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Yorodumi- EMDB-11969: A 3.7-angstrom structure of Yarrowia lipolytica complex I with an... -
+Open data
-Basic information
Entry | Database: EMDB / ID: EMD-11969 | |||||||||
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Title | A 3.7-angstrom structure of Yarrowia lipolytica complex I with an R121M mutation in NUCM. | |||||||||
Map data | Final map globally sharpened with RELION postprocess | |||||||||
Sample |
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Keywords | NADH:Ubiquinone Oxidoreductase / complex I / MEMBRANE PROTEIN | |||||||||
Function / homology | Function and homology information NADH dehydrogenase / oxidoreductase activity, acting on NAD(P)H / NADH:ubiquinone reductase (H+-translocating) / respiratory chain complex I / : / mitochondrial respiratory chain complex I assembly / mitochondrial electron transport, NADH to ubiquinone / NADH dehydrogenase (ubiquinone) activity / ATP synthesis coupled electron transport / quinone binding ...NADH dehydrogenase / oxidoreductase activity, acting on NAD(P)H / NADH:ubiquinone reductase (H+-translocating) / respiratory chain complex I / : / mitochondrial respiratory chain complex I assembly / mitochondrial electron transport, NADH to ubiquinone / NADH dehydrogenase (ubiquinone) activity / ATP synthesis coupled electron transport / quinone binding / : / respiratory electron transport chain / mitochondrial membrane / electron transport chain / mitochondrial intermembrane space / fatty acid biosynthetic process / 2 iron, 2 sulfur cluster binding / NAD binding / FMN binding / 4 iron, 4 sulfur cluster binding / mitochondrial inner membrane / oxidoreductase activity / mitochondrion / membrane / metal ion binding Similarity search - Function | |||||||||
Biological species | Yarrowia lipolytica (yeast) | |||||||||
Method | single particle reconstruction / cryo EM / Resolution: 3.7 Å | |||||||||
Authors | Hirst J / Grba D | |||||||||
Funding support | United Kingdom, 2 items
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Citation | Journal: J Biol Chem / Year: 2021 Title: A conserved arginine residue is critical for stabilizing the N2 FeS cluster in mitochondrial complex I. Authors: Mikhail A Hameedi / Daniel N Grba / Katherine H Richardson / Andrew J Y Jones / Wei Song / Maxie M Roessler / John J Wright / Judy Hirst / Abstract: Respiratory complex I (NADH:ubiquinone oxidoreductase), the first enzyme of the electron-transport chain, captures the free energy released by NADH oxidation and ubiquinone reduction to translocate ...Respiratory complex I (NADH:ubiquinone oxidoreductase), the first enzyme of the electron-transport chain, captures the free energy released by NADH oxidation and ubiquinone reduction to translocate protons across an energy-transducing membrane and drive ATP synthesis during oxidative phosphorylation. The cofactor that transfers the electrons directly to ubiquinone is an iron-sulfur cluster (N2) located in the NDUFS2/NUCM subunit. A nearby arginine residue (R121), which forms part of the second coordination sphere of the N2 cluster, is known to be posttranslationally dimethylated but its functional and structural significance are not known. Here, we show that mutations of this arginine residue (R121M/K) abolish the quinone-reductase activity, concomitant with disappearance of the N2 signature from the electron paramagnetic resonance (EPR) spectrum. Analysis of the cryo-EM structure of NDUFS2-R121M complex I at 3.7 Å resolution identified the absence of the cubane N2 cluster as the cause of the dysfunction, within an otherwise intact enzyme. The mutation further induced localized disorder in nearby elements of the quinone-binding site, consistent with the close connections between the cluster and substrate-binding regions. Our results demonstrate that R121 is required for the formation and/or stability of the N2 cluster and highlight the importance of structural analyses for mechanistic interpretation of biochemical and spectroscopic data on complex I variants. | |||||||||
History |
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-Structure visualization
Movie |
Movie viewer |
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Structure viewer | EM map: SurfViewMolmilJmol/JSmol |
Supplemental images |
-Downloads & links
-EMDB archive
Map data | emd_11969.map.gz | 323.7 MB | EMDB map data format | |
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Header (meta data) | emd-11969-v30.xml emd-11969.xml | 74.5 KB 74.5 KB | Display Display | EMDB header |
FSC (resolution estimation) | emd_11969_fsc.xml | 16 KB | Display | FSC data file |
Images | emd_11969.png | 49.3 KB | ||
Masks | emd_11969_msk_1.map | 347.6 MB | Mask map | |
