Cryo-EM reveals distinct conformations of E. coli ATP synthase on exposure to ATP

Abstract
Data availability
Article and author information
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Abstract

ATP synthase produces the majority of cellular energy in most cells. We have previously reported cryo-EM maps of autoinhibited E. coli ATP synthase imaged without addition of nucleotide (Sobti et al. 2016), indicating that the subunit ε engages the α, β and γ subunits to lock the enzyme and prevent functional rotation. Here we present multiple cryo-EM reconstructions of the enzyme frozen after the addition of MgATP to identify the changes that occur when this ε inhibition is removed. The maps generated show that, after exposure to MgATP, E. coli ATP synthase adopts a different conformation with a catalytic subunit changing conformation substantially and the ε C-terminal domain transitioning via an intermediate 'half-up' state to a condensed 'down' state. This work provides direct evidence for unique conformational states that occur in E. coli ATP synthase when ATP binding prevents the ε C-terminal domain from entering the inhibitory 'up' state.

Data availability

Cryo-EM maps have been deposited to the EMDB. Codes: EMD-9345, EMD-9346 and EMD-9348.

The following data sets were generated

1. Sobti M
2. Stewart AG
(2019) E. coli ATP synthase after incubation with ATP - State A
Electron Microscopy Data Bank, EMD-9345.

http://www.ebi.ac.uk/pdbe/entry/emdb/EMD-9345
1. Sobti M
2. Stewart AG
(2019) E. coli ATP synthase after incubation with ATP - State B
Electron Microscopy Data Bank, EMD-9346.

http://www.ebi.ac.uk/pdbe/entry/emdb/EMD-9346
1. Sobti M
2. Stewart AG
(2019) E. coli ATP synthase after incubation with ATP - State C
Electron Microscopy Data Bank, EMD-9348.

http://www.ebi.ac.uk/pdbe/entry/emdb/EMD-9348

Article and author information

Author details

Meghna Sobti

Molecular, Structural and Computational Biology Division, The Victor Chang Cardiac Research Institute, Darlinghurst, Australia

Competing interests
The authors declare that no competing interests exist.
Robert Ishmukhametov

Department of Physics, Clarendon Laboratory, University of Oxford, Oxford, United Kingdom

Competing interests
The authors declare that no competing interests exist.
James C Bouwer

Molecular Horizons, The University of Wollongong, Wollongong, Australia

Competing interests
The authors declare that no competing interests exist.
Anita Ayer

Vascular Biology Division, The Victor Chang Cardiac Research Institute, Darlinghurst, Australia

Competing interests
The authors declare that no competing interests exist.
Cacang Suarna

Vascular Biology Division, The Victor Chang Cardiac Research Institute, Darlinghurst, Australia

Competing interests
The authors declare that no competing interests exist.
Nicola J Smith

Molecular Cardiology and Biophysics Division, The Victor Chang Cardiac Research Institute, Darlinghurst, Australia

Competing interests
The authors declare that no competing interests exist.
Mary Christie

Molecular, Structural and Computational Biology Division, The Victor Chang Cardiac Research Institute, Darlinghurst, Australia

Competing interests
The authors declare that no competing interests exist.
Roland Stocker

Vascular Biology Division, The Victor Chang Cardiac Research Institute, Darlinghurst, Australia

Competing interests
The authors declare that no competing interests exist.
Thomas M Duncan

Department of Biochemistry and Molecular Biology, SUNY Upstate Medical University, Syracuse, United States

Competing interests
The authors declare that no competing interests exist.
Alastair G Stewart

Molecular, Structural and Computational Biology Division, The Victor Chang Cardiac Research Institute, Darlinghurst, Australia

For correspondence
a.stewart@victorchang.edu.au

Competing interests
The authors declare that no competing interests exist.

"This ORCID iD identifies the author of this article:" 0000-0002-2070-6030

Funding

National Health and Medical Research Council (APP1159347)

Alastair G Stewart

National Health and Medical Research Council (APP1146403)

Meghna Sobti
Alastair G Stewart

Australian Research Council (DE160100608)

Mary Christie

The funders had no role in study design, data collection and interpretation, or the decision to submit the work for publication.

Copyright

This article is distributed under the terms of the Creative Commons Attribution License permitting unrestricted use and redistribution provided that the original author and source are credited.