Arechaga, Ignacio; Fotiadis, Dimitrios (2007). Reconstitution of mitochondrial ATP synthase into lipid bilayers for structural analysis. Journal of structural biology, 160(3), pp. 287-94. San Diego, Calif.: Elsevier 10.1016/j.jsb.2007.09.007
Full text not available from this repository.Mitochondrial F(1)F(o)-ATP synthase is a molecular motor that couples the energy generated by oxidative metabolism to the synthesis of ATP. Direct visualization of the rotary action of the bacterial ATP synthase has been well characterized. However, direct observation of rotation of the mitochondrial enzyme has not been reported yet. Here, we describe two methods to reconstitute mitochondrial F(1)F(o)-ATP synthase into lipid bilayers suitable for structure analysis by electron and atomic force microscopy (AFM). Proteoliposomes densely packed with bovine heart mitochondria F(1)F(o)-ATP synthase were obtained upon detergent removal from ternary mixtures (lipid, detergent and protein). Two-dimensional crystals of recombinant hexahistidine-tagged yeast F(1)F(o)-ATP synthase were grown using the supported monolayer technique. Because the hexahistidine-tag is located at the F(1) catalytic subcomplex, ATP synthases were oriented unidirectionally in such two-dimensional crystals, exposing F(1) to the lipid monolayer and the F(o) membrane region to the bulk solution. This configuration opens a new avenue for the determination of the c-ring stoichiometry of unknown hexahistidine-tagged ATP synthases and the organization of the membrane intrinsic subunits within F(o) by electron microscopy and AFM.
Item Type: |
Journal Article (Original Article) |
|---|---|
Division/Institute: |
04 Faculty of Medicine > Pre-clinic Human Medicine > Institute of Biochemistry and Molecular Medicine |
UniBE Contributor: |
Fotiadis, Dimitrios José |
ISSN: |
1047-8477 |
ISBN: |
17959389 |
Publisher: |
Elsevier |
Language: |
English |
Submitter: |
Factscience Import |
Date Deposited: |
04 Oct 2013 14:53 |
Last Modified: |
05 Dec 2022 14:16 |
Publisher DOI: |
10.1016/j.jsb.2007.09.007 |
PubMed ID: |
17959389 |
Web of Science ID: |
000251473500004 |
URI: |
https://boris.unibe.ch/id/eprint/22700 (FactScience: 36075) |
Actions (login required)
 |
Edit item |