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Yorodumi- PDB-7f0d: Cryo-EM structure of Mycobacterium tuberculosis 50S ribosome subu... -
+Open data
-Basic information
Entry | Database: PDB / ID: 7f0d | ||||||||||||
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Title | Cryo-EM structure of Mycobacterium tuberculosis 50S ribosome subunit bound with clarithromycin | ||||||||||||
Components |
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Keywords | RIBOSOME / Mycobacterium tuberculosis / 50S ribosomal subunit / gate site A2062 / clarithromycin / dynamic interaction / Cryo-EM / drug resistance | ||||||||||||
Function / homology | Function and homology information large ribosomal subunit / 5S rRNA binding / transferase activity / ribosomal large subunit assembly / cytosolic large ribosomal subunit / tRNA binding / rRNA binding / ribosome / structural constituent of ribosome / translation ...large ribosomal subunit / 5S rRNA binding / transferase activity / ribosomal large subunit assembly / cytosolic large ribosomal subunit / tRNA binding / rRNA binding / ribosome / structural constituent of ribosome / translation / ribonucleoprotein complex / metal ion binding / plasma membrane / cytoplasm Similarity search - Function | ||||||||||||
Biological species | Mycobacterium tuberculosis H37Ra (bacteria) | ||||||||||||
Method | ELECTRON MICROSCOPY / single particle reconstruction / cryo EM / Resolution: 3.3 Å | ||||||||||||
Authors | Zhang, W. / Sun, Y. / Gao, N. / Li, Z. | ||||||||||||
Funding support | China, 3items
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Citation | Journal: Emerg Microbes Infect / Year: 2022 Title: Cryo-EM structure of 50S ribosomal subunit bound with clarithromycin reveals dynamic and specific interactions with macrolides. Authors: Wen Zhang / ZhiFei Li / Yufan Sun / Peng Cui / Jianhua Liang / Qinghe Xing / Jing Wu / Yanhui Xu / Wenhong Zhang / Ying Zhang / Lin He / Ning Gao / Abstract: Tuberculosis (TB) is the leading infectious disease caused by (). Clarithromycin (CTY), an analog of erythromycin (ERY), is more potent against multidrug-resistance (MDR) TB. ERY and CTY were ...Tuberculosis (TB) is the leading infectious disease caused by (). Clarithromycin (CTY), an analog of erythromycin (ERY), is more potent against multidrug-resistance (MDR) TB. ERY and CTY were previously reported to bind to the nascent polypeptide exit tunnel (NPET) near peptidyl transferase center (PTC), but the only available CTY structure in complex with () ribosome could be misinterpreted due to resolution limitation. To date, the mechanism of specificity and efficacy of CTY for remains elusive since the ribosome-CTY complex structure is still unknown. Here, we employed new sample preparation methods and solved the ribosome-CTY complex structure at 3.3Å with cryo-EM technique, where the crucial gate site A2062 ( numbering) is located at the CTY binding site within NPET. Two alternative conformations of A2062, a novel -conformation as well as a swayed conformation bound with water molecule at interface, may play a role in coordinating the binding of specific drug molecules. The previously overlooked C-H hydrogen bond (H-bond) and π interaction may collectively contribute to the enhanced binding affinity. Together, our structure data provide a structural basis for the dynamic binding as well as the specificity of CTY and explain of how a single methyl group in CTY improves its potency, which provides new evidence to reveal previously unclear mechanism of translational modulation for future drug design and anti-TB therapy. Furthermore, our sample preparation method may facilitate drug discovery based on the complexes with low water solubility drugs by cryo-EM technique. | ||||||||||||
History |
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-Structure visualization
Structure viewer | Molecule: MolmilJmol/JSmol |
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-Downloads & links
-Download
PDBx/mmCIF format | 7f0d.cif.gz | 2 MB | Display | PDBx/mmCIF format |