Filedesc metadata | emd-11969.cif.gz | 15 KB | ||
Others | emd_11969_half_map_1.map.gz emd_11969_half_map_2.map.gz | 277.6 MB 277.8 MB | ||
Archive directory | http://ftp.pdbj.org/pub/emdb/structures/EMD-11969 ftp://ftp.pdbj.org/pub/emdb/structures/EMD-11969 | HTTPS FTP |
-Validation report
Summary document | emd_11969_validation.pdf.gz | 1.2 MB | Display | EMDB validaton report |
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Full document | emd_11969_full_validation.pdf.gz | 1.2 MB | Display | |
Data in XML | emd_11969_validation.xml.gz | 23 KB | Display | |
Data in CIF | emd_11969_validation.cif.gz | 31 KB | Display | |
Arichive directory | https://ftp.pdbj.org/pub/emdb/validation_reports/EMD-11969 ftp://ftp.pdbj.org/pub/emdb/validation_reports/EMD-11969 | HTTPS FTP |
-Related structure data
Related structure data | 7b0nMC M: atomic model generated by this map C: citing same article (ref.) |
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Similar structure data |
-Links
EMDB pages | EMDB (EBI/PDBe) / EMDataResource |
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Related items in Molecule of the Month |
-Map
File | Download / File: emd_11969.map.gz / Format: CCP4 / Size: 347.6 MB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES) | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
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Annotation | Final map globally sharpened with RELION postprocess | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Projections & slices | Image control
Images are generated by Spider. | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Voxel size | X=Y=Z: 1.05 Å | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Density |
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Symmetry | Space group: 1 | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Details | EMDB XML:
CCP4 map header:
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-Supplemental data
-Mask #1
File | emd_11969_msk_1.map | ||||||||||||
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Projections & Slices |
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Density Histograms |
-Half map: Half map 1
File | emd_11969_half_map_1.map | ||||||||||||
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Annotation | Half map 1 | ||||||||||||
Projections & Slices |
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Density Histograms |
-Half map: Half map 2
File | emd_11969_half_map_2.map | ||||||||||||
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Annotation | Half map 2 | ||||||||||||
Projections & Slices |
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Density Histograms |
-Sample components
+Entire : Yarrowia lipolytica mitochondrial complex I (NADH:Ubiquinone oxid...
+Supramolecule #1: Yarrowia lipolytica mitochondrial complex I (NADH:Ubiquinone oxid...
+Macromolecule #1: NADH-ubiquinone oxidoreductase chain 3
+Macromolecule #2: Subunit NUKM of NADH:Ubiquinone Oxidoreductase (Complex I)
+Macromolecule #3: NUGM protein
+Macromolecule #4: NUCM protein
+Macromolecule #5: Subunit NUHM of NADH:Ubiquinone Oxidoreductase (Complex I)
+Macromolecule #6: Subunit NUBM of NADH:Ubiquinone Oxidoreductase (Complex I)
+Macromolecule #7: Subunit NUAM of NADH:Ubiquinone Oxidoreductase (Complex I)
+Macromolecule #8: NADH-ubiquinone oxidoreductase chain 1
+Macromolecule #9: Subunit NUIM of protein NADH:Ubiquinone Oxidoreductase (Complex I)
+Macromolecule #10: NADH-ubiquinone oxidoreductase chain 6
+Macromolecule #11: NADH-ubiquinone oxidoreductase chain 4L
+Macromolecule #12: NADH-ubiquinone oxidoreductase chain 5
+Macromolecule #13: NADH-ubiquinone oxidoreductase chain 4
+Macromolecule #14: NADH dehydrogenase subunit 2
+Macromolecule #15: Subunit NUXM of NADH:Ubiquinone Oxidoreductase (Complex I)
+Macromolecule #16: Epimerase domain-containing protein
+Macromolecule #17: Subunit NUYM of NADH:Ubiquinone Oxidoreductase (Complex I)
+Macromolecule #18: zf-CHCC domain-containing protein
+Macromolecule #19: Subunit NI8M of NADH:Ubiquinone Oxidoreductase (Complex I)
+Macromolecule #20: Acyl carrier protein
+Macromolecule #21: Acyl carrier protein
+Macromolecule #22: Subunit NUFM of NADH:Ubiquinone Oxidoreductase (Complex I)
+Macromolecule #23: Subunit NB4M of protein NADH:Ubiquinone Oxidoreductase (Complex I)
+Macromolecule #24: Subunit NUPM of NADH:Ubiquinone Oxidoreductase (Complex I)
+Macromolecule #25: Complex I-B14.7
+Macromolecule #26: GRIM-19
+Macromolecule #27: NADH dehydrogenase [ubiquinone] 1 alpha subcomplex subunit 1
+Macromolecule #28: subunit NI9M of protein NADH:Ubiquinone Oxidoreductase (Complex I)
+Macromolecule #29: Subunit NUZM of NADH:Ubiquinone Oxidoreductase (Complex I)
+Macromolecule #30: subunit NEBM of protein NADH:Ubiquinone Oxidoreductase (Complex I...
