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Yorodumi- EMDB-15905: Cryo-EM structure of the E.coli 70S ribosome in complex with the ... -
+Open data
-Basic information
Entry | Database: EMDB / ID: EMD-15905 | |||||||||
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Title | Cryo-EM structure of the E.coli 70S ribosome in complex with the antibiotic Myxovalargin B. | |||||||||
Map data | Post-processed masked map | |||||||||
Sample |
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Function / homology | Function and homology information ornithine decarboxylase inhibitor activity / transcription antitermination factor activity, RNA binding / misfolded RNA binding / Group I intron splicing / RNA folding / transcriptional attenuation / endoribonuclease inhibitor activity / RNA-binding transcription regulator activity / positive regulation of ribosome biogenesis / negative regulation of cytoplasmic translation ...ornithine decarboxylase inhibitor activity / transcription antitermination factor activity, RNA binding / misfolded RNA binding / Group I intron splicing / RNA folding / transcriptional attenuation / endoribonuclease inhibitor activity / RNA-binding transcription regulator activity / positive regulation of ribosome biogenesis / negative regulation of cytoplasmic translation / four-way junction DNA binding / DnaA-L2 complex / translation repressor activity / negative regulation of DNA-templated DNA replication initiation / negative regulation of translational initiation / regulation of mRNA stability / mRNA regulatory element binding translation repressor activity / ribosome assembly / assembly of large subunit precursor of preribosome / positive regulation of RNA splicing / transcription elongation factor complex / cytosolic ribosome assembly / regulation of DNA-templated transcription elongation / DNA endonuclease activity / response to reactive oxygen species / transcription antitermination / regulation of cell growth / DNA-templated transcription termination / maintenance of translational fidelity / response to radiation / mRNA 5'-UTR binding / ribosomal small subunit biogenesis / small ribosomal subunit rRNA binding / large ribosomal subunit / ribosome biogenesis / ribosome binding / regulation of translation / ribosomal small subunit assembly / small ribosomal subunit / 5S rRNA binding / large ribosomal subunit rRNA binding / transferase activity / cytosolic small ribosomal subunit / ribosomal large subunit assembly / cytoplasmic translation / cytosolic large ribosomal subunit / tRNA binding / molecular adaptor activity / negative regulation of translation / rRNA binding / ribosome / structural constituent of ribosome / translation / response to antibiotic / negative regulation of DNA-templated transcription / mRNA binding / DNA binding / RNA binding / zinc ion binding / membrane / cytosol / cytoplasm Similarity search - Function | |||||||||
Biological species | Escherichia coli BW25113 (bacteria) / Escherichia coli K-12 (bacteria) / Myxococcus fulvus (bacteria) | |||||||||
Method | single particle reconstruction / cryo EM / Resolution: 3.0 Å | |||||||||
Authors | Koller TO / Graf M / Wilson DN | |||||||||
Funding support | Germany, 1 items
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Citation | Journal: J Am Chem Soc / Year: 2023 Title: The Myxobacterial Antibiotic Myxovalargin: Biosynthesis, Structural Revision, Total Synthesis, and Molecular Characterization of Ribosomal Inhibition. Authors: Timm O Koller / Ullrich Scheid / Teresa Kösel / Jennifer Herrmann / Daniel Krug / Helena I M Boshoff / Bertrand Beckert / Joanna C Evans / Jan Schlemmer / Becky Sloan / Danielle M Weiner / ...Authors: Timm O Koller / Ullrich Scheid / Teresa Kösel / Jennifer Herrmann / Daniel Krug / Helena I M Boshoff / Bertrand Beckert / Joanna C Evans / Jan Schlemmer / Becky Sloan / Danielle M Weiner / Laura E Via / Atica Moosa / Thomas R Ioerger / Michael Graf / Boris Zinshteyn / Maha Abdelshahid / Fabian Nguyen / Stefan Arenz / Franziska Gille / Maik Siebke / Tim Seedorf / Oliver Plettenburg / Rachel Green / Anna-Luisa Warnke / Joachim Ullrich / Ralf Warrass / Clifton E Barry / Digby F Warner / Valerie Mizrahi / Andreas Kirschning / Daniel N Wilson / Rolf Müller / Abstract: Resistance of bacterial pathogens against antibiotics is declared by WHO as a major global health threat. As novel antibacterial agents are urgently needed, we re-assessed the broad-spectrum ...Resistance of bacterial pathogens against antibiotics is declared by WHO as a major global health threat. As novel antibacterial agents are urgently