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Yorodumi- PDB-7ak5: Cryo-EM structure of respiratory complex I in the deactive state ... -
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| Entry | Database: PDB / ID: 7ak5 | |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
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| Title | Cryo-EM structure of respiratory complex I in the deactive state from Mus musculus at 3.2 A | |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
|  Components | 
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|  Keywords | OXIDOREDUCTASE / deactive complex I | |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
| Function / homology |  Function and homology information Mitochondrial protein import / response to injury involved in regulation of muscle adaptation / Protein lipoylation / Mitochondrial Fatty Acid Beta-Oxidation / Complex I biogenesis / RHOG GTPase cycle / Respiratory electron transport / protein insertion into mitochondrial inner membrane / blastocyst hatching / respiratory system process ...Mitochondrial protein import / response to injury involved in regulation of muscle adaptation / Protein lipoylation / Mitochondrial Fatty Acid Beta-Oxidation / Complex I biogenesis / RHOG GTPase cycle / Respiratory electron transport / protein insertion into mitochondrial inner membrane / blastocyst hatching / respiratory system process / psychomotor behavior / Mitochondrial protein degradation / cellular response to oxygen levels / response to light intensity / mesenchymal stem cell proliferation / respiratory chain complex / reproductive system development / mitochondrial [2Fe-2S] assembly complex / iron-sulfur cluster assembly complex / mitochondrial large ribosomal subunit binding / gliogenesis / mesenchymal stem cell differentiation / circulatory system development / negative regulation of non-canonical NF-kappaB signal transduction / cardiac muscle tissue development / neural precursor cell proliferation / [2Fe-2S] cluster assembly / adult walking behavior / oxygen sensor activity / positive regulation of mitochondrial membrane potential / response to hydroperoxide / cellular response to glucocorticoid stimulus / stem cell division / NADH dehydrogenase activity / iron-sulfur cluster assembly / mitochondrial ATP synthesis coupled electron transport / dopamine metabolic process / NADH:ubiquinone reductase (H+-translocating) / adult behavior / positive regulation of ATP biosynthetic process / mitochondrial respiratory chain complex I assembly / mitochondrial electron transport, NADH to ubiquinone / proton motive force-driven mitochondrial ATP synthesis / respiratory chain complex I / positive regulation of execution phase of apoptosis / NADH dehydrogenase (ubiquinone) activity / neuron development / quinone binding / cellular response to interferon-beta / ATP synthesis coupled electron transport / negative regulation of reactive oxygen species biosynthetic process / extrinsic apoptotic signaling pathway / cellular response to retinoic acid / tricarboxylic acid cycle / neurogenesis / Neutrophil degranulation / visual perception / muscle contraction / reactive oxygen species metabolic process / aerobic respiration / cerebellum development / regulation of mitochondrial membrane potential / respiratory electron transport chain / response to nicotine / response to cocaine / mitochondrion organization / DNA damage response, signal transduction by p53 class mediator / kidney development / response to hydrogen peroxide / sensory perception of sound / monooxygenase activity / fatty acid metabolic process / electron transport chain / circadian rhythm / brain development / mitochondrial membrane / mitochondrial intermembrane space / 2 iron, 2 sulfur cluster binding / cognition / multicellular organism growth / NAD binding / positive regulation of protein catabolic process / fatty acid biosynthetic process / cellular senescence / FMN binding / nervous system development / myelin sheath / 4 iron, 4 sulfur cluster binding / response to oxidative stress / neuron apoptotic process / response to ethanol / gene expression / in utero embryonic development / response to hypoxia / electron transfer activity / mitochondrial inner membrane / nuclear speck / nuclear body / mitochondrial matrix / response to xenobiotic stimulus Similarity search - Function | |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
| Biological species |   Mus musculus (house mouse) | |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
| Method | ELECTRON MICROSCOPY / single particle reconstruction / cryo EM / Resolution: 3.17 Å | |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
|  Authors | Yin, Z. / Bridges, H.R. / Grba, D. / Hirst, J. | |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
| Funding support |  United Kingdom, 2items 