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PDB format | pdb7f0d.ent.gz | 1.6 MB | Display | PDB format |
PDBx/mmJSON format | 7f0d.json.gz | Tree view | PDBx/mmJSON format | |
Others | Other downloads |
-Validation report
Summary document | 7f0d_validation.pdf.gz | 1.5 MB | Display | wwPDB validaton report |
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Full document | 7f0d_full_validation.pdf.gz | 1.8 MB | Display | |
Data in XML | 7f0d_validation.xml.gz | 175.3 KB | Display | |
Data in CIF | 7f0d_validation.cif.gz | 284.8 KB | Display | |
Arichive directory | https://data.pdbj.org/pub/pdb/validation_reports/f0/7f0d ftp://data.pdbj.org/pub/pdb/validation_reports/f0/7f0d | HTTPS FTP |
-Related structure data
Related structure data | 31398MC M: map data used to model this data C: citing same article (ref.) |
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Similar structure data | Similarity search - Function & homologyF&H Search |
-Links
-Assembly
Deposited unit |
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1 |
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-Components
+50S ribosomal protein ... , 29 types, 29 molecules 012346CDEFGHJKLMNOPQRSTUVWXYZ
-RNA chain , 2 types, 2 molecules AB
#7: RNA chain | Mass: 1017792.562 Da / Num. of mol.: 1 Source method: isolated from a genetically manipulated source Source: (gene. exp.) Mycobacterium tuberculosis H37Ra (bacteria) Production host: Mycobacterium tuberculosis (bacteria) / References: GenBank: 1251771536 |
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#8: RNA chain | Mass: 37097.238 Da / Num. of mol.: 1 Source method: isolated from a genetically manipulated source Source: (gene. exp.) Mycobacterium tuberculosis H37Ra (bacteria) Production host: Mycobacterium tuberculosis (bacteria) / References: GenBank: 1251771551 |
-Non-polymers , 3 types, 5 molecules
#32: Chemical | ChemComp-CTY / |
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#33: Chemical | ChemComp-MG / |
#34: Water | ChemComp-HOH / |
-Details
Has ligand of interest | Y |
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Source details | The source provided by authors are used. |
-Experimental details
-Experiment
Experiment | Method: ELECTRON MICROSCOPY |
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EM experiment | Aggregation state: PARTICLE / 3D reconstruction method: single particle reconstruction |
-Sample preparation
Component |
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Source (natural) | Organism: Mycobacterium tuberculosis H37Ra (bacteria) | ||||||||||||||||||
Source (recombinant) | Organism: Mycobacterium tuberculosis (bacteria) | ||||||||||||||||||
Buffer solution | pH: 7.5 | ||||||||||||||||||
Specimen | Embedding applied: NO / Shadowing applied: NO / Staining applied: NO / Vitrification applied: YES | ||||||||||||||||||
Vitrification | Cryogen name: NITROGEN |
-Electron microscopy imaging
Experimental equipment | Model: Titan Krios / Image courtesy: FEI Company |
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Microscopy | Model: FEI TITAN KRIOS |
Electron gun | Electron source: FIELD EMISSION GUN / Accelerating voltage: 300 kV / Illumination mode: SPOT SCAN |
Electron lens | Mode: BRIGHT FIELD / Nominal defocus max: 2500 nm / Nominal defocus min: 1500 nm |
Image recording | Electron dose: 50 e/Å2 / Film or detector model: GATAN K2 IS (4k x 4k) |
-Processing
Software | Name: PHENIX / Version: 1.16_3549: / Classification: refinement | ||||||||||||||||||||||||
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EM software |
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CTF correction | Type: NONE | ||||||||||||||||||||||||
3D reconstruction | Resolution: 3.3 Å / Resolution method: FSC 0.143 CUT-OFF / Num. of particles: 34912 / Symmetry type: POINT | ||||||||||||||||||||||||
Atomic model building | PDB-ID: 5V7Q Accession code: 5V7Q / Source name: PDB / Type: experimental model | ||||||||||||||||||||||||
Refine LS restraints |
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