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Yorodumi- EMDB-26959: Cutibacterium acnes 70S ribosome with mRNA, P-site tRNA and Sarec... -
+Open data
-Basic information
Entry | Database: EMDB / ID: EMD-26959 | |||||||||
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Title | Cutibacterium acnes 70S ribosome with mRNA, P-site tRNA and Sarecycline bound | |||||||||
Map data | 70S ribosome with mRNA, P-site tRNA and Sarecycline bound | |||||||||
Sample |
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Keywords | ribosome / sarecycline / acne / ANTIBIOTIC | |||||||||
Function / homology | Function and homology information small ribosomal subunit rRNA binding / large ribosomal subunit / ribosomal small subunit assembly / large ribosomal subunit rRNA binding / small ribosomal subunit / 5S rRNA binding / transferase activity / cytosolic small ribosomal subunit / ribosomal large subunit assembly / cytoplasmic translation ...small ribosomal subunit rRNA binding / large ribosomal subunit / ribosomal small subunit assembly / large ribosomal subunit rRNA binding / small ribosomal subunit / 5S rRNA binding / transferase activity / cytosolic small ribosomal subunit / ribosomal large subunit assembly / cytoplasmic translation / cytosolic large ribosomal subunit / tRNA binding / negative regulation of translation / rRNA binding / ribosome / structural constituent of ribosome / translation / ribonucleoprotein complex / mRNA binding / zinc ion binding / metal ion binding / cytosol / cytoplasm Similarity search - Function | |||||||||
Biological species | Cutibacterium acnes (bacteria) / Escherichia coli (E. coli) | |||||||||
Method | single particle reconstruction / cryo EM / Resolution: 2.78 Å | |||||||||
Authors | Lomakin IB / Devarkar SC / Bunick CG | |||||||||
Funding support | United States, 1 items
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Citation | Journal: Nucleic Acids Res / Year: 2023 Title: Sarecycline inhibits protein translation in Cutibacterium acnes 70S ribosome using a two-site mechanism. Authors: Ivan B Lomakin / Swapnil C Devarkar / Shivali Patel / Ayman Grada / Christopher G Bunick / Abstract: Acne vulgaris is a chronic disfiguring skin disease affecting ∼1 billion people worldwide, often having persistent negative effects on physical and mental health. The Gram-positive anaerobe, ...Acne vulgaris is a chronic disfiguring skin disease affecting ∼1 billion people worldwide, often having persistent negative effects on physical and mental health. The Gram-positive anaerobe, Cutibacterium acnes is implicated in acne pathogenesis and is, therefore, a main target for antibiotic-based acne therapy. We determined a 2.8-Å resolution structure of the 70S ribosome of Cutibacterium acnes by cryogenic electron microscopy and discovered that sarecycline, a narrow-spectrum antibiotic against Cutibacterium acnes, may inhibit two active sites of this bacterium's ribosome in contrast to the one site detected previously on the model ribosome of Thermus thermophilus. Apart from the canonical binding site at the mRNA decoding center, the second binding site for sarecycline exists at the nascent peptide exit tunnel, reminiscent of the macrolides class of antibiotics. The structure also revealed Cutibacterium acnes-specific features of the ribosomal RNA and proteins. Unlike the ribosome of the Gram-negative bacterium Escherichia coli, Cutibacterium acnes ribosome has two additional proteins, bS22 and bL37, which are also present in the ribosomes of Mycobacterium smegmatis and Mycobacterium tuberculosis. We show that bS22 and bL37 have antimicrobial properties and may be involved in maintaining the healthy homeostasis of the human skin microbiome. | |||||||||
History |
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-Structure visualization
Supplemental images |
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-Downloads & links
-EMDB archive
Map data | emd_26959.map.gz | 76.1 MB | EMDB map data format | |
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Header (meta data) | emd-26959-v30.xml emd-26959.xml | 72 KB 72 KB | Display Display | EMDB header |
FSC (resolution estimation) | emd_26959_fsc.xml | 12.6 KB | Display | FSC data file |
Images | emd_26959.png | 84.4 KB | ||
Filedesc metadata | emd-26959.cif.gz | 14.6 KB | ||
Others | emd_26959_half_map_1.map.gz emd_26959_half_map_2.map.gz | 139.4 MB 139.4 MB | ||
Archive directory | http://ftp.pdbj.org/pub/emdb/structures/EMD-26959 ftp://ftp.pdbj.org/pub/emdb/structures/EMD-26959 | HTTPS FTP |
-Validation report
Summary document | emd_26959_validation.pdf.gz | 1.2 MB | Display | EMDB validaton report |
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Full document | emd_26959_full_validation.pdf.gz | 1.2 MB | Display | |
Data in XML | emd_26959_validation.xml.gz | 20.1 KB | Display | |
Data in CIF | emd_26959_validation.cif.gz | 26 KB | Display | |
Arichive directory | https://ftp.pdbj.org/pub/emdb/validation_reports/EMD-26959 ftp://ftp.pdbj.org/pub/emdb/validation_reports/EMD-26959 | HTTPS FTP |
-Related structure data
Related structure data | 8crxMC 8cvmC 8cvoC 8cwoC M: atomic model generated by this map C: citing same article (ref.) |
