|Entry||Database: EMDB / ID: EMD-0151|
|Title||Rhesus macaque mitochondrial complex I|
|Sample||Rhesus macaque mitochondrial complex I|
|Biological species||Macaca mulatta (Rhesus monkey)|
|Method||single particle reconstruction / cryo EM / Resolution: 4.1 Å|
|Authors||Agip ANA / Blaza JN / Fedor JG / Hirst J|
|Funding support|| United Kingdom, 1 items |
|Citation||Journal: Annu Rev Biophys / Year: 2019|
Title: Mammalian Respiratory Complex I Through the Lens of Cryo-EM.
Authors: Ahmed-Noor A Agip / James N Blaza / Justin G Fedor / Judy Hirst /
Abstract: Single-particle electron cryomicroscopy (cryo-EM) has led to a revolution in structural work on mammalian respiratory complex I. Complex I (mitochondrial NADH:ubiquinone oxidoreductase), a membrane- ...Single-particle electron cryomicroscopy (cryo-EM) has led to a revolution in structural work on mammalian respiratory complex I. Complex I (mitochondrial NADH:ubiquinone oxidoreductase), a membrane-bound redox-driven proton pump, is one of the largest and most complicated enzymes in the mammalian cell. Rapid progress, following the first 5-Å resolution data on bovine complex I in 2014, has led to a model for mouse complex I at 3.3-Å resolution that contains 96% of the 8,518 residues and to the identification of different particle classes, some of which are assigned to biochemically defined states. Factors that helped improve resolution, including improvements to biochemistry, cryo-EM grid preparation, data collection strategy, and image processing, are discussed. Together with recent structural data from an ancient relative, membrane-bound hydrogenase, cryo-EM on mammalian complex I has provided new insights into the proton-pumping machinery and a foundation for understanding the enzyme's catalytic mechanism.
|Structure viewer||EM map: |
Downloads & links
|File||Download / File: emd_0151.map.gz / Format: CCP4 / Size: 178 MB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES)|
|Projections & slices|
Images are generated by Spider.
|Voxel size||X=Y=Z: 1.39 Å|
|Symmetry||Space group: 1|
CCP4 map header:
|Projections & Slices|
-Half map: #1
|Projections & Slices|
-Half map: #2
-Entire Rhesus macaque mitochondrial complex I
|Entire||Name: Rhesus macaque mitochondrial complex I / Number of Components: 1|
-Component #1: protein, Rhesus macaque mitochondrial complex I
|Protein||Name: Rhesus macaque mitochondrial complex I / Recombinant expression: No|
|Mass||Theoretical: 1000 kDa|
|Source||Species: Macaca mulatta (Rhesus monkey)|
|Source (natural)||Organelle: Mitochondria / Organ Or Tissue: Heart|
|Specimen||Specimen State: Particle / Method: cryo EM|
|Sample solution||Specimen conc.: 3 mg/mL / pH: 7.14|
|Support film||UltrAuFoil gold grids (0.6/1) was used for the preparation of macaque complex I grids. In brief, UltrAuFoil gold grids (0.6/1) were glow discharged at 20 mA for 60 sec using a EMITECH glow-discharger unit. Then, the grids were incubated in 5 mM 11-mercaptoundecyl hexaethyleneglycol (SPT-0011P6, SensoPath Technologies) in ethanol for two days under anaerobic conditions.|
|Vitrification||Instrument: FEI VITROBOT MARK IV / Cryogen Name: ETHANE / Temperature: 277 K / Humidity: 100 % / Details: 8-16s blotting..|
-Electron microscopy imaging
Model: Titan Krios / Image courtesy: FEI Company
|Imaging||Microscope: FEI TITAN KRIOS|
|Electron gun||Electron Source: FIELD EMISSION GUN / Accelerating Voltage: 300 kV / Electron Dose: 50 e/Å2 / Illumination Mode: FLOOD BEAM|
|Lens||Magnification: 59000 X (nominal) / Cs: 2.7 mm / Imaging Mode: BRIGHT FIELD|
|Specimen Holder||Model: FEI TITAN KRIOS AUTOGRID HOLDER|
|Camera||Detector: FEI FALCON II (4k x 4k)|
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