English
 
Help Privacy Policy Disclaimer
  Advanced SearchBrowse

Item

ITEM ACTIONSEXPORT
  Visualization of ATP Synthase Dimers in Mitochondria by Electron Cryo-tomography

Davies, K. M., Daum, B., Gold, V. A. M., Mühleip, A. W., Brandt, T., Blum, T., et al. (2014). Visualization of ATP Synthase Dimers in Mitochondria by Electron Cryo-tomography. Journal of Visualized Experiments, (91 doi: 10.3791/51228): e51228. doi:10.3791/51228.

Item is

Files

show Files

Locators

show

Creators

show
hide
 Creators:
Davies, Karen M.1, Author           
Daum, Bertram1, Author           
Gold, Vicky A. M.1, Author           
Mühleip, Alexander W.2, Author
Brandt, Tobias1, Author           
Blum, Thorsten1, Author           
Mills, Deryck J.1, Author           
Kühlbrandt, Werner1, Author           
Affiliations:
1Department of Structural Biology, Max Planck Institute of Biophysics, Max Planck Society, ou_2068291              
2Max Planck Society, ou_persistent13              

Content

show
hide
Free keywords: Structural Biology; electron microscopy; electron cryo-tomography; mitochondria; ultrastructure; membrane structure; membrane protein complexes; ATP synthase; energy conversion; bioenergetics
 Abstract: Electron cryo-tomography is a powerful tool in structural biology, capable of visualizing the three-dimensional structure of biological samples, such as cells, organelles, membrane vesicles, or viruses at molecular detail. To achieve this, the aqueous sample is rapidly vitrified in liquid ethane, which preserves it in a close-to-native, frozen-hydrated state. In the electron microscope, tilt series are recorded at liquid nitrogen temperature, from which 3D tomograms are reconstructed. The signal-to-noise ratio of the tomographic volume is inherently low. Recognizable, recurring features are enhanced by subtomogram averaging, by which individual subvolumes are cut out, aligned and averaged to reduce noise. In this way, 3D maps with a resolution of 2 nm or better can be obtained. A fit of available high-resolution structures to the 3D volume then produces atomic models of protein complexes in their native environment. Here we show how we use electron cryo-tomography to study the in situ organization of large membrane protein complexes in mitochondria. We find that ATP synthases are organized in rows of dimers along highly curved apices of the inner membrane cristae, whereas complex I is randomly distributed in the membrane regions on either side of the rows. By subtomogram averaging we obtained a structure of the mitochondrial ATP synthase dimer within the cristae membrane.

Details

show
hide
Language(s): eng - English
 Dates: 2014
 Publication Status: Published in print
 Pages: -
 Publishing info: -
 Table of Contents: -
 Rev. Type: Peer
 Identifiers: eDoc: 695911
DOI: 10.3791/51228
 Degree: -

Event

show

Legal Case

show

Project information

show

Source 1

show
hide
Title: Journal of Visualized Experiments
  Other : Journal of Visualized Experiments: JoVE
  Abbreviation : J. Vis. Exp.
Source Genre: Journal
 Creator(s):
Affiliations:
Publ. Info: Rockville Pike, Bethesda MD : JoVE
Pages: - Volume / Issue: (91 doi: 10.3791/51228) Sequence Number: e51228 Start / End Page: - Identifier: ISSN: 1940-087X
CoNE: https://pure.mpg.de/cone/journals/resource/1940087X