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Yorodumi- EMDB-10375: Complex IV in the III2-IV2 mitochondrial respiratory supercomplex... -
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Open data
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Basic information
| Entry | Database: EMDB / ID: EMD-10375 | |||||||||
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| Title | Complex IV in the III2-IV2 mitochondrial respiratory supercomplex from S. cerevisiae (CIV(5A)-a) | |||||||||
Map data | The reprocessed CIVa map after focused refinement with a mask including rcf2 | |||||||||
Sample |
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| Biological species | ![]() | |||||||||
| Method | single particle reconstruction / cryo EM / Resolution: 3.09 Å | |||||||||
Authors | Marechal A / Hartley A / Pinotsis N | |||||||||
| Funding support | United Kingdom, 1 items
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Citation | Journal: Proc Natl Acad Sci U S A / Year: 2020Title: Rcf2 revealed in cryo-EM structures of hypoxic isoforms of mature mitochondrial III-IV supercomplexes. Authors: Andrew M Hartley / Brigitte Meunier / Nikos Pinotsis / Amandine Maréchal / ![]() Abstract: The organization of the mitochondrial electron transport chain proteins into supercomplexes (SCs) is now undisputed; however, their assembly process, or the role of differential expression isoforms, ...The organization of the mitochondrial electron transport chain proteins into supercomplexes (SCs) is now undisputed; however, their assembly process, or the role of differential expression isoforms, remain to be determined. In , cytochrome oxidase (CIV) forms SCs of varying stoichiometry with cytochrome (CIII). Recent studies have revealed, in normoxic growth conditions, an interface made exclusively by Cox5A, the only yeast respiratory protein that exists as one of two isoforms depending on oxygen levels. Here we present the cryo-EM structures of the III-IV and III-IV SCs containing the hypoxic isoform Cox5B solved at 3.4 and 2.8 Å, respectively. We show that the change of isoform does not affect SC formation or activity, and that SC stoichiometry is dictated by the level of CIII/CIV biosynthesis. Comparison of the CIV- and CIV-containing SC structures highlighted few differences, found mainly in the region of Cox5. Additional density was revealed in all SCs, independent of the CIV isoform, in a pocket formed by Cox1, Cox3, Cox12, and Cox13, away from the CIII-CIV interface. In the CIV-containing hypoxic SCs, this could be confidently assigned to the hypoxia-induced gene 1 (Hig1) type 2 protein Rcf2. With conserved residues in mammalian Hig1 proteins and Cox3/Cox12/Cox13 orthologs, we propose that Hig1 type 2 proteins are stoichiometric subunits of CIV, at least when within a III-IV SC. | |||||||||
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Structure visualization
| Movie |
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| Structure viewer | EM map: SurfView Molmil Jmol/JSmol |
| Supplemental images |
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Downloads & links
-EMDB archive
| Map data | emd_10375.map.gz | 483.6 MB | EMDB map data format | |
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| Header (meta data) | emd-10375-v30.xml emd-10375.xml | 23 KB 23 KB | Display Display | EMDB header |
| FSC (resolution estimation) | emd_10375_fsc.xml | 17.8 KB | Display | FSC data file |
| Images | emd_10375.png | 110.6 KB | ||
| Archive directory | http://ftp.pdbj.org/pub/emdb/structures/EMD-10375 ftp://ftp.pdbj.org/pub/emdb/structures/EMD-10375 | HTTPS FTP |
-Validation report
| Summary document | emd_10375_validation.pdf.gz | 295.9 KB | Display | EMDB validaton report |
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| Full document | emd_10375_full_validation.pdf.gz | 295.1 KB | Display | |
| Data in XML | emd_10375_validation.xml.gz | 16.4 KB | Display | |
| Arichive directory | https://ftp.pdbj.org/pub/emdb/validation_reports/EMD-10375 ftp://ftp.pdbj.org/pub/emdb/validation_reports/EMD-10375 | HTTPS FTP |
-Related structure data
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Links
| EMDB pages | EMDB (EBI/PDBe) / EMDataResource |
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Map
| File | Download / File: emd_10375.map.gz / Format: CCP4 / Size: 512 MB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES) | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
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| Annotation | The reprocessed CIVa map after focused refinement with a mask including rcf2 | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
| Projections & slices | Image control
Images are generated by Spider. | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
| Voxel size | X=Y=Z: 1.048 Å | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
| Density |
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| Symmetry | Space group: 1 | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
| Details | EMDB XML:
CCP4 map header:
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-Supplemental data
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Sample components
+Entire : Complex IV in the III2-IV2 mitochondrial respiratory supercomplex...
