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- PDB-9ihr: Closed state with NUQM and with flavoprotein (classification stat... -
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Open data
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Basic information
Entry | Database: PDB / ID: 9ihr | ||||||
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Title | Closed state with NUQM and with flavoprotein (classification state 1) of Pichia pastoris mitochondrial complex I in cMSP26 nanodiscs | ||||||
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![]() | MEMBRANE PROTEIN / NADH:ubiquinone oxidoreductase nanodisc membrane protein complex | ||||||
Function / homology | ![]() carbon-nitrogen ligase activity, with glutamine as amido-N-donor / NADH:ubiquinone reductase (H+-translocating) / NADH dehydrogenase complex / oxidoreductase activity, acting on NAD(P)H / NADH dehydrogenase activity / acyl binding / ubiquinone binding / electron transport coupled proton transport / acyl carrier activity / NADH:ubiquinone reductase (H+-translocating) ...carbon-nitrogen ligase activity, with glutamine as amido-N-donor / NADH:ubiquinone reductase (H+-translocating) / NADH dehydrogenase complex / oxidoreductase activity, acting on NAD(P)H / NADH dehydrogenase activity / acyl binding / ubiquinone binding / electron transport coupled proton transport / acyl carrier activity / NADH:ubiquinone reductase (H+-translocating) / mitochondrial respiratory chain complex I assembly / mitochondrial electron transport, NADH to ubiquinone / respiratory chain complex I / NADH dehydrogenase (ubiquinone) activity / quinone binding / ATP synthesis coupled electron transport / aerobic respiration / respiratory electron transport chain / electron transport chain / mitochondrial membrane / 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 / mitochondrial inner membrane / protein-containing complex binding / mitochondrion / metal ion binding / membrane Similarity search - Function | ||||||
Biological species | ![]() | ||||||
Method | ELECTRON MICROSCOPY / single particle reconstruction / cryo EM / Resolution: 2.8 Å | ||||||
![]() | Grba, D.N. / Hirst, J. | ||||||
Funding support | ![]()
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![]() | ![]() Title: Global conformations of complex I are distinguished by the binding of a unique interdomain bridging subunit. Authors: Chris Seunggyu Lee / Daniel N Grba / John J Wright / Bozhidar S Ivanov / Judy Hirst / ![]() Abstract: Complex I (CI; NADH ubiquinone oxidoreductase) is central to energy generation and metabolic homeostasis in mammalian cells but contributes to adverse outcome pathways under challenging conditions. ...Complex I (CI; NADH ubiquinone oxidoreductase) is central to energy generation and metabolic homeostasis in mammalian cells but contributes to adverse outcome pathways under challenging conditions. During ischemia, mammalian CI transitions from a turnover-ready, structurally "closed" state toward a dormant "open" state that prevents it from functioning in reverse during reperfusion to produce reactive oxygen species. Unfortunately, simpler, genetically tractable CI models do not recapitulate the same regulatory behavior, compromising mechanistic studies. Here, we report the structure of isolated CI from the yeast (-CI) and identify distinct closed and open states that resemble those of mammalian CI. Notably, a hitherto-unknown protein (NUQM) completes an interdomain bridge in only the closed state, implying that NUQM stabilizes it by restricting the conformational changes of opening. The direct correlation of NUQM binding with closed/open status in -CI provides opportunities for investigating regulatory mechanisms relevant to reversible catalysis and ischemia-reperfusion injury. | ||||||
History |
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Structure visualization
Structure viewer | Molecule: ![]() ![]() |
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Downloads & links
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Download
PDBx/mmCIF format | ![]() | 2.1 MB | Display | ![]() |
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PDB format | ![]() | 1.4 MB | Display | ![]() |
PDBx/mmJSON format | ![]() | Tree view | ![]() | |
Others | ![]() |
-Validation report
Summary document | ![]() | 4.1 MB | Display | ![]() |
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Full document | ![]() | 4.3 MB | Display | |
Data in XML | ![]() | 251.1 KB | Display | |
Data in CIF | ![]() | 359.7 KB | Display | |
Arichive directory | ![]() ![]() | HTTPS FTP |
-Related structure data
Related structure data | ![]() 52878MC ![]() 9ihoC ![]() 9ihpC ![]() 9ihqC C: citing same article ( M: map data used to model this data |
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Similar structure data | Similarity search - Function & homology ![]() |
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Links
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Assembly
Deposited unit | ![]()
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Components
+Protein , 29 types, 29 molecules 1BCDEGIKOPRSVWXYbcdefghijklmp
-NADH-ubiquinone oxidoreductase chain ... , 6 types, 6 molecules AHJLMN
#2: Protein | Mass: 16019.821 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) ![]() References: UniProt: E1UWC4, NADH:ubiquinone reductase (H+-translocating) |
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#9: Protein | Mass: 39698.844 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) ![]() References: UniProt: E1UWC2, NADH:ubiquinone reductase (H+-translocating) |
