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          Institute: MPI für Biophysik     Collection: Abt. Strukturbiologie     Display Documents

ID: 695911.0, MPI für Biophysik / Abt. Strukturbiologie
Visualization of ATP Synthase Dimers in Mitochondria by Electron Cryo-tomography
Authors:Davies, Karen M.; Daum, Bertram; Gold, Vicky A. M.; Mühleip, Alexander W.; Brandt, Tobias; Blum, Thorsten; Mills, Deryck J.; Kühlbrandt, Werner
Date of Publication (YYYY-MM-DD):2014
Title of Journal:The Journal of Visualized Experiments
Journal Abbrev.:JoVE
Issue / Number:91 doi: 10.3791/51228
Sequence Number of Article:e51228
Review Status:Peer-review
Audience:Experts Only
Abstract / Description: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.
Free Keywords:Structural Biology; electron microscopy; electron cryo-tomography; mitochondria; ultrastructure; membrane structure; membrane protein complexes; ATP synthase; energy conversion; bioenergetics
External Publication Status:published
Document Type:Article
Communicated by:N. N.
Affiliations:MPI für Biophysik/Abteilung Strukturbiologie
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