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- EMDB-52877: Closed state with NUQM and without flavoprotein (classification s... -
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Open data
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Basic information
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Title | Closed state with NUQM and without flavoprotein (classification state 2) of Pichia pastoris mitochondrial complex I in cMSP26 nanodiscs | |||||||||
![]() | Consensus refinement map postprocessed with RELION and used for model building and model refinement | |||||||||
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![]() | NADH:ubiquinone oxidoreductase nanodisc membrane protein complex / MEMBRANE PROTEIN | |||||||||
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 / NAD 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 | single particle reconstruction / cryo EM / Resolution: 2.9 Å | |||||||||
![]() | Grba DN / 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
Supplemental images |
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Downloads & links
-EMDB archive
Map data | ![]() | 556 MB | ![]() | |
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Header (meta data) | ![]() ![]() | 68 KB 68 KB | Display Display | ![]() |
FSC (resolution estimation) | ![]() | 19.1 KB | Display | ![]() |
Images | ![]() | 153.7 KB | ||
Masks | ![]() | 600.7 MB | ![]() | |
Filedesc metadata | ![]() | 15 KB | ||
Others | ![]() ![]() | 487 MB 487.2 MB | ||
Archive directory | ![]() ![]() | HTTPS FTP |
-Validation report
Summary document | ![]() | 1.3 MB | Display | ![]() |
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Full document | ![]() | 1.3 MB | Display | |
Data in XML | ![]() | 25.5 KB | Display | |
Data in CIF | ![]() | 33.8 KB | Display | |
Arichive directory | ![]() ![]() | HTTPS FTP |
-Related structure data
Related structure data | ![]() 9ihqMC ![]() 9ihoC ![]() 9ihpC ![]() 9ihrC M: atomic model generated by this map C: citing same article ( |
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Similar structure data | Similarity search - Function & homology ![]() |
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Links
EMDB pages | ![]() ![]() |
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Related items in Molecule of the Month |
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Map
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Annotation | Consensus refinement map postprocessed with RELION and used for model building and model refinement | ||||||||||||||||||||||||||||||||||||
Projections & slices | Image control
Images are generated by Spider. | ||||||||||||||||||||||||||||||||||||
Voxel size | X=Y=Z: 0.929 Å | ||||||||||||||||||||||||||||||||||||
Density |
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Symmetry | Space group: 1 | ||||||||||||||||||||||||||||||||||||
Details | EMDB XML:
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-Supplemental data
-Mask #1
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Density Histograms |
-Half map: Half map 2
File | emd_52877_half_map_1.map | ||||||||||||
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Annotation | Half map 2 | ||||||||||||
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Density Histograms |
-Half map: Half map 1
File | emd_52877_half_map_2.map | ||||||||||||
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Annotation | Half map 1 | ||||||||||||
Projections & Slices |
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Density Histograms |
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Sample components
+Entire : Pichia Pastoris (Komagataella phaffi) mitochondrial complex I (NA...
+Supramolecule #1: Pichia Pastoris (Komagataella phaffi) mitochondrial complex I (NA...
+Macromolecule #1: Altered inheritance of mitochondria protein 41, mitochondrial
+Macromolecule #2: NADH-ubiquinone oxidoreductase chain 3
+Macromolecule #3: BA75_00622T0
+Macromolecule #4: NUGM (30 kDa) subunit of mitochondrial NADH:ubiquinone oxidoreduc...
+Macromolecule #5: NUCM (49 kDa) subunit of mitochondrial NADH:ubiquinone oxidoreduc...
+Macromolecule #6: NUAM (75 kDa) subunit of mitochondrial NADH:ubiquinone oxidoreduc...
+Macromolecule #7: NADH-ubiquinone oxidoreductase chain 1
+Macromolecule #8: NUIM (TYKY) subunit of mitochondrial NADH:ubiquinone oxidoreducta...
+Macromolecule #9: NADH-ubiquinone oxidoreductase chain 6
+Macromolecule #10: NULM (ND4L) subunit of mitochondrial NADH:ubiquinone oxidoreducta...
+Macromolecule #11: NADH-ubiquinone oxidoreductase chain 5
+Macromolecule #12: NADH-ubiquinone oxidoreductase chain 4
+Macromolecule #13: NADH-ubiquinone oxidoreductase chain 2
+Macromolecule #14: NUXM subunit of mitochondrial NADH:ubiquinone oxidoreductase (Com...
+Macromolecule #15: NUEM (39 kDa) subunit of mitochondrial NADH:ubiquinone oxidoreduc...
+Macromolecule #16: NADH dehydrogenase [ubiquinone] iron-sulfur protein 4, mitochondrial
+Macromolecule #17: NUMM (13 kDa) subunit of mitochondrial NADH:ubiquinone oxidoreduc...
+Macromolecule #18: NI8M (B8) subunit of mitochondrial NADH:ubiquinone oxidoreductase...
+Macromolecule #19: Acyl carrier protein
+Macromolecule #20: Acyl carrier protein
+Macromolecule #21: NUFM (B13) subunit of mitochondrial NADH:ubiquinone oxidoreductas...
