotic resistance. The two-component regulatory system AdeRS is made up of a sensor histidine kinase AdeS and a cognate response regulator AdeR, which together reduce repression of the multidrug-resistant efflux pump AdeABC. Herein we demonstrate that an N-terminal intrinsically disordered tail in AdeR is important for the upregulation of expression, although it greatly increases the susceptibility of AdeR to proteasome-mediated degradation. We also show that AdeS assembles into a hexameric state that is necessary for its full histidine kinase activity, which appears to occur via autophosphorylation. Taken together, this study demonstrates new structural mechanisms through which two-component systems can transduce environmental signals to impact gene expression and enlightens new potential antimicrobial approach by targeting two-component regulatory systems. | |||||||||
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_30930.map.gz | 2.7 MB | EMDB map data format | |
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Header (meta data) | emd-30930-v30.xml emd-30930.xml | 8.9 KB 8.9 KB | Display Display | EMDB header |
FSC (resolution estimation) | emd_30930_fsc.xml | 3.7 KB | Display | FSC data file |
Images | emd_30930.png | 72.2 KB | ||
Archive directory | http://ftp.pdbj.org/pub/emdb/structures/EMD-30930 ftp://ftp.pdbj.org/pub/emdb/structures/EMD-30930 | HTTPS FTP |
-Related structure data
-Links
EMDB pages | EMDB (EBI/PDBe) / EMDataResource |
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Related items in Molecule of the Month |
-Map
File | Download / File: emd_30930.map.gz / Format: CCP4 / Size: 3.8 MB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES) | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
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Annotation | AdeS reconstruction in ATP-bound state. | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Voxel size | X=Y=Z: 3.06 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 : histidine kinase AdeS in ATP-bound state
Entire | Name: histidine kinase AdeS in ATP-bound state |
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Components |
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-Supramolecule #1: histidine kinase AdeS in ATP-bound state
Supramolecule | Name: histidine kinase AdeS in ATP-bound state / type: complex / ID: 1 / Parent: 0 |
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Source (natural) | Organism: Acinetobacter baumannii (bacteria) |
Recombinant expression | Organism: Escherichia coli BL21(DE3) (bacteria) / Recombinant strain: BL21(DE3) |
-Experimental details
-Structure determination
Method | negative staining |
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Processing | single particle reconstruction |
Aggregation state | particle |
-Sample preparation
Concentration | 0.018 mg/mL |
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Buffer | pH: 8 Details: 20mM Tris-HCI (pH8.0), 150mM NaCl, 2mM ATP, 0.1 mM MgCl2 |
Staining | Type: NEGATIVE / Material: uranyl formate |
-Electron microscopy
Microscope | FEI TECNAI F20 |
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Electron beam | Acceleration voltage: 200 kV / Electron source: FIELD EMISSION GUN |
Electron optics | C2 aperture diameter: 100.0 µm / Calibrated defocus max: 1.7 µm / Calibrated defocus min: 0.6 µm / Illumination mode: OTHER / Imaging mode: BRIGHT FIELDBright-field microscopy / Cs: 2.7 mm / Nominal magnification: 92000 |
Image recording | Film or detector model: FEI CETA (4k x 4k) / Number real images: 319 / Average exposure time: 1.0 sec. / Average electron dose: 23.0 e/Å2 |
Experimental equipment | Model: Tecnai F20 / Image courtesy: FEI Company |