@inproceedings{67130eba201c4685b6b0620e23d65404,
title = "Stepwise rotation of the γ-subunit of EFoF1-ATP synthase during ATP synthesis: a single-molecule FRET approach",
abstract = "FoF1-ATP synthases couple proton translocation with the synthesis of ATP using two rotary motors within the enzyme. To monitor inter-subunit movements during catalysis, we selectively attached two fluorophores to the F 1 part, sulforhodamine B at one of three {\ss}-subunits and Cy5 at the ?-subunit. Reassembly with F o parts embedded in liposomes yielded functional holoenzymes. Fluorescence resonance energy transfer (FRET) was investigated in photon bursts of freely diffusing liposomes with reconstituted ATP synthases using a confocal set-up for single-molecule detection. Incubation with AMPPNP resulted in stable intensity ratios within a burst and three different FRET efficiencies. Upon ATP addition, a repeating sequence of three distinct FRET efficiencies was observed, indicating the stepwise movement of the ?-subunit during ATP hydrolysis. With this single-molecule FRET approach we detected a stepwise rotation of the ?-subunit under conditions for ATP synthesis (i.e. energization of the proteoliposomes by an acid-base-transition). The direction of rotation is opposite to the direction observed during ATP hydrolysis.",
author = "Michael Borsch and Manuel Diez and Boris Zimmermann and Matthias Trost and Stefan Steigmiller and Peter Graber",
year = "2003",
doi = "10.1117/12.479554",
language = "English",
isbn = "0819447625",
series = "Progress in Biomedical Optics",
publisher = "SPIE-International Society for Optical Engineering",
number = "14",
pages = "11--21",
editor = "Nicolau, {Dan V.} and Joerg Enderlein and Leif, {Robert C.} and Farkas, {Daniel L.}",
booktitle = "Manipulation and Analysis of Biomolecules, Cells, and Tissues",
note = "SPIE Photonics West 2003: Manipulation and Analysis of Biomolecules, Cells and Tissues ; Conference date: 28-01-2003 Through 29-01-2003",
}