Figure 3
From: The lateral distance between a proton pump and ATP synthase determines the ATP-synthesis rate
Auto-correlation data measured with GUVs containing co-reconstituted cyt. bo 3 and ATP synthase. GUVs were composed of either 99% DOPC (A–C) or 94% DOPC and 5% DOPG (D–F), with the addition of 1% DPPE functionalized with a biotinyl head group. Measurements were done at pH 7.4 in 10 mM HEPES supplemented with 10 mM NaCl and 100 mM glucose. (A,D) FCS was used to study samples where cyt. bo 3 was labeled with either ATTO 647N (red trace) or ATTO 594 (green trace). (B,E) samples with cyt. bo 3 labeled with ATTO 647N and ATP-synthase labeled with ATTO 594. The dashed lines represent best fits of the data using a single component with planar two-dimensional diffusion and a triplet state fraction. The amplitude of the diffusional component obtained from the fit of the data with the ATTO 594-labeled protein has been set to unity (amplitude at ~10−4 s) to facilitate comparison of the traces. The autocorrelation function for FCCS was calculated in all cases and the normalized cross correlation amplitudes are compared in panels (C) and (F).