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|  Citation |  Journal: Nat Commun / Year: 2021 Title: Structural basis for a complex I mutation that blocks pathological ROS production. Authors: Zhan Yin / Nils Burger / Duvaraka Kula-Alwar / Dunja Aksentijević / Hannah R Bridges / Hiran A Prag / Daniel N Grba / Carlo Viscomi / Andrew M James / Amin Mottahedin / Thomas Krieg / ...Authors: Zhan Yin / Nils Burger / Duvaraka Kula-Alwar / Dunja Aksentijević / Hannah R Bridges / Hiran A Prag / Daniel N Grba / Carlo Viscomi / Andrew M James / Amin Mottahedin / Thomas Krieg / Michael P Murphy / Judy Hirst /      Abstract: Mitochondrial complex I is central to the pathological reactive oxygen species (ROS) production that underlies cardiac ischemia-reperfusion (IR) injury. ND6-P25L mice are homoplasmic for a disease- ...Mitochondrial complex I is central to the pathological reactive oxygen species (ROS) production that underlies cardiac ischemia-reperfusion (IR) injury. ND6-P25L mice are homoplasmic for a disease-causing mtDNA point mutation encoding the P25L substitution in the ND6 subunit of complex I. The cryo-EM structure of ND6-P25L complex I revealed subtle structural changes that facilitate rapid conversion to the "deactive" state, usually formed only after prolonged inactivity. Despite its tendency to adopt the "deactive" state, the mutant complex is fully active for NADH oxidation, but cannot generate ROS by reverse electron transfer (RET). ND6-P25L mitochondria function normally, except for their lack of RET ROS production, and ND6-P25L mice are protected against cardiac IR injury in vivo. Thus, this single point mutation in complex I, which does not affect oxidative phosphorylation but renders the complex unable to catalyse RET, demonstrates the pathological role of ROS production by RET during IR injury. | |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
| History | 
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- Structure visualization
Structure visualization
| Movie | 
 
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| Structure viewer | Molecule:  Molmil  Jmol/JSmol | 
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Download
| PDBx/mmCIF format |  7ak5.cif.gz | 1.5 MB | Display |  PDBx/mmCIF format | 
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| PDB format |  pdb7ak5.ent.gz | 1.2 MB | Display |  PDB format | 
| PDBx/mmJSON format |  7ak5.json.gz | Tree view |  PDBx/mmJSON format | |
| Others |  Other downloads | 
-Validation report
| Summary document |  7ak5_validation.pdf.gz | 2.7 MB | Display |  wwPDB validaton report | 
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| Full document |  7ak5_full_validation.pdf.gz | 2.7 MB | Display | |
| Data in XML |  7ak5_validation.xml.gz | 208.6 KB | Display | |
| Data in CIF |  7ak5_validation.cif.gz | 315.1 KB | Display | |
| Arichive directory |  https://data.pdbj.org/pub/pdb/validation_reports/ak/7ak5  ftp://data.pdbj.org/pub/pdb/validation_reports/ak/7ak5 | HTTPS FTP | 
-Related structure data
| Related structure data |  11810MC  7ak6C M: map data used to model this data C: citing same article ( | 
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| Similar structure data | |
| EM raw data |  EMPIAR-10604 (Title: Single particle cryo-EM dataset of Mus musculus mitochondrial complex I in the deactive state Data size: 2.7 TB Data #1: Single particle cryo-EM dataset of Mus musculus mitochondrial complex I in the deactive state [micrographs - multiframe]) | 
- Links
Links
- Assembly
Assembly
| Deposited unit |  
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| 1 | 
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- Components
Components
-NADH-ubiquinone oxidoreductase chain  ... , 7 types, 7 molecules AHJKLMN      
| #1: Protein | Mass: 13251.785 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural)    Mus musculus (house mouse) / Tissue: heart References: UniProt: P03899, NADH:ubiquinone reductase (H+-translocating) | 
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| #8: Protein | Mass: 36105.027 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural)    Mus musculus (house mouse) / Tissue: heart References: UniProt: P03888, NADH:ubiquinone reductase (H+-translocating) | 
| #10: Protein | Mass: 18656.100 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural)    Mus musculus (house mouse) / Tissue: heart References: UniProt: P03925, NADH:ubiquinone reductase (H+-translocating) | 
| #11: Protein | Mass: 10637.629 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural)    Mus musculus (house mouse) / Tissue: heart References: UniProt: P03903, NADH:ubiquinone reductase (H+-translocating) | 
| #12: Protein | Mass: 68547.297 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural)    Mus musculus (house mouse) / Tissue: heart References: UniProt: P03921, NADH:ubiquinone reductase (H+-translocating) | 
