National Outstanding Young Scholar Science Foundation
China
National Natural Science Foundation of China
3162500065, 31030020 and 31170679
China
Citation
Journal: Cell / Year: 2016 Title: Structure of Mammalian Respiratory Supercomplex IIIIIV. Authors: Meng Wu / Jinke Gu / Runyu Guo / Yushen Huang / Maojun Yang / Abstract: The mammalian respiratory chain complexes assemble into supercomplexes (SCs) and reside in the inner mitochondrial membrane to transfer electrons and establish the proton gradient for complex V to ...The mammalian respiratory chain complexes assemble into supercomplexes (SCs) and reside in the inner mitochondrial membrane to transfer electrons and establish the proton gradient for complex V to synthesize ATP. The precise arrangement of SCs is largely unknown. Here, we report a 4.0-Å cryo-electron microscopy (cryo-EM) structure of the major SC in porcine heart, the 1.7-MDa SCIIIIIV. The complex III (CIII) dimer and complex IV (CIV) bind at the same side of the L-shaped complex I (CI). Several accessory or supernumerary subunits of CI, such as NDUFA11, NDUFB4, NDUFB8, and NDUFB9, directly contribute to the oligomerization of CI, CIII, and CIV. COX7C and COX7A of CIV attach CIV to the concave surface formed by CIII and the distal end of membrane arm of CI. The structure suggests a possible mechanism by which electrons are transferred from NADH to cytochrome c and provides a platform for future functional dissection of respiration.
History
Deposition
Aug 30, 2016
Deposition site: PDBJ / Processing site: PDBJ
Revision 1.0
Mar 29, 2017
Provider: repository / Type: Initial release
Revision 1.1
Nov 6, 2019
Group: Data collection / Other / Category: cell / em_image_scans / em_software / Item: _cell.Z_PDB / _em_software.name
Mass: 65.409 Da / Num. of mol.: 1 / Source method: obtained synthetically / Formula: Zn
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Details
Sequence details
THE SEQUENCE OF RESIDUE 1-47 OF CHAIN E IS MFLTAALRARATGFTAQWGRHVRNLHKTAVQNGAGGALFVHRDTPEN. THE ...THE SEQUENCE OF RESIDUE 1-47 OF CHAIN E IS MFLTAALRARATGFTAQWGRHVRNLHKTAVQNGAGGALFVHRDTPEN. THE AUTHOR COULD OBSERVE SOME CONNECTED RESIDUES AND ASSIGNED UNK TO THESE RESIDUES. THE AUTHOR IS NOT SURE WHCIH PART OF RESIDUE 1-47 CORRESPONDS WITH THIS PEPTIDE OF UNK. SO THE RESIDUE NUMBERS OF UNK IS MEANINGLESS.
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Experimental details
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Experiment
Experiment
Method: ELECTRON MICROSCOPY
EM experiment
Aggregation state: PARTICLE / 3D reconstruction method: single particle reconstruction
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Sample preparation
Component
Name: Respiratory supercomplex I1III2IV1 / Type: COMPLEX / Details: Supercomplex generated by complex I, III and IV. / Entity ID: #1-#67 / Source: NATURAL
Molecular weight
Value: 1.7 MDa / Experimental value: YES
Source (natural)
Organism: Sus scrofa (pig)
Buffer solution
pH: 7.4
Specimen
Embedding applied: NO / Shadowing applied: NO / Staining applied: NO / Vitrification applied: YES
Vitrification
Cryogen name: ETHANE / Humidity: 100 %
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Electron microscopy imaging
Experimental equipment
Model: Titan Krios / Image courtesy: FEI Company
Microscopy
Model: FEI TITAN KRIOS
Electron gun
Electron source: FIELD EMISSION GUN / Accelerating voltage: 300 kV / Illumination mode: OTHER
Electron lens
Mode: BRIGHT FIELDBright-field microscopy
Image recording
Electron dose: 1.25 e/Å2 / Film or detector model: FEI FALCON II (4k x 4k)
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Processing
EM software
ID
Name
Version
Category
1
EMAN
2.1
particleselection
2
AutoEMation
2
imageacquisition
4
CTFFIND
3
CTFcorrection
7
Coot
0.8.2
modelfitting
9
RELION
1.4
initialEulerassignment
10
RELION
1.4
finalEulerassignment
11
RELION
1.4
classification
12
RELION
1.4
3Dreconstruction
13
PHENIX
1.9
modelrefinement
CTF correction
Type: NONE
Symmetry
Point symmetry: C1 (asymmetric)
3D reconstruction
Resolution: 4 Å / Resolution method: FSC 0.143 CUT-OFF / Num. of particles: 161912 / Symmetry type: POINT
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