Table II.

Summary of experimental data from our mutagenesis screen

 pos V1/2 a ΔV1/2 ΔΔGa VDI50 VDI300 CDI50 CDI300 Figure 
WT  −26.8 1 0.0 0.00 0.022 ± 0.006 0.091 ± 0.032 0.75 ± 0.03 0.80 ± 0.03 4 A (i) 
G0384P −8 −10.1 0.18 16.7 1.03 0.029 ± 0.006 0.081 ± 0.033 0.59 ± 0.04 0.73 ± 0.03 4 A (i) 
L0393P 1 −6.7 0.12 20.1 1.27 0.021 ± 0.005 0.054 ± 0.014 0.26 ± 0.03 0.50 ± 0.03 4 B (i) 
G0394P 2 6.5 0.045 33.4 1.86 0.033 ± 0.011 0.044 ± 0.013 0.06 ± 0.03 0.10 ± 0.03 4 B (i) 
V0395P 3 −7.9 0.07 19.0 1.60 0.042 ± 0.015 0.147 ± 0.053 0.76 ± 0.03 0.84 ± 0.01 4 A (ii) 
L0396P 4 −33.2 2.5 −6.3 −0.55 0.054 ± 0.012 0.207 ± 0.038 0.78 ± 0.02 0.85 ± 0.02 4 A (ii) 
S0397P 5 −42.0 10 −15.2 −1.38 0.003 ± 0.001 −0.008 ± 0.008 0.30 ± 0.05 0.33 ± 0.06 4 A (i) 
G0398P 6 −16.2 0.25 10.6 0.83 0.011 ± 0.008 0.064 ± 0.044 0.75 ± 0.03 0.83 ± 0.02 4 A (i) 
E0399P 7 −25.3 0.8 1.5 0.13 0.117 ± 0.020 0.402 ± 0.025 0.79 ± 0.01 0.85 ± 0.01 4 A (ii) 
F0400P 8 −20.9 0.5 6.0 0.42 0.159 ± 0.015 0.519 ± 0.035 0.72 ± 0.03 0.84 ± 0.03 4 A (ii) 
L0759P 1 −42.8 14 −15.9 −1.58 0.004 ± 0.003 −0.051 ± 0.021 0.10 ± 0.01 0.18 ± 0.02 4 A (i) 
A0760P 2 −42.3 12 −15.5 −1.49 0.005 ± 0.001 0.032 ± 0.015 0.26 ± 0.02 0.48 ± 0.01 4 A (i) 
I0761P 3 −64.2 200 −37.4 −3.18 0.008 ± 0.001 0.048 ± 0.007 0.14 ± 0.06 0.21 ± 0.07 4 A (i) 
A0762P 4 −47.7 15 −20.9 −1.62 0.011 ± 0.002 −0.001 ± 0.023 0.17 ± 0.06 0.22 ± 0.06 4 A (i) 
V0763P 5 −57.1 20 −30.3 −1.80 0.008 ± 0.002 0.027 ± 0.008 0.31 ± 0.03 0.40 ± 0.03 4 A (i) 
D0764P 6 −36.8 5 −10.0 −0.97 0.019 ± 0.011 0.085 ± 0.033 0.26 ± 0.03 0.44 ± 0.04 4 A (i) 
N0765P 7 −44.5 11 −17.6 −1.44 0.014 ± 0.006 0.089 ± 0.028 0.42 ± 0.01 0.56 ± 0.01 4 A (i) 
L0766P 8 −40.6 6 −13.7 −1.08 0.022 ± 0.005 0.097 ± 0.024 0.51 ± 0.08 0.57 ± 0.07 4 A (i) 
V1159P 1 −31.2 2 −4.4 −0.42 0.010 ± 0.002 0.032 ± 0.009 0.54 ± 0.01 0.58 ± 0.01 4 A (i) 
G1160P 2 −47.5 19 −20.7 −1.77 0.014 ± 0.003 0.045 ± 0.011 0.34 ± 0.04 0.55 ± 0.05 4 A (i) 
