Table 1.

Relative apparent free energies of channel activation at hyperpolarized and depolarized holding potentials

Channel V1/2 (0 mV) Z (0 mV) ΔG (0 mV) V1/2 (−80 mV) Z (−80 mV) ΔG (−80 mV) ΔΔG 
 mV  kcal/mol mV  kcal/mol kcal/mol 
WT −127.8 ± 5.6 −1.9 ± 0.1 −5.4 ± 0.2 −86.9 ± 1.7 −1.6 ± 0.0 −3.3 ± 0.1 2.10 
R1C −108.8 ± 6.5 −1.1 ± 0.1 −2.7 ± 0.1 −102.3 ± 6.9 −0.9 ± 0.0 −2.4 ± 0.1 0.38 
R2C −115.7 ± 3.0 −1.0 ± 0.0 −2.9 ± 0.1 −95.2 ± 0.4 −1.3 ± 0.2 −3.0 ± 0.2 0.02 
R3C −129.6 ± 3.0 −1.6 ± 0.0 −4.9 ± 0.2 −105.3 ± 0.2 −1.4 ± 0.2 −3.3 ± 0.4 1.62 
Channel V1/2 (0 mV) Z (0 mV) ΔG (0 mV) V1/2 (−80 mV) Z (−80 mV) ΔG (−80 mV) ΔΔG 
 mV  kcal/mol mV  kcal/mol kcal/mol 
WT −127.8 ± 5.6 −1.9 ± 0.1 −5.4 ± 0.2 −86.9 ± 1.7 −1.6 ± 0.0 −3.3 ± 0.1 2.10 
R1C −108.8 ± 6.5 −1.1 ± 0.1 −2.7 ± 0.1 −102.3 ± 6.9 −0.9 ± 0.0 −2.4 ± 0.1 0.38 
R2C −115.7 ± 3.0 −1.0 ± 0.0 −2.9 ± 0.1 −95.2 ± 0.4 −1.3 ± 0.2 −3.0 ± 0.2 0.02 
R3C −129.6 ± 3.0 −1.6 ± 0.0 −4.9 ± 0.2 −105.3 ± 0.2 −1.4 ± 0.2 −3.3 ± 0.4 1.62 

Apparent free energy for a two-state model, ΔG, was calculated as a measure for comparing the relative energies of opening WT and mutant channels at holding potentials of 0 or −80 mV as ΔG = −ZFV1/2. Z was the apparent charge, V1/2 the half-activation voltage of charge movement, and F, Faraday’s number (23.061 kcal per volt gram equivalent). The parameters V1/2 and Z were calculated by using the slopes of fits of 1/(1 + exp[RT(V − V1/2)/ZF] to Q-V curves, where R is the universal gas constant, T is temperature in °K, F is the Faraday constant, and Z is the valence. ΔΔG was calculated as ΔΔG = ΔG (−80 mV) − ΔG (0 mV).

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