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. 1999 Jul 6;96(14):7780–7784. doi: 10.1073/pnas.96.14.7780

Figure 3.

Figure 3

ATP-dependent formation of an electrochemical proton gradient in membrane vesicles containing the engineered FOF1 with or without the γM23K mutation. ATP-dependent fluorescence quenching in membrane vesicles containing the engineered enzyme with (dotted line) or without (solid line) the γM23K mutation. Membrane vesicles (100 μg of protein) were suspended in the reaction mixture for the rotation assay containing 2 μM acridine orange. Fluorescence (F) at 530 nm was monitored at 25°C. At the time indicated (arrowhead), 10 μl of 100 mM ATP (1 mM final concentration) or 10 μl of 1 M NH4Cl (10 mM) was added. Essentially the same results were obtained with 6 mM ATP and 2 μM quinacrine. Note that measurements were carried out under the conditions for the rotation assay.