F1161P 3 −45.9 21 −19.1 −1.83 0.020 ± 0.008 0.148 ± 0.012 0.30 ± 0.04 0.37 ± 0.04 4 A (i) 
V1162P 4 −38.8 5 −12.0 −0.97 0.068 ± 0.020 0.260 ± 0.036 0.58 ± 0.05 0.83 ± 0.04 4 A (iii) 
I1163P 5 −46.1 23 −19.3 −1.88 0.035 ± 0.008 0.119 ± 0.015 0.11 ± 0.02 0.30 ± 0.03 4 A (i) 
V1164P 6 −50.7 48 −23.8 −2.32 0.016 ± 0.007 0.070 ± 0.011 0.20 ± 0.05 0.33 ± 0.04 4 A (i) 
T1165P 7 −51.7 100 −24.9 −2.76 0.021 ± 0.014 0.058 ± 0.011 0.27 ± 0.04 0.60 ± 0.06 4 A (i) 
F1166P 8 −37.5 4.5 −10.7 −0.90 0.120 ± 0.027 0.490 ± 0.067 0.69 ± 0.03 0.89 ± 0.01 4 A (ii) 
V1456P 1 −13.2 0.12 13.7 1.27 0.032 ± 0.008 0.111 ± 0.037 0.71 ± 0.02 0.73 ± 0.01 4 A (i) 
A1457P 2 −23.8 0.7 3.0 0.21 0.039 ± 0.010 0.179 ± 0.021 0.64 ± 0.01 0.76 ± 0.01 4 A (iii) 
V1458P 3 −17.2 0.35 9.6 0.63 0.029 ± 0.013 0.112 ± 0.031 0.65 ± 0.02 0.74 ± 0.02 4 A (i) 
I1459P 4 −15.9 0.25 10.9 0.83 0.011 ± 0.003 0.060 ± 0.031 0.77 ± 0.02 0.80 ± 0.02 4 A (i) 
M1460P 5 −13.4 0.22 13.4 0.91 0.012 ± 0.002 0.027 ± 0.045 0.41 ± 0.02 0.58 ± 0.01 4 B (i) 
D1461P 6 −25.2 0.85 1.6 0.10 0.038 ± 0.006 0.130 ± 0.014 0.67 ± 0.03 0.74 ± 0.02 4 A (i) 
N1462P 7 −31.4 2 −4.5 −0.42 0.009 ± 0.002 0.042 ± 0.016 0.53 ± 0.02 0.55 ± 0.02 4 A (i) 
F1463P 8 −2.2 0.06 24.7 1.69 0.256 ± 0.014 0.391 ± 0.027 0.22 ± 0.02 0.38 ± 0.03 4 B (iii) 
D1464P 9 −7.7 0.09 19.1 1.44 0.125 ± 0.019 0.464 ± 0.031 0.68 ± 0.03 0.75 ± 0.05 4 A (ii) 
WT/BayK  −39.2 6.5 −12.4 −1.12 0.022 ± 0.006 0.140 ± 0.038 0.46 ± 0.06 0.62 ± 0.03 5 B  
V0395P/BayK 3 −14.1 0.15 12.7 1.14 0.007 ± 0.003 0.106 ± 0.032 0.78 ± 0.02 0.80 ± 0.02 5 B  
N0765P/BayK 7 −56.0 125 −29.2 −2.90 0.010 ± 0.004 0.021 ± 0.011 0.12 ± 0.01 0.31 ± 0.03 5 B  
V1162P/BayK 4 −37.4 5 −10.6 −0.97 0.022 ± 0.008 0.183 ± 0.046 0.62 ± 0.03 0.81 ± 0.03 5 B  
 pos V1/2 a ΔV1/2 ΔΔGa VDI50 VDI300 CDI50 CDI300 Figure 
WT  −26.8 1 0.0 0.00 0.022 ± 0.006 0.091 ± 0.032 0.75 ± 0.03 0.80 ± 0.03 4 A (i) 