needed, we re-assessed the broad-spectrum myxobacterial antibiotic myxovalargin and found it to be extremely potent against . To ensure compound supply for further development, we studied myxovalargin biosynthesis in detail enabling production via fermentation of a native producer. Feeding experiments as well as functional genomics analysis suggested a structural revision, which was eventually corroborated by the development of a concise total synthesis. The ribosome was identified as the molecular target based on resistant mutant sequencing, and a cryo-EM structure revealed that myxovalargin binds within and completely occludes the exit tunnel, consistent with a mode of action to arrest translation during a late stage of translation initiation. These studies open avenues for structure-based scaffold improvement toward development as an antibacterial agent. | |||||||||
History |
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-Structure visualization
Supplemental images |
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-Downloads & links
-EMDB archive
Map data | emd_15905.map.gz | 25 MB | EMDB map data format | |
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Header (meta data) | emd-15905-v30.xml emd-15905.xml | 70.2 KB 70.2 KB | Display Display | EMDB header |
FSC (resolution estimation) | emd_15905_fsc.xml | 12.7 KB | Display | FSC data file |
Images | emd_15905.png | 87.6 KB | ||
Others | emd_15905_additional_1.map.gz emd_15905_additional_2.map.gz emd_15905_half_map_1.map.gz emd_15905_half_map_2.map.gz | 166.1 MB 138.9 MB 140.5 MB 140.5 MB | ||
Archive directory | http://ftp.pdbj.org/pub/emdb/structures/EMD-15905 ftp://ftp.pdbj.org/pub/emdb/structures/EMD-15905 | HTTPS FTP |
-Validation report
Summary document | emd_15905_validation.pdf.gz | 769.3 KB | Display | EMDB validaton report |
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Full document | emd_15905_full_validation.pdf.gz | 768.9 KB | Display | |
Data in XML | emd_15905_validation.xml.gz | 20.1 KB | Display | |
Data in CIF | emd_15905_validation.cif.gz | 26.8 KB | Display | |
Arichive directory | https://ftp.pdbj.org/pub/emdb/validation_reports/EMD-15905 ftp://ftp.pdbj.org/pub/emdb/validation_reports/EMD-15905 | HTTPS FTP |
-Related structure data
Related structure data | 8b7yMC 7qq3C C: citing same article (ref.) M: atomic model generated by this map |
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Similar structure data | Similarity search - Function & homologyF&H Search |
-Links
EMDB pages | EMDB (EBI/PDBe) / EMDataResource |
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Related items in Molecule of the Month |
-Map
File | Download / File: emd_15905.map.gz / Format: CCP4 / Size: 178 MB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES) | ||||||||||||||||||||||||||||||||||||
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Annotation | Post-processed masked map | ||||||||||||||||||||||||||||||||||||
Projections & slices | Image control
Images are generated by Spider. | ||||||||||||||||||||||||||||||||||||
Voxel size | X=Y=Z: 1.053 Å | ||||||||||||||||||||||||||||||||||||
Density |
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Symmetry | Space group: 1 | ||||||||||||||||||||||||||||||||||||
Details | EMDB XML:
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-Supplemental data
-Additional map: post-processed map
File | emd_15905_additional_1.map | ||||||||||||
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Annotation | post-processed map | ||||||||||||
Projections & Slices |
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Density Histograms |
-Additional map: 3D refined map
File | emd_15905_additional_2.map | ||||||||||||
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Annotation | 3D refined map | ||||||||||||
Projections & Slices |
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Density Histograms |
-Half map: Half-map2
File | emd_15905_half_map_1.map | ||||||||||||
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Annotation | Half-map2 | ||||||||||||
Projections & Slices |
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Density Histograms |
-Half map: Half-map1
File | emd_15905_half_map_2.map | ||||||||||||
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Annotation | Half-map1 | ||||||||||||
Projections & Slices |
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Density Histograms |
-Sample components
+Entire : Cryo-EM structure of the E.coli 70S ribosome in complex with the ...
+Supramolecule #1: Cryo-EM structure of the E.coli 70S ribosome in complex with the ...
