Table 1.
Protein region | Mutant | ΔΔGD-N, kcal· mol−1 | ΦTS1 | ΦTS2 |
---|---|---|---|---|
Mutation compared with wild type | ||||
β1 | I314V | −0.44 ± 0.08* | ||
β1 | I316A | 3.5 ± 0.2 | 0.28 ± 0.04 | 0.52 ± 0.03 |
Loop | L323A | 1.87 ± 0.08 | −0.20 ± 0.03 | 0.38 ± 0.05 |
β2 | F325A | 4.9 ± 1.0 | 0.36 ± 0.13 | 0.67 ± 0.07 |
β2 | I327V | 0.96 ± 0.07 | 0.25 ± 0.03 | 0.52 ± 0.09 |
β2 | V328A | 0.32 ± 0.08* | ||
β3 | I336V | 0.45 ± 0.07* | ||
β3 | I336A | 1.35 ± 0.08 | 0.01 ± 0.03 | 0.53 ± 0.06 |
β3 | I338A | 2.2 ± 0.1 | 0.15 ± 0.03 | 0.53 ± 0.04 |
β3 | F340A | 0.12 ± 0.08* | ||
β3 | I341V | −0.16 ± 0.08* | ||
β3 | I341A | 1.57 ± 0.08 | −0.08 ± 0.03 | 0.42 ± 0.05 |
α1 | P346G | 1.33 ± 0.07 | −0.05 ± 0.03 | 0.27 ± 0.07 |
α1 | A347G | 1.32 ± 0.07 | −0.05 ± 0.03 | 0.20 ± 0.07 |
Loop | L353A | 2.24 ± 0.09 | 0.15 ± 0.02 | 0.50 ± 0.04 |
β4 | I359V | 0.94 ± 0.07 | 0.36 ± 0.04 | 0.60 ± 0.09 |
β4 | V362A | 1.68 ± 0.08 | 0.60 ± 0.02 | 0.87 ± 0.06 |
α2 | H372A | −1.3 ± 0.1 | 0.87 ± 0.05 | 0.77 ± 0.09 |
α2 | E373A | 0.05 ± 0.08* | ||
α2 | E373G | 0.65 ± 0.08 | 0.50 ± 0.06 | 0.62 ± 0.14 |
α2 | A375G | 0.98 ± 0.07 | 0.79 ± 0.05 | 1.04 ± 0.13 |
α2 | A376G | 1.35 ± 0.07 | 0.55 ± 0.03 | 0.84 ± 0.08 |
α2 | I377A | 0.00 ± 0.08* | ||
α2 | I377G | 1.21 ± 0.07 | 0.58 ± 0.03 | 0.75 ± 0.08 |
α2 | A378G | 2.4 ± 0.1 | 0.34 ± 0.02 | 0.60 ± 0.04 |
α2 | L379A | 1.52 ± 0.07 | 0.59 ± 0.03 | 0.82 ± 0.07 |
α2 | K380A | −0.58 ± 0.08 | 0.77 ± 0.09 | 0.69 ± 0.16 |
α2 | K380G | 0.53 ± 0.07 | 0.18 ± 0.07 | 0.75 ± 0.19 |
Turn | A382G | 1.19 ± 0.08 | 0.38 ± 0.03 | 0.59 ± 0.07 |
β6 | V386A | 2.00 ± 0.07 | 0.16 ± 0.02 | 0.56 ± 0.04 |
β6 | I388V | 1.44 ± 0.07 | 0.35 ± 0.02 | 0.56 ± 0.06 |
β6 | Y392A | 1.51 ± 0.08 | 0.17 ± 0.03 | 0.96 ± 0.08 |
Val→Ala or Ala→Gly | ||||
β3 | V336A | 0.90 ± 0.08 | 0.00 ± 0.05 | 0.33 ± 0.10 |
β3 | V341A | 1.73 ± 0.08 | −0.03 ± 0.03 | 0.45 ± 0.05 |
α2 | A373G | 0.60 ± 0.08 | 0.42 ± 0.06 | 0.51 ± 0.14 |
α2 | A377G | 1.21 ± 0.08 | 0.66 ± 0.04 | 0.78 ± 0.08 |
α2 | A380G | 1.11 ± 0.08 | 0.49 ± 0.03 | 0.72 ± 0.09 |
The standard errors of curve fitting for the kinetic parameters were similar to standard deviations, as determined from three independent kinetic measurements on the pseudo-wild-type PDZ3 (Fig. 2A). Therefore, fitting errors of kf, ku, and Kp were used to calculate the propagated errors in Φ and Δ Δ GD-N as described (50).
*The ΔΔ GD-N value is too low to calculate an accurate Φ value.