G0384P −8 −10.1 0.18 16.7 1.03 0.029 ± 0.006 0.081 ± 0.033 0.59 ± 0.04 0.73 ± 0.03 4 A (i) 
L0393P 1 −6.7 0.12 20.1 1.27 0.021 ± 0.005 0.054 ± 0.014 0.26 ± 0.03 0.50 ± 0.03 4 B (i) 
G0394P 2 6.5 0.045 33.4 1.86 0.033 ± 0.011 0.044 ± 0.013 0.06 ± 0.03 0.10 ± 0.03 4 B (i) 
V0395P 3 −7.9 0.07 19.0 1.60 0.042 ± 0.015 0.147 ± 0.053 0.76 ± 0.03 0.84 ± 0.01 4 A (ii) 
L0396P 4 −33.2 2.5 −6.3 −0.55 0.054 ± 0.012 0.207 ± 0.038 0.78 ± 0.02 0.85 ± 0.02 4 A (ii) 
S0397P 5 −42.0 10 −15.2 −1.38 0.003 ± 0.001 −0.008 ± 0.008 0.30 ± 0.05 0.33 ± 0.06 4 A (i) 
G0398P 6 −16.2 0.25 10.6 0.83 0.011 ± 0.008 0.064 ± 0.044 0.75 ± 0.03 0.83 ± 0.02 4 A (i) 
E0399P 7 −25.3 0.8 1.5 0.13 0.117 ± 0.020 0.402 ± 0.025 0.79 ± 0.01 0.85 ± 0.01 4 A (ii) 
F0400P 8 −20.9 0.5 6.0 0.42 0.159 ± 0.015 0.519 ± 0.035 0.72 ± 0.03 0.84 ± 0.03 4 A (ii) 
L0759P 1 −42.8 14 −15.9 −1.58 0.004 ± 0.003 −0.051 ± 0.021 0.10 ± 0.01 0.18 ± 0.02 4 A (i) 
A0760P 2 −42.3 12 −15.5 −1.49 0.005 ± 0.001 0.032 ± 0.015 0.26 ± 0.02 0.48 ± 0.01 4 A (i) 
I0761P 3 −64.2 200 −37.4 −3.18 0.008 ± 0.001 0.048 ± 0.007 0.14 ± 0.06 0.21 ± 0.07 4 A (i) 
A0762P 4 −47.7 15 −20.9 −1.62 0.011 ± 0.002 −0.001 ± 0.023 0.17 ± 0.06 0.22 ± 0.06 4 A (i) 
V0763P 5 −57.1 20 −30.3 −1.80 0.008 ± 0.002 0.027 ± 0.008 0.31 ± 0.03 0.40 ± 0.03 4 A (i) 
D0764P 6 −36.8 5 −10.0 −0.97 0.019 ± 0.011 0.085 ± 0.033 0.26 ± 0.03 0.44 ± 0.04 4 A (i) 
N0765P 7 −44.5 11 −17.6 −1.44 0.014 ± 0.006 0.089 ± 0.028 0.42 ± 0.01 0.56 ± 0.01 4 A (i) 
L0766P 8 −40.6 6 −13.7 −1.08 0.022 ± 0.005 0.097 ± 0.024 0.51 ± 0.08 0.57 ± 0.07 4 A (i) 
V1159P 1 −31.2 2 −4.4 −0.42 0.010 ± 0.002 0.032 ± 0.009 0.54 ± 0.01 0.58 ± 0.01 4 A (i) 
G1160P 2 −47.5 19 −20.7 −1.77 0.014 ± 0.003 0.045 ± 0.011 0.34 ± 0.04 0.55 ± 0.05 4 A (i) 
F1161P 3 −45.9 21 −19.1 −1.83 0.020 ± 0.008 0.148 ± 0.012 0.30 ± 0.04 0.37 ± 0.04 4 A (i) 
V1162P 4 −38.8 5 −12.0 −0.97 0.068 ± 0.020 0.260 ± 0.036 0.58 ± 0.05 0.83 ± 0.04 4 A (iii) 