+Macromolecule #1: 23S ribosomal RNA
+Macromolecule #2: 5S ribosomal RNA
+Macromolecule #29: P-tRNA fMet
+Macromolecule #30: 16S ribosomal RNA
+Macromolecule #51: messenger RNA
+Macromolecule #3: 50S ribosomal protein L2
+Macromolecule #4: 50S ribosomal protein L3
+Macromolecule #5: 50S ribosomal protein L4
+Macromolecule #6: 50S ribosomal protein L6
+Macromolecule #7: 50S ribosomal protein L13
+Macromolecule #8: 50S ribosomal protein L14
+Macromolecule #9: 50S ribosomal protein L15
+Macromolecule #10: 50S ribosomal protein L16
+Macromolecule #11: 50S ribosomal protein L17
+Macromolecule #12: 50S ribosomal protein L18
+Macromolecule #13: 50S ribosomal protein L19
+Macromolecule #14: 50S ribosomal protein L20
+Macromolecule #15: 50S ribosomal protein L21
+Macromolecule #16: 50S ribosomal protein L22
+Macromolecule #17: 50S ribosomal protein L23
+Macromolecule #18: 50S ribosomal protein L24
+Macromolecule #19: 50S ribosomal protein L25
+Macromolecule #20: 50S ribosomal protein L27
+Macromolecule #21: 50S ribosomal protein L28
+Macromolecule #22: 50S ribosomal protein L29
+Macromolecule #23: 50S ribosomal protein L30
+Macromolecule #24: 50S ribosomal protein L32
+Macromolecule #25: 50S ribosomal protein L33
+Macromolecule #26: 50S ribosomal protein L34
+Macromolecule #27: 50S ribosomal protein L35
+Macromolecule #28: 50S ribosomal protein L36
+Macromolecule #31: 30S ribosomal protein S2
+Macromolecule #32: 30S ribosomal protein S3
+Macromolecule #33: 30S ribosomal protein S4
+Macromolecule #34: 30S ribosomal protein S5
+Macromolecule #35: 30S ribosomal protein S6
+Macromolecule #36: 30S ribosomal protein S7
+Macromolecule #37: 30S ribosomal protein S8
+Macromolecule #38: 30S ribosomal protein S9
+Macromolecule #39: 30S ribosomal protein S10
+Macromolecule #40: 30S ribosomal protein S11
+Macromolecule #41: 30S ribosomal protein S12
+Macromolecule #42: 30S ribosomal protein S13
+Macromolecule #43: 30S ribosomal protein S14
+Macromolecule #44: 30S ribosomal protein S15
+Macromolecule #45: 30S ribosomal protein S16
+Macromolecule #46: 30S ribosomal protein S17
+Macromolecule #47: 30S ribosomal protein S18
+Macromolecule #48: 30S ribosomal protein S19
+Macromolecule #49: 30S ribosomal protein S20
+Macromolecule #50: 30S ribosomal protein S21
+Macromolecule #52: Myxovalargin B
+Macromolecule #53: MAGNESIUM ION
+Macromolecule #54: ZINC ION
+Macromolecule #55: N-FORMYLMETHIONINE
+Macromolecule #56: SPERMIDINE
+Macromolecule #57: water
-Experimental details
-Structure determination
Method | cryo EM |
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Processing | single particle reconstruction |
Aggregation state | particle |
-Sample preparation
Buffer | pH: 7.4 |
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Grid | Model: Quantifoil R2/2 / Material: COPPER / Mesh: 200 / Support film - Material: CARBON / Support film - topology: HOLEY / Support film - Film thickness: 20.0 nm |
Vitrification | Cryogen name: ETHANE-PROPANE / Chamber humidity: 100 % / Chamber temperature: 277 K / Instrument: FEI VITROBOT MARK IV |
-Electron microscopy
Microscope | FEI TITAN KRIOS |
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Image recording | Film or detector model: FEI FALCON II (4k x 4k) / Detector mode: COUNTING / Average electron dose: 2.5 e/Å2 |
Electron beam | Acceleration voltage: 300 kV / Electron source: FIELD EMISSION GUN |
Electron optics | Illumination mode: FLOOD BEAM / Imaging mode: BRIGHT FIELD / Nominal defocus max: 1.2 µm / Nominal defocus min: 0.7000000000000001 µm |
Experimental equipment | Model: Titan Krios / Image courtesy: FEI Company |