+Supramolecule #1: Complex IV in the III2-IV2 mitochondrial respiratory supercomplex...
+Supramolecule #2: Complex IV in the III2-IV2 mitochondrial respiratory supercomplex...
+Supramolecule #3: Complex IV in the III2-IV2 mitochondrial respiratory supercomplex...
+Macromolecule #1: CYTOCHROME C OXIDASE SUBUNIT 1; SYNONYM: CYTOCHROME C OXIDASE POL...
+Macromolecule #2: CYTOCHROME C OXIDASE SUBUNIT 2; SYNONYM: CYTOCHROME C OXIDASE POL...
+Macromolecule #3: CYTOCHROME C OXIDASE SUBUNIT 3; SYNONYM: CYTOCHROME C OXIDASE POL...
+Macromolecule #4: CYTOCHROME C OXIDASE SUBUNIT 4, MITOCHONDRIAL; SYNONYM: CYTOCHROM...
+Macromolecule #5: CYTOCHROME C OXIDASE POLYPEPTIDE 5A, MITOCHONDRIAL; SYNONYM: CYTO...
+Macromolecule #6: CYTOCHROME C OXIDASE SUBUNIT 6, MITOCHONDRIAL; SYNONYM: CYTOCHROM...
+Macromolecule #7: CYTOCHROME C OXIDASE SUBUNIT 7; SYNONYM: CYTOCHROME C OXIDASE POL...
+Macromolecule #8: CYTOCHROME C OXIDASE POLYPEPTIDE VIII, MITOCHONDRIAL; SYNONYM: COX8
+Macromolecule #9: CYTOCHROME C OXIDASE SUBUNIT 7A; SYNONYM: CYTOCHROME C OXIDASE PO...
+Macromolecule #10: CYTOCHROME C OXIDASE SUBUNIT 6B; SYNONYM: CYTOCHROME C OXIDASE PO...
+Macromolecule #11: CYTOCHROME C OXIDASE SUBUNIT 6A, MITOCHONDRIAL; CYTOCHROME C OXID...
+Macromolecule #12: COX26; SYNONYM: Uncharacterized protein YDR119W-A
+Macromolecule #13: RCF2; SYNONYM: Respiratory supercomplex factor 2
-Experimental details
-Structure determination
| Method | cryo EM |
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Processing | single particle reconstruction |
| Aggregation state | particle |
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Sample preparation
| Buffer | pH: 7.2 |
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| Vitrification | Cryogen name: ETHANE / Instrument: FEI VITROBOT MARK IV Details: 3 uL of sample applied to negatively glow discharged grid, blot force -10; blotting time 8.5 sec.. |
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Electron microscopy
| Microscope | FEI TITAN KRIOS |
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| Image recording | Film or detector model: GATAN K2 SUMMIT (4k x 4k) / Number real images: 2740 / Average electron dose: 52.64 e/Å2 |
| Electron beam | Acceleration voltage: 300 kV / Electron source: FIELD EMISSION GUN |
| Electron optics | Illumination mode: FLOOD BEAM / Imaging mode: BRIGHT FIELD |
| Experimental equipment | ![]() Model: Titan Krios / Image courtesy: FEI Company |
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Authors
United Kingdom, 1 items
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