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Yorodumi- EMDB-29786: Structure of WT E.coli 70S ribosome complexed with mRNA, P-site f... -
+Open data
-Basic information
Entry | Database: EMDB / ID: EMD-29786 | |||||||||
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Title | Structure of WT E.coli 70S ribosome complexed with mRNA, P-site fMet-NH-tRNAfMet and A-site ortho-aminobenzoic acid charged NH-tRNAPhe | |||||||||
Map data | 70S global refined map | |||||||||
Sample |
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Keywords | ribosome / non-natural monomers / aminobenzoic acids / unnatural monomers | |||||||||
Function / homology | Function and homology information negative regulation of cytoplasmic translational initiation / positive regulation of ribosome biogenesis / DnaA-L2 complex / negative regulation of DNA-templated DNA replication initiation / negative regulation of translational initiation / mRNA regulatory element binding translation repressor activity / ribosome assembly / assembly of large subunit precursor of preribosome / cytosolic ribosome assembly / transcription antitermination ...negative regulation of cytoplasmic translational initiation / positive regulation of ribosome biogenesis / DnaA-L2 complex / negative regulation of DNA-templated DNA replication initiation / negative regulation of translational initiation / mRNA regulatory element binding translation repressor activity / ribosome assembly / assembly of large subunit precursor of preribosome / cytosolic ribosome assembly / transcription antitermination / regulation of cell growth / translational initiation / DNA-templated transcription termination / maintenance of translational fidelity / mRNA 5'-UTR binding / ribosomal small subunit biogenesis / small ribosomal subunit rRNA binding / large ribosomal subunit / ribosome binding / 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 / negative regulation of translation / rRNA binding / ribosome / structural constituent of ribosome / ribonucleoprotein complex / translation / response to antibiotic / mRNA binding / RNA binding / zinc ion binding / membrane / cytosol / cytoplasm Similarity search - Function | |||||||||
Biological species | Escherichia coli (E. coli) | |||||||||
Method | single particle reconstruction / cryo EM / Resolution: 2.02 Å | |||||||||
Authors | Majumdar C / Cate JHD | |||||||||
Funding support | United States, 1 items
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Citation | Journal: ACS Cent Sci / Year: 2023 Title: Aminobenzoic Acid Derivatives Obstruct Induced Fit in the Catalytic Center of the Ribosome. Authors: Chandrima Majumdar / Joshua A Walker / Matthew B Francis / Alanna Schepartz / Jamie H D Cate / Abstract: The () ribosome can incorporate a variety of non-l-α-amino acid monomers into polypeptide chains but with poor efficiency. Although these monomers span a diverse set of compounds, there exists no ...The () ribosome can incorporate a variety of non-l-α-amino acid monomers into polypeptide chains but with poor efficiency. Although these monomers span a diverse set of compounds, there exists no high-resolution structural information regarding their positioning within the catalytic center of the ribosome, the peptidyl transferase center (PTC). Thus, details regarding the mechanism of amide bond formation and the structural basis for differences and defects in incorporation efficiency remain unknown. Within a set of three aminobenzoic acid derivatives-3-aminopyridine-4-carboxylic acid (Apy), aminobenzoic acid (ABZ), and aminobenzoic acid (ABZ)-the ribosome incorporates Apy into polypeptide chains with the highest efficiency, followed by ABZ and then ABZ, a trend that does not track with the nucleophilicity of the reactive amines. Here, we report high-resolution cryo-EM structures of the ribosome with each of these three aminobenzoic acid derivatives charged on tRNA bound in the aminoacyl-tRNA site (A-site). The structures reveal how the aromatic ring of each monomer sterically blocks the positioning of nucleotide U2506, thereby preventing rearrangement of nucleotide U2585 and the resulting induced fit in the PTC required for efficient amide bond formation. They also reveal disruptions to the bound water network that is believed to facilitate formation and breakdown of the tetrahedral intermediate. Together, the cryo-EM structures reported here provide a mechanistic rationale for differences in reactivity of aminobenzoic acid derivatives relative to l-α-amino acids and each other and identify stereochemical constraints on the size and geometry of non-monomers that can be accepted efficiently by wild-type ribosomes. | |||||||||
History |
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-Structure visualization
Supplemental images |
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-Downloads & links
-EMDB archive
Map data | emd_29786.map.gz | 46 MB | EMDB map data format | |
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Header (meta data) | emd-29786-v30.xml emd-29786.xml | 74.2 KB 74.2 KB | Display Display | EMDB header |
FSC (resolution estimation) | emd_29786_fsc.xml | 16.3 KB | Display | FSC data file |
Images | emd_29786.png | 89.1 KB | ||
Filedesc metadata | emd-29786.cif.gz | 14.8 KB | ||
Others | emd_29786_half_map_1.map.gz emd_29786_half_map_2.map.gz | 305.1 MB 81.9 MB | ||
Archive directory | http://ftp.pdbj.org/pub/emdb/structures/EMD-29786 ftp://ftp.pdbj.org/pub/emdb/structures/EMD-29786 | HTTPS FTP |
-Validation report
Summary document | emd_29786_validation.pdf.gz | 866.7 KB | Display | EMDB validaton report |