| #13: Protein | Mass: 51943.547 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural)    Mus musculus (house mouse) / Tissue: heart References: UniProt: P03911, NADH:ubiquinone reductase (H+-translocating) | 
| #14: Protein | Mass: 38800.230 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural)    Mus musculus (house mouse) / Tissue: heart References: UniProt: P03893, NADH:ubiquinone reductase (H+-translocating) | 
-NADH dehydrogenase [ubiquinone] iron-sulfur protein  ... , 7 types, 7 molecules BCDIQRe      
| #2: Protein | Mass: 24715.912 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural)    Mus musculus (house mouse) / Tissue: heart References: UniProt: Q9DC70, NADH:ubiquinone reductase (H+-translocating) | 
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| #3: Protein | Mass: 30191.307 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural)    Mus musculus (house mouse) / Tissue: heart References: UniProt: Q9DCT2, NADH:ubiquinone reductase (H+-translocating) | 
| #4: Protein | Mass: 52720.602 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural)    Mus musculus (house mouse) / Tissue: heart References: UniProt: Q91WD5, NADH:ubiquinone reductase (H+-translocating) | 
| #9: Protein | Mass: 24068.355 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural)    Mus musculus (house mouse) / Tissue: heart References: UniProt: Q8K3J1, NADH:ubiquinone reductase (H+-translocating) | 
| #17: Protein | Mass: 19814.725 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural)    Mus musculus (house mouse) / Tissue: heart / References: UniProt: Q9CXZ1 | 
| #18: Protein | Mass: 12765.534 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural)    Mus musculus (house mouse) / Tissue: heart / References: UniProt: P52503 | 
| #30: Protein | Mass: 12675.772 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural)    Mus musculus (house mouse) / Tissue: heart / References: UniProt: Q99LY9 | 
-NADH dehydrogenase [ubiquinone] flavoprotein  ... , 3 types, 3 molecules EFs  
| #5: Protein | Mass: 27077.047 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural)    Mus musculus (house mouse) / Tissue: heart References: UniProt: Q9D6J6, NADH:ubiquinone reductase (H+-translocating) | 
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| #6: Protein | Mass: 50904.152 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural)    Mus musculus (house mouse) / Tissue: heart References: UniProt: Q91YT0, NADH:ubiquinone reductase (H+-translocating) | 
| #44: Protein | Mass: 11833.504 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural)    Mus musculus (house mouse) / Tissue: heart / References: UniProt: Q8BK30 | 
-Protein , 3 types, 4 molecules GTUY   
| #7: Protein | Mass: 78652.367 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural)    Mus musculus (house mouse) / Tissue: heart References: UniProt: Q91VD9, NADH:ubiquinone reductase (H+-translocating) | ||
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| #20: Protein | Mass: 17390.289 Da / Num. of mol.: 2 / Source method: isolated from a natural source / Source: (natural)    Mus musculus (house mouse) / Tissue: heart / References: UniProt: Q9CR21 #24: Protein |  | Mass: 15130.416 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural)    Mus musculus (house mouse) / Tissue: heart / References: UniProt: G5E814 | 
-NADH dehydrogenase [ubiquinone] 1 alpha subcomplex subunit  ... , 11 types, 11 molecules OPSVWXZabqr          
| #15: Protein | Mass: 40657.375 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural)    Mus musculus (house mouse) / Tissue: heart / References: UniProt: Q99LC3 | 
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| #16: Protein | Mass: 42588.129 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural)    Mus musculus (house mouse) / Tissue: heart / References: UniProt: Q9DC69 | 
| #19: Protein | Mass: 10932.675 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural)    Mus musculus (house mouse) / Tissue: heart / References: UniProt: Q9CQ75 | 
| #21: Protein | Mass: 13380.719 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural)    Mus musculus (house mouse) / Tissue: heart / References: UniProt: Q9CPP6 | 
| #22: Protein | Mass: 15311.858 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural)    Mus musculus (house mouse) / Tissue: heart / References: UniProt: Q9CQZ5 | 
| #23: Protein | Mass: 20025.127 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural)    Mus musculus (house mouse) / Tissue: heart / References: UniProt: Q9DCJ5 | 
| #25: Protein | Mass: 16881.588 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural)    Mus musculus (house mouse) / Tissue: heart / References: UniProt: Q9ERS2 | 
| #26: Protein | Mass: 7921.279 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural)    Mus musculus (house mouse) / Tissue: heart / References: UniProt: O35683 | 