+Macromolecule #31: Subunit NIPM of NADH:Ubiquinone Oxidoreductase (Complex I)
+Macromolecule #32: Subunit N7BM of NADH:Ubiquinone Oxidoreductase (Complex I)
+Macromolecule #33: Subunit NESM of NADH:Ubiquinone Oxidoreductase (Complex I)
+Macromolecule #34: subunit NUNM of protein NADH:Ubiquinone Oxidoreductase (Complex I)
+Macromolecule #35: Subunit NUUM of NADH:Ubiquinone Oxidoreductase (Complex I)
+Macromolecule #36: Subunit NUVM of NADH:Ubiquinone Oxidoreductase (Complex I)
+Macromolecule #37: Subunit NB2M of NADH:Ubiquinone Oxidoreductase (Complex I)
+Macromolecule #38: Subunit NIAM of NADH:Ubiquinone Oxidoreductase (Complex I)
+Macromolecule #39: Subunit NB5M of NADH:Ubiquinone Oxidoreductase (Complex I)
+Macromolecule #40: Subunit NI2M of NADH:Ubiquinone Oxidoreductase (Complex I)
+Macromolecule #41: Subunit NB8M of NADH:Ubiquinone Oxidoreductase (Complex I)
+Macromolecule #42: Subunit NIDM of NADH:Ubiquinone Oxidoreductase (Complex I)
+Macromolecule #43: DIUNDECYL PHOSPHATIDYL CHOLINE
+Macromolecule #44: FE2/S2 (INORGANIC) CLUSTER
+Macromolecule #45: FLAVIN MONONUCLEOTIDE
+Macromolecule #46: IRON/SULFUR CLUSTER
+Macromolecule #47: 1,2-Distearoyl-sn-glycerophosphoethanolamine
+Macromolecule #48: CARDIOLIPIN
+Macromolecule #49: NADPH DIHYDRO-NICOTINAMIDE-ADENINE-DINUCLEOTIDE PHOSPHATE
+Macromolecule #50: ZINC ION
+Macromolecule #51: ~{S}-[2-[3-[[(2~{R})-3,3-dimethyl-2-oxidanyl-4-phosphonooxy-butan...
+Macromolecule #52: DODECYL-BETA-D-MALTOSIDE
-Experimental details
-Structure determination
Method | cryo EM |
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Processing | single particle reconstruction |
Aggregation state | particle |
-Sample preparation
Buffer | pH: 7.45 |
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Grid | Model: UltrAuFoil / Material: GOLD / Support film - Material: GOLD / Pretreatment - Type: PLASMA CLEANING / Pretreatment - Time: 90 sec. / Details: PEG-thiol treated |
Vitrification | Cryogen name: ETHANE |
-Electron microscopy
Microscope | FEI TITAN KRIOS |
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Image recording | Film or detector model: GATAN K2 SUMMIT (4k x 4k) / Detector mode: COUNTING / Number grids imaged: 1 / Number real images: 2241 / Average electron dose: 48.0 e/Å2 |
Electron beam | Acceleration voltage: 300 kV / Electron source: FIELD EMISSION GUN |
Electron optics | C2 aperture diameter: 50.0 µm / Calibrated defocus max: -2.7 µm / Calibrated defocus min: -1.5 µm / Illumination mode: FLOOD BEAM / Imaging mode: BRIGHT FIELD / Cs: 2.7 mm / Nominal magnification: 130000 |
Experimental equipment | Model: Titan Krios / Image courtesy: FEI Company |