Figure 7. Nonadditive effects of mutations on the ATP modulations of CLC-1. The parameter Poc(ATP)/Poc(control) was used as the response to ATP modulation. (A) Mutant cycle analysis of the effects of V634F and V613A mutations on the ATP modulation. ATP dose–response curves of single point mutants and the double mutants were constructed to determine the values of Ko and Kc. The fitted Kc values were used to calculate the free energy differences of single or double mutants (bar graph) according to Eq. 3 and the coupling constant, Ω, according to Eq. 4. In the context of Eq. 2, the V634F mutation corresponds to the change from X1 to X2, whereas the V613A mutation represents the change from Y1 to Y2. (B) Double mutant cycle analysis for V634F/E865D mutations. The values of Ω at different voltages are shown in Fig. S2. Error bars represent SEM. n = 4–8.
Figure 7.

Nonadditive effects of mutations on the ATP modulations of CLC-1. The parameter Poc(ATP)/Poc(control) was used as the response to ATP modulation. (A) Mutant cycle analysis of the effects of V634F and V613A mutations on the ATP modulation. ATP dose–response curves of single point mutants and the double mutants were constructed to determine the values of Ko and Kc. The fitted Kc values were used to calculate the free energy differences of single or double mutants (bar graph) according to Eq. 3 and the coupling constant, Ω, according to Eq. 4. In the context of Eq. 2, the V634F mutation corresponds to the change from X1 to X2, whereas the V613A mutation represents the change from Y1 to Y2. (B) Double mutant cycle analysis for V634F/E865D mutations. The values of Ω at different voltages are shown in Fig. S2. Error bars represent SEM. n = 4–8.

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