#11: Protein | Mass: 18564.352 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) ![]() References: UniProt: E1UWC7, NADH:ubiquinone reductase (H+-translocating) |
#13: Protein | Mass: 72436.070 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) ![]() References: UniProt: E1UWC6, NADH:ubiquinone reductase (H+-translocating) |
#14: Protein | Mass: 54784.211 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) ![]() References: UniProt: E1UWC5, NADH:ubiquinone reductase (H+-translocating) |
#15: Protein | Mass: 59097.926 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) ![]() References: UniProt: E1UWC3, NADH:ubiquinone reductase (H+-translocating) |
-NADH dehydrogenase [ubiquinone] ... , 7 types, 7 molecules FQZanoq
#7: Protein | Mass: 52234.660 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) ![]() References: UniProt: E1UWA7, NADH:ubiquinone reductase (H+-translocating) |
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#18: Protein | Mass: 18241.705 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) ![]() |
#27: Protein | Mass: 17243.695 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) ![]() |
#28: Protein | Mass: 17446.691 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) ![]() |
#41: Protein | Mass: 12845.718 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) ![]() |
#42: Protein | Mass: 10473.039 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) ![]() |
#44: Protein | Mass: 16331.366 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) ![]() |
-Acyl carrier ... , 2 types, 2 molecules TU
#21: Protein | Mass: 15507.594 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) ![]() |
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#22: Protein | Mass: 14780.592 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) ![]() |
-Non-polymers , 10 types, 61 molecules 


















#45: Chemical | ChemComp-PLC / #46: Chemical | ChemComp-SF4 / #47: Chemical | #48: Chemical | ChemComp-FMN / | #49: Chemical | ChemComp-K / | #50: Chemical | ChemComp-3PE / #51: Chemical | ChemComp-CDL / #52: Chemical | ChemComp-NDP / | #53: Chemical | ChemComp-ZN / | #54: Chemical | |
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-Details
Has ligand of interest | N |
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Has protein modification | Y |
-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: Pichia Pastoris (Komagataella phaffi) mitochondrial complex I (NADH:ubiquinone oxidoreductase) Type: COMPLEX / Entity ID: #1-#44 / Source: NATURAL | |||||||||||||||
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Molecular weight | Value: 0.926 MDa / Experimental value: YES | |||||||||||||||
Source (natural) | Organism: ![]() | |||||||||||||||
Buffer solution | pH: 7.5 Details: 10 mM MOPS, 50 mM KCl, pH 7.5 at 4 degrees celsius. | |||||||||||||||
Buffer component |
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Specimen | Conc.: 0.7 mg/ml / Embedding applied: NO / Shadowing applied: NO / Staining applied: NO / Vitrification applied: YES Details: LMNG-solubilised Pichia Pastoris CI reconstituted in to nanodiscs (8:1:1 ratio of DOPC:DOPE:CDL [0.5 mg total] and 20 nM final Q10) | |||||||||||||||
Specimen support | Details: glow discharge 90s 20mA + chloroform and ethanol clean. Glow discharge 2X 90s 20mA, flipping in between. Grid material: COPPER / Grid mesh size: 300 divisions/in. / Grid type: Quantifoil R0.6/1 | |||||||||||||||
Vitrification | Instrument: FEI VITROBOT MARK III / Cryogen name: ETHANE / Humidity: 100 % / Chamber temperature: 277.15 K / Details: 2s blot time and -10 blot force |
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Electron microscopy imaging
Experimental equipment | ![]() Model: Titan Krios / Image courtesy: FEI Company |
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Microscopy | Model: TFS KRIOS |
Electron gun | Electron source: ![]() |
Electron lens | Mode: BRIGHT FIELD / Nominal magnification: 130000 X / Nominal defocus max: 1800 nm / Nominal defocus min: 600 nm / Cs: 2.7 mm / C2 aperture diameter: 50 µm |
Specimen holder | Cryogen: NITROGEN / Specimen holder model: FEI TITAN KRIOS AUTOGRID HOLDER |
Image recording | Average exposure time: 5.85 sec. / Electron dose: 47.79 e/Å2 / Film or detector model: TFS FALCON 4i (4k x 4k) / Num. of grids imaged: 1 / Num. of real images: 7574 |
EM imaging optics | Energyfilter name: TFS Selectris X / Energyfilter slit width: 10 eV |
Image scans | Width: 4096 / Height: 4096 |
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Processing
EM software | Name: PHENIX / Version: 1.21_5207 / Category: model refinement | ||||||||||||||||||||||||
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CTF correction | Type: PHASE FLIPPING ONLY | ||||||||||||||||||||||||
Particle selection | Num. of particles selected: 1350752 | ||||||||||||||||||||||||
Symmetry | Point symmetry: C1 (asymmetric) | ||||||||||||||||||||||||
3D reconstruction | Resolution: 2.8 Å / Resolution method: FSC 0.143 CUT-OFF / Num. of particles: 53716 / Num. of class averages: 1 / Symmetry type: POINT | ||||||||||||||||||||||||
Atomic model building | Space: REAL Details: The initial model was built into a map using ModelAngelo in RELION v5.0. This was manually inspected in Coot and final refinements with ligand restraints imposed performed in PHENIX v.1.21_5207 | ||||||||||||||||||||||||
Atomic model building | Details: Using ModelAngelo in RELION v5.0 / Source name: Other / Type: in silico model | ||||||||||||||||||||||||
Refinement | Cross valid method: NONE Stereochemistry target values: GeoStd + Monomer Library + CDL v1.2 | ||||||||||||||||||||||||
Displacement parameters | Biso mean: 40.87 Å2 | ||||||||||||||||||||||||
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