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. 2011 Nov 15;6(11):e26115. doi: 10.1371/journal.pone.0026115

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Ganoth et al.

This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited.

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Everted membrane vesicles activity was determined using acridine orange fluorescence to monitor pH. (a) Data of typical measurements are shown. At the onset of the reaction, succinic acid (0.6 mM) was added to energize the vesicles and fluorescence was recorded until a steady state level of pH (100% quenching) was reached. Then, 5 mM of Na (black), Li (red), K (purple) Rb (green) or Cs (magenta) were added to evoke the Na/H antiporter. NhaA activation level was defined as the percentage of dequenching at steady state after adding Na or Li , from those 100%; (b) Michaelis-Menten kinetic fit. Activation of NhaA was measured at different Na (black) and Li (red) concentrations. Data points () are shown in circles, while the fit is shown in solid lines.

Inline graphic — Everted membrane vesicles activity was determined using acridine orange fluorescence to monitor pH. (a) Data of typical measurements are shown. At the onset of the reaction, succinic acid (0.6 mM) was added to energize the vesicles and fluorescence was recorded until a steady state level of pH (100% quenching) was reached. Then, 5 mM of Na (black), Li (red), K (purple) Rb (green) or Cs (magenta) were added to evoke the Na/H antiporter. NhaA activation level was defined as the percentage of dequenching at steady state after adding Na or Li , from those 100%; (b) Michaelis-Menten kinetic fit. Activation of NhaA was measured at different Na (black) and Li (red) concentrations. Data points () are shown in circles, while the fit is shown in solid lines.