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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_26959.map.gz / Format: CCP4 / Size: 149.9 MB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES) | ||||||||||||||||||||||||||||||||||||
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Annotation | 70S ribosome with mRNA, P-site tRNA and Sarecycline bound | ||||||||||||||||||||||||||||||||||||
Projections & slices | Image control
Images are generated by Spider. | ||||||||||||||||||||||||||||||||||||
Voxel size | X=Y=Z: 1.068 Å | ||||||||||||||||||||||||||||||||||||
Density |
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Symmetry | Space group: 1 | ||||||||||||||||||||||||||||||||||||
Details | EMDB XML:
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-Supplemental data
-Half map: 70S ribosome with mRNA, P-site tRNA and Sarecycline bound
File | emd_26959_half_map_1.map | ||||||||||||
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Annotation | 70S ribosome with mRNA, P-site tRNA and Sarecycline bound | ||||||||||||
Projections & Slices |
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Density Histograms |
-Half map: 70S ribosome with mRNA, P-site tRNA and Sarecycline bound
File | emd_26959_half_map_2.map | ||||||||||||
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Annotation | 70S ribosome with mRNA, P-site tRNA and Sarecycline bound | ||||||||||||
Projections & Slices |
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Density Histograms |
-Sample components
+Entire : 70S ribosome with mRNA, P-site tRNA and Sarecycline bound
+Supramolecule #1: 70S ribosome with mRNA, P-site tRNA and Sarecycline bound
+Macromolecule #1: RNA (1537-MER)
+Macromolecule #49: 23S ribosomal RNA
+Macromolecule #50: 5S ribosomal RNA
+Macromolecule #52: 32MF mRNA
+Macromolecule #53: P-site initiator tRNA
+Macromolecule #2: 30S ribosomal protein S4
+Macromolecule #3: 30S ribosomal protein S5
+Macromolecule #4: 30S ribosomal protein S6
+Macromolecule #5: 30S ribosomal protein S8
+Macromolecule #6: 30S ribosomal protein S11
+Macromolecule #7: 30S ribosomal protein S12
+Macromolecule #8: 30S ribosomal protein S15
+Macromolecule #9: 30S ribosomal protein S17
+Macromolecule #10: 30S ribosomal protein S18
+Macromolecule #11: 30S ribosomal protein S20
+Macromolecule #12: 30S ribosomal protein S16
+Macromolecule #13: 30S ribosomal protein bS22
+Macromolecule #14: 30S ribosomal protein S3
+Macromolecule #15: 30S ribosomal protein S7
+Macromolecule #16: 30S ribosomal protein S9
+Macromolecule #17: 30S ribosomal protein S10
+Macromolecule #18: 30S ribosomal protein S13
+Macromolecule #19: 30S ribosomal protein S14 type Z
+Macromolecule #20: 30S ribosomal protein S19
+Macromolecule #21: 30S ribosomal protein S2
+Macromolecule #22: 50S ribosomal protein L2
+Macromolecule #23: 50S ribosomal protein L3
+Macromolecule #24: 50S ribosomal protein L4
+Macromolecule #25: 50S ribosomal protein L5
+Macromolecule #26: 50S ribosomal protein L6
+Macromolecule #27: 50S ribosomal protein L13
+Macromolecule #28: 50S ribosomal protein L14
+Macromolecule #29: 50S ribosomal protein L15
+Macromolecule #30: 50S ribosomal protein L16
+Macromolecule #31: 50S ribosomal protein L17
+Macromolecule #32: 50S ribosomal protein L18
+Macromolecule #33: 50S ribosomal protein L19
+Macromolecule #34: 50S ribosomal protein L20
+Macromolecule #35: 50S ribosomal protein L21
+Macromolecule #36: 50S ribosomal protein L22
+Macromolecule #37: 50S ribosomal protein L23
+Macromolecule #38: 50S ribosomal protein L24
+Macromolecule #39: 50S ribosomal protein L25
+Macromolecule #40: 50S ribosomal protein L27
+Macromolecule #41: 50S ribosomal protein L28
+Macromolecule #42: 50S ribosomal protein L29
+Macromolecule #43: 50S ribosomal protein L30
+Macromolecule #44: 50S ribosomal protein L32
+Macromolecule #45: 50S ribosomal protein L33
+Macromolecule #46: 50S ribosomal protein L34
+Macromolecule #47: 50S ribosomal protein L35
+Macromolecule #48: 50S ribosomal protein L31
+Macromolecule #51: 50S ribosomal protein bL37
+Macromolecule #54: 50S ribosomal protein L36
+Macromolecule #55: Sarecycline
+Macromolecule #56: MAGNESIUM ION
+Macromolecule #57: ZINC ION
+Macromolecule #58: 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: C-flat-2/1 / Material: GOLD / Mesh: 300 / Pretreatment - Type: GLOW DISCHARGE / Pretreatment - Time: 15 sec. / Details: 25 mA |
Vitrification | Cryogen name: ETHANE / Chamber humidity: 100 % / Instrument: FEI VITROBOT MARK IV |
-Electron microscopy
Microscope | FEI TITAN KRIOS |
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Image recording | Film or detector model: GATAN K3 (6k x 4k) / Average exposure time: 2.1 sec. / Average electron dose: 30.56 e/Å2 |
Electron beam | Acceleration voltage: 300 kV / Electron source: FIELD EMISSION GUN |
Electron optics | C2 aperture diameter: 50.0 µm / Illumination mode: FLOOD BEAM / Imaging mode: BRIGHT FIELD / Cs: 2.7 mm / Nominal defocus max: 2.0 µm / Nominal defocus min: 0.5 µm |
Sample stage | Cooling holder cryogen: NITROGEN |
Experimental equipment | Model: Titan Krios / Image courtesy: FEI Company |
+Image processing
-Atomic model buiding 1
Refinement | Space: REAL / Protocol: FLEXIBLE FIT |
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Output model | PDB-8crx: |