| #27: Protein | Mass: 9249.709 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural)    Mus musculus (house mouse) / Tissue: heart / References: UniProt: Q9CQ91 | 
| #42: Protein | Mass: 17112.416 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural)    Mus musculus (house mouse) / Tissue: heart / References: UniProt: Q7TMF3 | 
| #43: Protein | Mass: 12506.432 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural)    Mus musculus (house mouse) / Tissue: heart / References: UniProt: Q9Z1P6 | 
-NADH dehydrogenase [ubiquinone] 1 subunit  ... , 2 types, 2 molecules cd 
| #28: Protein | Mass: 8636.023 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural)    Mus musculus (house mouse) / Tissue: heart / References: UniProt: Q9CQY9 | 
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| #29: Protein | Mass: 14185.692 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural)    Mus musculus (house mouse) / Tissue: heart / References: UniProt: Q9CQ54 | 
-NADH dehydrogenase [ubiquinone] 1 beta subcomplex subunit  ... , 11 types, 11 molecules fghijklmnop          
| #31: Protein | Mass: 6965.109 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural)    Mus musculus (house mouse) / Tissue: heart / References: UniProt: P0DN34 | 
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| #32: Protein | Mass: 17463.727 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural)    Mus musculus (house mouse) / Tissue: heart / References: UniProt: O09111 | 
| #33: Protein | Mass: 21742.197 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural)    Mus musculus (house mouse) / Tissue: heart / References: UniProt: Q9CQH3 | 
| #34: Protein | Mass: 15450.925 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural)    Mus musculus (house mouse) / Tissue: heart / References: UniProt: Q3UIU2 | 
| #35: Protein | Mass: 11982.437 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural)    Mus musculus (house mouse) / Tissue: heart / References: UniProt: Q9CPU2 | 
| #36: Protein | Mass: 11714.240 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural)    Mus musculus (house mouse) / Tissue: heart / References: UniProt: Q9CQZ6 | 
| #37: Protein | Mass: 21903.828 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural)    Mus musculus (house mouse) / Tissue: heart / References: UniProt: Q9D6J5 | 
| #38: Protein | Mass: 15105.287 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural)    Mus musculus (house mouse) / Tissue: heart / References: UniProt: Q9CQC7 | 
| #39: Protein | Mass: 22020.123 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural)    Mus musculus (house mouse) / Tissue: heart / References: UniProt: Q9CQJ8 | 
| #40: Protein | Mass: 16360.804 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural)    Mus musculus (house mouse) / Tissue: heart / References: UniProt: Q9CR61 | 
| #41: Protein | Mass: 21054.832 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural)    Mus musculus (house mouse) / Tissue: heart / References: UniProt: Q9DCS9 | 
-Non-polymers , 10 types, 28 molecules 


















| #45: Chemical | ChemComp-SF4 / #46: Chemical | #47: Chemical | #48: Chemical | ChemComp-FMN / | #49: Chemical | ChemComp-3PE / #50: Chemical | ChemComp-CDL / #51: Chemical | ChemComp-ATP / | #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 | 
- Sample preparation
Sample preparation
| Component | Name: Mouse mitochondrial complex I / Type: COMPLEX / Details: complex I in the deactive state / Entity ID: #1-#44 / Source: NATURAL | 
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| Molecular weight | Value: 1 MDa / Experimental value: NO | 
| Source (natural) | Organism:   Mus musculus (house mouse) / Strain: C57BL/6J / Organ: heart / Organelle: Mitochondrion | 
| Buffer solution | pH: 7.4 | 
| Specimen | Embedding applied: NO / Shadowing applied: NO / Staining applied: NO / Vitrification applied: YES | 
| Specimen support | Grid material: GOLD / Grid type: UltrAuFoil R0.6/1 | 
| Vitrification | Instrument: FEI VITROBOT MARK IV / Cryogen name: ETHANE / Humidity: 100 % / Chamber temperature: 277.15 K / Details: 8-16s blotting | 
- Electron microscopy imaging
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:  FIELD EMISSION GUN / Accelerating voltage: 300 kV / Illumination mode: FLOOD BEAM | 
| Electron lens | Mode: BRIGHT FIELD / Cs: 2.7 mm | 
| Image recording | Electron dose: 50 e/Å2 / Detector mode: COUNTING / Film or detector model: GATAN K2 QUANTUM (4k x 4k) | 
- Processing
Processing
| EM software | 
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| CTF correction | Type: PHASE FLIPPING AND AMPLITUDE CORRECTION | ||||||||||||||||||||||||||||
| 3D reconstruction | Resolution: 3.17 Å / Resolution method: FSC 0.143 CUT-OFF / Num. of particles: 50184 / Symmetry type: POINT | 
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