[English] 日本語
Yorodumi- EMDB-3525: Cryo-EM structure of the spinach chloroplast ribosome reveals the... -
+Open data
-Basic information
Entry | Database: EMDB / ID: EMD-3525 | |||||||||||||||
---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
Title | Cryo-EM structure of the spinach chloroplast ribosome reveals the location of plastid-specific ribosomal proteins and extensions | |||||||||||||||
Map data | Reconstruction of the spinach chloroplast 50S ribosomal subunit. | |||||||||||||||
Sample |
| |||||||||||||||
Keywords | Chloroplast / ribosome / ribosomal protein / translation | |||||||||||||||
Function / homology | Function and homology information plastid translation / mitochondrial large ribosomal subunit / mitochondrial translation / chloroplast stroma / chloroplast / DNA-templated transcription termination / large ribosomal subunit / 5S rRNA binding / large ribosomal subunit rRNA binding / transferase activity ...plastid translation / mitochondrial large ribosomal subunit / mitochondrial translation / chloroplast stroma / chloroplast / DNA-templated transcription termination / large ribosomal subunit / 5S rRNA binding / large ribosomal subunit rRNA binding / transferase activity / ribosomal large subunit assembly / cytoplasmic translation / cytosolic large ribosomal subunit / negative regulation of translation / rRNA binding / ribosome / structural constituent of ribosome / ribonucleoprotein complex / translation / mRNA binding / mitochondrion / RNA binding Similarity search - Function | |||||||||||||||
Biological species | Spinacia oleracea (spinach) | |||||||||||||||
Method | single particle reconstruction / cryo EM / Resolution: 3.6 Å | |||||||||||||||
Authors | Graf M / Arenz S | |||||||||||||||
Funding support | Germany, 4 items
| |||||||||||||||
Citation | Journal: Nucleic Acids Res / Year: 2017 Title: Cryo-EM structure of the spinach chloroplast ribosome reveals the location of plastid-specific ribosomal proteins and extensions. Authors: Michael Graf / Stefan Arenz / Paul Huter / Alexandra Dönhöfer / Jirí Novácek / Daniel N Wilson / Abstract: Ribosomes are the protein synthesizing machines of the cell. Recent advances in cryo-EM have led to the determination of structures from a variety of species, including bacterial 70S and eukaryotic ...Ribosomes are the protein synthesizing machines of the cell. Recent advances in cryo-EM have led to the determination of structures from a variety of species, including bacterial 70S and eukaryotic 80S ribosomes as well as mitoribosomes from eukaryotic mitochondria, however, to date high resolution structures of plastid 70S ribosomes have been lacking. Here we present a cryo-EM structure of the spinach chloroplast 70S ribosome, with an average resolution of 5.4 Å for the small 30S subunit and 3.6 Å for the large 50S ribosomal subunit. The structure reveals the location of the plastid-specific ribosomal proteins (RPs) PSRP1, PSRP4, PSRP5 and PSRP6 as well as the numerous plastid-specific extensions of the RPs. We discover many features by which the plastid-specific extensions stabilize the ribosome via establishing additional interactions with surrounding ribosomal RNA and RPs. Moreover, we identify a large conglomerate of plastid-specific protein mass adjacent to the tunnel exit site that could facilitate interaction of the chloroplast ribosome with the thylakoid membrane and the protein-targeting machinery. Comparing the Escherichia coli 70S ribosome with that of the spinach chloroplast ribosome provides detailed insight into the co-evolution of RP and rRNA. | |||||||||||||||
History |
|
-Structure visualization
Movie |
Movie viewer |
---|---|
Structure viewer | EM map: SurfViewMolmilJmol/JSmol |
Supplemental images |
-Downloads & links
-EMDB archive
Map data | emd_3525.map.gz | 6.6 MB | EMDB map data format | |
---|---|---|---|---|
Header (meta data) | emd-3525-v30.xml emd-3525.xml | 46.9 KB 46.9 KB | Display Display | EMDB header |
Images | emd_3525.png | 17.5 KB | ||
Filedesc metadata | emd-3525.cif.gz | 11.5 KB | ||
Archive directory | http://ftp.pdbj.org/pub/emdb/structures/EMD-3525 ftp://ftp.pdbj.org/pub/emdb/structures/EMD-3525 | HTTPS FTP |
-Validation report
Summary document | emd_3525_validation.pdf.gz | 413.8 KB | Display | EMDB validaton report |
---|---|---|---|---|
Full document | emd_3525_full_validation.pdf.gz | 413.4 KB | Display | |
Data in XML | emd_3525_validation.xml.gz | 7.1 KB | Display | |
Data in CIF | emd_3525_validation.cif.gz | 8.2 KB | Display | |
Arichive directory | https://ftp.pdbj.org/pub/emdb/validation_reports/EMD-3525 ftp://ftp.pdbj.org/pub/emdb/validation_reports/EMD-3525 | HTTPS FTP |