+Macromolecule #22: BA75_04796T0
+Macromolecule #23: NADH-ubiquinone oxidoreductase
+Macromolecule #24: NUJM (B14.7) subunit of mitochondrial NADH:ubiquinone oxidoreduct...
+Macromolecule #25: NADH dehydrogenase [ubiquinone] 1 alpha subcomplex subunit 13
+Macromolecule #26: NADH dehydrogenase [ubiquinone] 1 alpha subcomplex subunit 1
+Macromolecule #27: NI9M (B9) subunit of mitochondrial NADH:ubiquinone oxidoreductase...
+Macromolecule #28: BA75_00589T0
+Macromolecule #29: Pichia pastoris NADH-ubiquinone oxidoreductase subunit NEBM
+Macromolecule #30: BA75_05084T0
+Macromolecule #31: NUTM subunit of mitochondrial NADH:ubiquinone oxidoreductase (Com...
+Macromolecule #32: NESM (ESSS) subunit of mitochondrial NADH:ubiquinone oxidoreducta...
+Macromolecule #33: NUSM subunit of mitochondrial NADH:ubiquinone oxidoreductase (Com...
+Macromolecule #34: NUUM subunit of mitochondrial NADH:ubiquinone oxidoreductase (Com...
+Macromolecule #35: Subunit of mitochondrial NADH:ubiquinone oxidoreductase (Complex I)
+Macromolecule #36: NB2M (B12) subunit of mitochondrial NADH:ubiquinone oxidoreductas...
+Macromolecule #37: NIAM (ASHI) subunit of mitochondrial NADH:ubiquinone oxidoreducta...
+Macromolecule #38: NB5M (B15) subunit of mitochondrial NADH:ubiquinone oxidoreductas...
+Macromolecule #39: NADH dehydrogenase [ubiquinone] 1 beta subcomplex subunit 9
+Macromolecule #40: NADH dehydrogenase [ubiquinone] 1 beta subcomplex subunit 7
+Macromolecule #41: NIDM (PDSW) subunit of mitochondrial NADH:ubiquinone oxidoreducta...
+Macromolecule #42: NADH dehydrogenase [ubiquinone] 1 alpha subcomplex subunit
+Macromolecule #43: DIUNDECYL PHOSPHATIDYL CHOLINE
+Macromolecule #44: IRON/SULFUR CLUSTER
+Macromolecule #45: FE2/S2 (INORGANIC) CLUSTER
+Macromolecule #46: POTASSIUM ION
+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...
-Experimental details
-Structure determination
Method | cryo EM |
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![]() | single particle reconstruction |
Aggregation state | particle |
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Sample preparation
Concentration | 0.7 mg/mL | |||||||||
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Buffer | pH: 7.5 Component:
Details: 10 mM MOPS, 50 mM KCl, pH 7.5 at 4 degrees celsius. | |||||||||
Grid | Model: Quantifoil R1.2/1.3 / Material: COPPER / Support film - Material: CARBON / Support film - topology: HOLEY / Pretreatment - Type: GLOW DISCHARGE / Pretreatment - Time: 180 sec. / Pretreatment - Atmosphere: AIR / Pretreatment - Pressure: 0.039 kPa Details: glow discharged for 2X 90s, flipping the grid in between | |||||||||
Vitrification | Cryogen name: ETHANE / Chamber humidity: 100 % / Chamber temperature: 277.15 K / Instrument: FEI VITROBOT MARK III / Details: 2s blot time and -10 blot force. | |||||||||
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) |
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Electron microscopy
Microscope | TFS KRIOS |
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Specialist optics | Energy filter - Name: TFS Selectris X / Energy filter - Slit width: 10 eV |
Image recording | Film or detector model: TFS FALCON 4i (4k x 4k) / Digitization - Dimensions - Width: 4096 pixel / Digitization - Dimensions - Height: 4096 pixel / Number grids imaged: 1 / Number real images: 7574 / Average exposure time: 5.85 sec. / Average electron dose: 47.79 e/Å2 |
Electron beam | Acceleration voltage: 300 kV / Electron source: ![]() |
Electron optics | C2 aperture diameter: 50.0 µm / Illumination mode: FLOOD BEAM / Imaging mode: BRIGHT FIELD / Cs: 2.7 mm / Nominal defocus max: 1.8 µm / Nominal defocus min: 0.6 µm / Nominal magnification: 130000 |
Sample stage | Specimen holder model: FEI TITAN KRIOS AUTOGRID HOLDER / Cooling holder cryogen: NITROGEN |
Experimental equipment | ![]() Model: Titan Krios / Image courtesy: FEI Company |
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Image processing
-Atomic model buiding 1
Initial model | PDB ID: Chain - Source name: PDB / Chain - Initial model type: experimental model / Details: State 1 from same data |
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Details | The initial model was made for state 1 and then subunits removed to match this state. This was manually inspected in Coot and final refinements with ligand restraints imposed performed in PHENIX v.1.21_5207 |
Refinement | Space: REAL |
Output model | ![]() PDB-9ihq: |