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- PDB-7b0n: A 3.7-angstrom structure of Yarrowia lipolytica complex I with an... -
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Basic information
Entry | Database: PDB / ID: 7b0n | ||||||||||||
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Title | A 3.7-angstrom structure of Yarrowia lipolytica complex I with an R121M mutation in NUCM. | ||||||||||||
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![]() | MEMBRANE PROTEIN / NADH:Ubiquinone Oxidoreductase / complex I | ||||||||||||
Function / homology | ![]() NADH dehydrogenase / ubiquinone-6 biosynthetic process / respiratory chain complex I / oxidoreductase activity, acting on NAD(P)H / NADH dehydrogenase activity / NADH:ubiquinone reductase (H+-translocating) / : / mitochondrial electron transport, NADH to ubiquinone / mitochondrial respiratory chain complex I assembly / ubiquinone binding ...NADH dehydrogenase / ubiquinone-6 biosynthetic process / respiratory chain complex I / oxidoreductase activity, acting on NAD(P)H / NADH dehydrogenase activity / NADH:ubiquinone reductase (H+-translocating) / : / mitochondrial electron transport, NADH to ubiquinone / mitochondrial respiratory chain complex I assembly / ubiquinone binding / acyl binding / NADH dehydrogenase (ubiquinone) activity / acyl carrier activity / quinone binding / : / electron transport coupled proton transport / ATP synthesis coupled electron transport / aerobic respiration / respiratory electron transport chain / mitochondrial membrane / electron transport chain / mitochondrial intermembrane space / 2 iron, 2 sulfur cluster binding / NAD binding / FMN binding / 4 iron, 4 sulfur cluster binding / response to oxidative stress / oxidoreductase activity / protein-containing complex binding / mitochondrion / membrane / metal ion binding Similarity search - Function | ||||||||||||
Biological species | ![]() | ||||||||||||
Method | ELECTRON MICROSCOPY / single particle reconstruction / cryo EM / Resolution: 3.7 Å | ||||||||||||
![]() | Hirst, J. / Grba, D. | ||||||||||||
Funding support | ![]()
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![]() | ![]() 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
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Structure viewer | Molecule: ![]() ![]() |
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PDBx/mmCIF format | ![]() | 2.6 MB | Display | ![]() |
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PDB format | ![]() | Display | ![]() | |
PDBx/mmJSON format | ![]() | Tree view | ![]() | |
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-Validation report
Arichive directory | ![]() ![]() | HTTPS FTP |
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-Related structure data
Related structure data | ![]() 11969MC M: map data used to model this data C: citing same article ( |
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Similar structure data |
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Links
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Assembly
Deposited unit | ![]()
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Components
-NADH-ubiquinone oxidoreductase chain ... , 6 types, 6 molecules AHJKLM
#1: Protein | Mass: 14506.339 Da / Num. of mol.: 1 Source method: isolated from a genetically manipulated source Source: (gene. exp.) ![]() ![]() References: UniProt: S5TMS4, NADH:ubiquinone reductase (H+-translocating) |
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#8: Protein | Mass: 38389.277 Da / Num. of mol.: 1 Source method: isolated from a genetically manipulated source Details: ligands (PLC)(3PE)(3PE) are not in the polypeptide sequence (FME) is at the start of the polypeptide sequence Source: (gene. exp.) ![]() ![]() References: UniProt: S5U3V2, NADH:ubiquinone reductase (H+-translocating) |
#10: Protein | Mass: 20793.111 Da / Num. of mol.: 1 Source method: isolated from a genetically manipulated source Details: (FME) is in the polypeptide sequence / Source: (gene. exp.) ![]() ![]() References: UniProt: S5U3X7, NADH:ubiquinone reductase (H+-translocating) |
#11: Protein | Mass: 9843.860 Da / Num. of mol.: 1 Source method: isolated from a genetically manipulated source Details: (FME) is in the polypeptide sequence / Source: (gene. exp.) ![]() ![]() References: UniProt: S5U4U1, NADH:ubiquinone reductase (H+-translocating) |
#12: Protein | Mass: 73768.703 Da / Num. of mol.: 1 Source method: isolated from a genetically manipulated source Details: ligands (PLC)(3PE)(3PE)(3PE)(3PE) are not in the polypeptide sequence (FME) is in the polypeptide sequence Source: (gene. exp.) ![]() ![]() References: UniProt: S5TF58, NADH:ubiquinone reductase (H+-translocating) |