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Full document | emd_29786_full_validation.pdf.gz | 866.2 KB | Display | |
Data in XML | emd_29786_validation.xml.gz | 24.2 KB | Display | |
Data in CIF | emd_29786_validation.cif.gz | 32.1 KB | Display | |
Arichive directory | https://ftp.pdbj.org/pub/emdb/validation_reports/EMD-29786 ftp://ftp.pdbj.org/pub/emdb/validation_reports/EMD-29786 | HTTPS FTP |
-Related structure data
Related structure data | 8g6wMC 8g6xC 8g6yC 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_29786.map.gz / Format: CCP4 / Size: 381.1 MB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES) | ||||||||||||||||||||||||||||||||||||
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Annotation | 70S global refined map | ||||||||||||||||||||||||||||||||||||
Projections & slices | Image control
Images are generated by Spider. | ||||||||||||||||||||||||||||||||||||
Voxel size | X=Y=Z: 0.8232 Å | ||||||||||||||||||||||||||||||||||||
Density |
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Symmetry | Space group: 1 | ||||||||||||||||||||||||||||||||||||
Details | EMDB XML:
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-Supplemental data
-Half map: 70S global refinement half map
File | emd_29786_half_map_1.map | ||||||||||||
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Annotation | 70S global refinement half map | ||||||||||||
Projections & Slices |
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Density Histograms |
-Half map: 70S global refinement half map
File | emd_29786_half_map_2.map | ||||||||||||
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Annotation | 70S global refinement half map | ||||||||||||
Projections & Slices |
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Density Histograms |
-Sample components
+Entire : 70S E.coli ribosome
+Supramolecule #1: 70S E.coli ribosome
+Macromolecule #1: 50S ribosomal protein L33
+Macromolecule #2: 50S ribosomal protein L34
+Macromolecule #3: 50S ribosomal protein L35
+Macromolecule #4: 50S ribosomal protein L36
+Macromolecule #5: 50S ribosomal protein L31
+Macromolecule #7: 30S ribosomal protein S2
+Macromolecule #8: 30S ribosomal protein S3
+Macromolecule #9: 30S ribosomal protein S4
+Macromolecule #10: 30S ribosomal protein S5
+Macromolecule #11: 30S ribosomal protein S6
+Macromolecule #12: 30S ribosomal protein S7
+Macromolecule #13: 30S ribosomal protein S8
+Macromolecule #14: 30S ribosomal protein S9
+Macromolecule #15: 30S ribosomal protein S10
+Macromolecule #16: 30S ribosomal protein S11
+Macromolecule #17: 30S ribosomal protein S12
+Macromolecule #18: 30S ribosomal protein S13
+Macromolecule #19: 30S ribosomal protein S14
+Macromolecule #20: 30S ribosomal protein S15
+Macromolecule #21: 30S ribosomal protein S16
+Macromolecule #22: 30S ribosomal protein S17
+Macromolecule #23: 30S ribosomal protein S18
+Macromolecule #24: 30S ribosomal protein S19
+Macromolecule #25: 30S ribosomal protein S20
+Macromolecule #26: 30S ribosomal protein S21
+Macromolecule #32: 50S ribosomal protein L2
+Macromolecule #33: 50S ribosomal protein L3
+Macromolecule #34: 50S ribosomal protein L4
+Macromolecule #35: 50S ribosomal protein L5
+Macromolecule #36: 50S ribosomal protein L6
+Macromolecule #37: 50S ribosomal protein L9
+Macromolecule #38: 50S ribosomal protein L13
+Macromolecule #39: 50S ribosomal protein L14
+Macromolecule #40: 50S ribosomal protein L15
+Macromolecule #41: 50S ribosomal protein L16
+Macromolecule #42: 50S ribosomal protein L17
+Macromolecule #43: 50S ribosomal protein L18
+Macromolecule #44: 50S ribosomal protein L19
+Macromolecule #45: 50S ribosomal protein L20
+Macromolecule #46: Ribosomal protein L21
+Macromolecule #47: 50S ribosomal protein L22
+Macromolecule #48: 50S ribosomal protein L23
+Macromolecule #49: 50S ribosomal protein L24
+Macromolecule #50: 50S ribosomal protein L25
+Macromolecule #51: 50S ribosomal protein L27
+Macromolecule #52: 50S ribosomal protein L28
+Macromolecule #53: 50S ribosomal protein L29
+Macromolecule #54: 50S ribosomal protein L30
+Macromolecule #55: 50S ribosomal protein L32
+Macromolecule #6: 16S rRNA
+Macromolecule #27: mRNA
+Macromolecule #28: A-site oABZ-tRNAPhe
+Macromolecule #29: P-site fMet-tRNAfMet
+Macromolecule #30: 23S rRNA
+Macromolecule #31: 5S rRNA
+Macromolecule #56: ZINC ION
+Macromolecule #57: PAROMOMYCIN
+Macromolecule #58: MAGNESIUM ION
+Macromolecule #59: 2-AMINO-BENZAMIDE
+Macromolecule #60: 3'-amino-3'-deoxyadenosine 5'-(dihydrogen phosphate)
+Macromolecule #61: N-FORMYLMETHIONINE
+Macromolecule #62: POTASSIUM ION
+Macromolecule #63: SPERMIDINE
+Macromolecule #64: water
-Experimental details
-Structure determination
Method | cryo EM |
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Processing | single particle reconstruction |
Aggregation state | particle |
-Sample preparation
Buffer | pH: 7.5 |
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Grid | Model: Quantifoil R1.2/1.3 / Material: GOLD / Mesh: 300 / Support film - Material: CARBON / Support film - topology: CONTINUOUS / Support film - Film thickness: 0.2 / Pretreatment - Type: GLOW DISCHARGE |
Vitrification | Cryogen name: ETHANE / 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: GATAN K3 BIOQUANTUM (6k x 4k) / Average electron dose: 40.0 e/Å2 |
Electron beam | Acceleration voltage: 300 kV / Electron source: FIELD EMISSION GUN |
Electron optics | Illumination mode: OTHER / Imaging mode: BRIGHT FIELD / Nominal defocus max: 1.5 µm / Nominal defocus min: 0.5 µm |
Experimental equipment | Model: Titan Krios / Image courtesy: FEI Company |