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Content archived on 2022-12-23

Nano-scale electromechanics of the ATP synthase and the cytochrome bc1 complex

Objective

The ATP synthase (FOF1) and the cytochrome bc1 are enzyme complexes that perform electro-mechano-chemical energy transduction. A peculiarity of both these nano-machines is intermediate mechanical reaction steps by large-scale rotation of subunits against each other.

It is planned to combine
(i) detection in real-time of the single enzyme rotary motion by microvideographic and spectroscopic assays;
(ii) precision measurements of the intra-protein transfer of protons and electrons under single-turnover conditions and;
(iii) theoretical modelling of the interplay between motion and charge transfer.

On studying the ATP syntheses, the intention is to reveal how does proton transfer through the membrane portion of the ATP syntheses drive the full rotation of the central shaft of this enzyme relative to the stator subunits. It is planned to correlate the effect of several parameters (site-specific mutations in Fo and F1, specific inhibitors, pH, temperature) on the active rotation of the central shaft of FoF1, as directly visualized by single molecule monitoring techniques, with the proton displacements through FoF1 in the quasi-single-enzyme mode. On studying the bc1, the intention is to clarify the coupling between the large-scale motions at the periphery of the cytochrome-bc1 complex and the electron and proton transfer in its membrane part.

Therefore it is planned to correlate the impact of various modifications of the cytochrome bc1 complex on the displacement of the peripheral FeS domain of the Rieske protein by 70° (direct visualization by new single molecule monitoring techniques would be thereby combined with indirect optical tracing via the kinetics of the cytochrome c reduction by FeS) with the proton and electron displacements inside the cytochrome-bc1 complex as monitored under the single turnover mode of the enzyme. It is planned to analyse theoretically the operation of FoF1 and bc1 by combing electrostatic calculations with the steered molecular dynamics simulations. The ultimate objective of the project is to develop mechanistic schemes of the coupling between the intramembrane charge displacements and the large-scale motions of the peripheral domains in FoF1 and in bc1.

Call for proposal

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Funding Scheme

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Coordinator

Universitat Osnabruck
EU contribution
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Address
Barbarastr. 11
49076 Osnabruck
Germany

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Total cost
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Participants (3)