#13: Protein | Mass: 54534.652 Da / Num. of mol.: 1 Source method: isolated from a genetically manipulated source Details: ligands (CDL)(PLC)(PLC)(3PE)(LMT)(3PE) are not in the polypeptide sequence (FME) is in the polypeptide sequence Source: (gene. exp.) ![]() ![]() References: UniProt: S5TMP9, NADH:ubiquinone reductase (H+-translocating) |
+Protein , 32 types, 32 molecules BCDEFGIOPQRSVWXYZbcdefghijklmnop
-NADH dehydrogenase ... , 2 types, 2 molecules Na
#14: Protein | Mass: 53381.367 Da / Num. of mol.: 1 Source method: isolated from a genetically manipulated source Details: ligands (PLC)(PLC)(3PE) are not in the polypeptide sequence (FME) is in the polypeptide sequence Source: (gene. exp.) ![]() ![]() References: UniProt: S5U4R9, NADH:ubiquinone reductase (H+-translocating) |
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#27: Protein | Mass: 9675.043 Da / Num. of mol.: 1 Source method: isolated from a genetically manipulated source Details: Ligand (CDL) is not in the polypeptide chain / Source: (gene. exp.) ![]() ![]() |
-Acyl carrier ... , 2 types, 2 molecules TU
#20: Protein | Mass: 8821.807 Da / Num. of mol.: 1 Source method: isolated from a genetically manipulated source Details: (EHZ) is not in the polypeptide chain / Source: (gene. exp.) ![]() ![]() |
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#21: Protein | Mass: 9533.592 Da / Num. of mol.: 1 Source method: isolated from a genetically manipulated source Details: (EHZ) is not in the polypeptide chain / Source: (gene. exp.) ![]() ![]() |
-Sugars , 1 types, 2 molecules ![](data/chem/img/LMT.gif)
#52: Sugar |
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-Non-polymers , 9 types, 36 molecules ![](data/chem/img/PLC.gif)
![](data/chem/img/FES.gif)
![](data/chem/img/FMN.gif)
![](data/chem/img/SF4.gif)
![](data/chem/img/3PE.gif)
![](data/chem/img/CDL.gif)
![](data/chem/img/NDP.gif)
![](data/chem/img/ZN.gif)
![](data/chem/img/EHZ.gif)
![](data/chem/img/FES.gif)
![](data/chem/img/FMN.gif)
![](data/chem/img/SF4.gif)
![](data/chem/img/3PE.gif)
![](data/chem/img/CDL.gif)
![](data/chem/img/NDP.gif)
![](data/chem/img/ZN.gif)
![](data/chem/img/EHZ.gif)
#43: Chemical | ChemComp-PLC / #44: Chemical | #45: Chemical | ChemComp-FMN / | #46: Chemical | ChemComp-SF4 / #47: Chemical | ChemComp-3PE / #48: Chemical | ChemComp-CDL / #49: Chemical | ChemComp-NDP / | #50: Chemical | ChemComp-ZN / | #51: Chemical | |
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-Details
Has ligand of interest | N |
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-Experimental details
-Experiment
Experiment | Method: ELECTRON MICROSCOPY |
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EM experiment | Aggregation state: PARTICLE / 3D reconstruction method: single particle reconstruction |
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Sample preparation
Component | Name: Yarrowia lipolytica mitochondrial complex I (NADH:Ubiquinone oxidoreductase) with NUCM R121M mutation Type: COMPLEX Entity ID: #1, #10-#11, #15-#19, #2, #20-#29, #3, #30-#39, #4, #40-#42, #5-#9 Source: RECOMBINANT |
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Source (natural) | Organism: ![]() |
Source (recombinant) | Organism: ![]() |
Buffer solution | pH: 7.45 |
Specimen | Embedding applied: NO / Shadowing applied: NO / Staining applied: NO / Vitrification applied: YES |
Specimen support | Details: PEG-thiol treated / Grid material: GOLD / Grid type: UltrAuFoil |
Vitrification | Cryogen name: ETHANE |
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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: ![]() |
Electron lens | Mode: BRIGHT FIELD / Nominal magnification: 130000 X / Calibrated defocus min: -1500 nm / Calibrated defocus max: -2700 nm / Cs: 2.7 mm / C2 aperture diameter: 50 µm |
Image recording | Electron dose: 48 e/Å2 / Detector mode: COUNTING / Film or detector model: GATAN K2 SUMMIT (4k x 4k) / Num. of grids imaged: 1 / Num. of real images: 2241 |
Image scans | Movie frames/image: 40 |
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Processing
EM software |
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CTF correction | Type: PHASE FLIPPING AND AMPLITUDE CORRECTION | ||||||||||||||||||||||||||||||||||||||||||||
Particle selection | Num. of particles selected: 110787 / Details: Post manual curation of Relion auto-pick | ||||||||||||||||||||||||||||||||||||||||||||
3D reconstruction | Resolution: 3.7 Å / Resolution method: FSC 0.143 CUT-OFF / Num. of particles: 21013 / Num. of class averages: 1 / Symmetry type: POINT | ||||||||||||||||||||||||||||||||||||||||||||
Atomic model building | PDB-ID: 6YJ4 |