-Related structure data
Related structure data | 5mlcMC 3526C M: atomic model generated by this map C: citing same article (ref.) |
---|---|
Similar structure data |
-Links
EMDB pages | EMDB (EBI/PDBe) / EMDataResource |
---|---|
Related items in Molecule of the Month |
-Map
File | Download / File: emd_3525.map.gz / Format: CCP4 / Size: 190.1 MB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES) | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
Annotation | Reconstruction of the spinach chloroplast 50S ribosomal subunit. | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Projections & slices | Image control
Images are generated by Spider. | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Voxel size | X=Y=Z: 1.061 Å | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Density |
| ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Symmetry | Space group: 1 | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Details | EMDB XML:
CCP4 map header:
|
-Supplemental data
-Sample components
+Entire : 50S subunit of the spinach chloroplast ribosome
+Supramolecule #1: 50S subunit of the spinach chloroplast ribosome
+Macromolecule #1: 23S ribosomal RNA, chloroplastic
+Macromolecule #2: 5S ribosomal RNA, chloroplastic
+Macromolecule #3: 4.8S ribosomal RNA, chloroplastic
+Macromolecule #4: 50S ribosomal protein L2, chloroplastic
+Macromolecule #5: 50S ribosomal protein L3, chloroplastic
+Macromolecule #6: 50S ribosomal protein L4, chloroplastic
+Macromolecule #7: 50S ribosomal protein L5, chloroplastic
+Macromolecule #8: 50S ribosomal protein L6, chloroplastic
+Macromolecule #9: 50S ribosomal protein L9, chloroplastic
+Macromolecule #10: 50S ribosomal protein L13, chloroplastic
+Macromolecule #11: 50S ribosomal protein L14, chloroplastic
+Macromolecule #12: 50S ribosomal protein L15, chloroplastic
+Macromolecule #13: 50S ribosomal protein L16, chloroplastic
+Macromolecule #14: 50S ribosomal protein L17, chloroplastic
+Macromolecule #15: 50S ribosomal protein L18, chloroplastic
+Macromolecule #16: 50S ribosomal protein L19, chloroplastic
+Macromolecule #17: 50S ribosomal protein L20, chloroplastic
+Macromolecule #18: 50S ribosomal protein L21, chloroplastic
+Macromolecule #19: 50S ribosomal protein L22, chloroplastic
+Macromolecule #20: 50S ribosomal protein L23, chloroplastic
+Macromolecule #21: 50S ribosomal protein L24, chloroplastic
+Macromolecule #22: 50S ribosomal protein L27, chloroplastic
+Macromolecule #23: 50S ribosomal protein L28, chloroplastic
+Macromolecule #24: 50S ribosomal protein L29, chloroplastic
+Macromolecule #25: 50S ribosomal protein L32, chloroplastic
+Macromolecule #26: 50S ribosomal protein L33, chloroplastic
+Macromolecule #27: 50S ribosomal protein L34, chloroplastic
+Macromolecule #28: 50S ribosomal protein L35, chloroplastic
+Macromolecule #29: 50S ribosomal protein L36, chloroplastic
+Macromolecule #30: Ribosome-recycling factor, chloroplastic
+Macromolecule #31: PSRP5alpha, chloroplastic
+Macromolecule #32: PSRP6, chloroplastic
-Experimental details
-Structure determination
Method | cryo EM |
---|---|
Processing | single particle reconstruction |
Aggregation state | particle |
-Sample preparation
Buffer | pH: 7.4 Component:
Details: Solutions were made fresh and filtered previous to usage. | |||||||||||||||
---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
Grid | Model: Quantifoil R3/3 / Material: COPPER / Support film - Material: CARBON / Support film - topology: HOLEY | |||||||||||||||
Vitrification | Cryogen name: ETHANE / Instrument: FEI VITROBOT MARK IV |
-Electron microscopy
Microscope | FEI TITAN KRIOS |
---|---|
Image recording | Film or detector model: FEI FALCON II (4k x 4k) / Detector mode: INTEGRATING / Digitization - Dimensions - Width: 4096 pixel / Digitization - Dimensions - Height: 4096 pixel / Average electron dose: 2.6 e/Å2 |
Electron beam | Acceleration voltage: 300 kV / Electron source: FIELD EMISSION GUN |
Electron optics | Illumination mode: SPOT SCAN / Imaging mode: BRIGHT FIELD / Cs: 2.7 mm |
Experimental equipment | Model: Titan Krios / Image courtesy: FEI Company |
-Image processing
Startup model | Type of model: PDB ENTRY PDB model - PDB ID: |
---|---|
Final reconstruction | Applied symmetry - Point group: C1 (asymmetric) / Resolution.type: BY AUTHOR / Resolution: 3.6 Å / Resolution method: FSC 0.143 CUT-OFF / Software - Name: FREALIGN (ver. 9.11) / Number images used: 37636 |
Initial angle assignment | Type: PROJECTION MATCHING |
Final angle assignment | Type: PROJECTION MATCHING |
-Atomic model buiding 1
Refinement | Space: REAL / Protocol: RIGID BODY FIT |
---|---|
Output model | PDB-5mlc: |