I1163P 5 −46.1 23 −19.3 −1.88 0.035 ± 0.008 0.119 ± 0.015 0.11 ± 0.02 0.30 ± 0.03 4 A (i) 
V1164P 6 −50.7 48 −23.8 −2.32 0.016 ± 0.007 0.070 ± 0.011 0.20 ± 0.05 0.33 ± 0.04 4 A (i) 
T1165P 7 −51.7 100 −24.9 −2.76 0.021 ± 0.014 0.058 ± 0.011 0.27 ± 0.04 0.60 ± 0.06 4 A (i) 
F1166P 8 −37.5 4.5 −10.7 −0.90 0.120 ± 0.027 0.490 ± 0.067 0.69 ± 0.03 0.89 ± 0.01 4 A (ii) 
V1456P 1 −13.2 0.12 13.7 1.27 0.032 ± 0.008 0.111 ± 0.037 0.71 ± 0.02 0.73 ± 0.01 4 A (i) 
A1457P 2 −23.8 0.7 3.0 0.21 0.039 ± 0.010 0.179 ± 0.021 0.64 ± 0.01 0.76 ± 0.01 4 A (iii) 
V1458P 3 −17.2 0.35 9.6 0.63 0.029 ± 0.013 0.112 ± 0.031 0.65 ± 0.02 0.74 ± 0.02 4 A (i) 
I1459P 4 −15.9 0.25 10.9 0.83 0.011 ± 0.003 0.060 ± 0.031 0.77 ± 0.02 0.80 ± 0.02 4 A (i) 
M1460P 5 −13.4 0.22 13.4 0.91 0.012 ± 0.002 0.027 ± 0.045 0.41 ± 0.02 0.58 ± 0.01 4 B (i) 
D1461P 6 −25.2 0.85 1.6 0.10 0.038 ± 0.006 0.130 ± 0.014 0.67 ± 0.03 0.74 ± 0.02 4 A (i) 
N1462P 7 −31.4 2 −4.5 −0.42 0.009 ± 0.002 0.042 ± 0.016 0.53 ± 0.02 0.55 ± 0.02 4 A (i) 
F1463P 8 −2.2 0.06 24.7 1.69 0.256 ± 0.014 0.391 ± 0.027 0.22 ± 0.02 0.38 ± 0.03 4 B (iii) 
D1464P 9 −7.7 0.09 19.1 1.44 0.125 ± 0.019 0.464 ± 0.031 0.68 ± 0.03 0.75 ± 0.05 4 A (ii) 
WT/BayK  −39.2 6.5 −12.4 −1.12 0.022 ± 0.006 0.140 ± 0.038 0.46 ± 0.06 0.62 ± 0.03 5 B  
V0395P/BayK 3 −14.1 0.15 12.7 1.14 0.007 ± 0.003 0.106 ± 0.032 0.78 ± 0.02 0.80 ± 0.02 5 B  
N0765P/BayK 7 −56.0 125 −29.2 −2.90 0.010 ± 0.004 0.021 ± 0.011 0.12 ± 0.01 0.31 ± 0.03 5 B  
V1162P/BayK 4 −37.4 5 −10.6 −0.97 0.022 ± 0.008 0.183 ± 0.046 0.62 ± 0.03 0.81 ± 0.03 5 B  

Each mutant is specified by name (first column) and position (second column, corresponding to S6 coordinate in Fig. 1 A). Parameters are as defined in Table I. VDI and CDI metrics are mean ± SEM. The final column indicates the figure in which the data are plotted (Figs. 4 A, 4 B, or 5 B), followed by the corresponding zone (i, ii, or iii) in Fig. 5 C.

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