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. 1998 Dec 8;95(25):14675–14680. doi: 10.1073/pnas.95.25.14675

Table 3.

Changes in the free energies of urea-induced unfolding of p53 DNA binding domain

p53 Variant m, kcal mol−1M−1 [Urea]50%, M ΔGD−NH2O (kcal mol−1) ΔΔGD−NH2O (kcal mol−1)
WT (2) 2.40  ±  0.12 3.20  ±  0.01 8.64
Q104P 2.50  ±  0.08 3.24  ±  0.01 8.75 −0.11
Q104H 2.50  ±  0.09 3.29  ±  0.01 8.88 −0.24
A129D 3.11  ±  0.14 2.94  ±  0.01 7.94 0.70
A129E 2.83  ±  0.11 3.06  ±  0.01 8.26 0.38
A129S 3.01  ±  0.09 3.13  ±  0.01 8.45 0.19
M133L 2.60  ±  0.11 3.31  ±  0.01 8.94 −0.30
D148E 2.51  ±  0.09 3.04  ±  0.01 8.21 0.43
D148S 2.57  ±  0.09 3.28  ±  0.01 8.86 −0.22
T150P 2.86  ±  0.13 3.17  ±  0.01 8.56 0.08
Q165K 2.89  ±  0.17 2.73  ±  0.01 7.37 1.27
Q167E 2.90  ±  0.07 3.04  ±  0.06 8.21 0.43
R174K 2.80  ±  0.09 3.12  ±  0.01 8.42 0.22
C182S 2.11  ±  0.10 3.26  ±  0.02 8.80 −0.16
L201P 2.24  ±  0.09 3.33  ±  0.01 8.99 −0.35
V203A (2) 2.40  ±  0.07 3.38  ±  0.01 9.13 −0.49
L206S 2.75  ±  0.11 3.16  ±  0.01 8.55 0.10
D228E 2.40  ±  0.07 3.22  ±  0.01 8.69 −0.05
Y236F (2) 2.74  ±  0.08 3.30  ±  0.01 8.91 −0.27
N239Y 2.40  ±  0.07 3.75  ±  0.01 10.13 −1.49
S260P 2.30  ±  0.11 3.08  ±  0.01 8.32 0.32
N268D 2.97  ±  0.06 3.54  ±  0.004 9.56 −0.92
N239YN268D 3.03  ±  0.11 3.96  ±  0.01 10.68 −2.04
M133LV203A 2.50  ±  0.05 3.44  ±  0.005 9.29 −0.65
M133LN239YN268D 2.82  ±  0.10 4.03  ±  0.01 10.87 −2.23
V203AN239YN268D 2.84  ±  0.16 4.09  ±  0.01 11.05 −2.41
M133LV203AN239YN268D 2.73  ±  0.08 4.18  ±  0.01 11.29 −2.65

The mean m value of 27 denaturation curves (at 10°C in 50 mM sodium phosphate/5 mM DTT at pH 7.2) is 2.7 ± 0.05 kcal mol−1 and was used to calculate ΔGD−NH2O and ΔΔGD−NH2O according to the equation for a two-state transition as reported (11). The errors given are the standard errors, and the number of measurements is shown in parentheses. ΔGD−ND = ΔGD−NH2O − m [D] whereas ΔΔGD−N[D]50% = 〈m〉Δ[D]50%, where m is the slope of the transition from native to denatured state, [D] is the concentration of the denaturant, [D]50% is the concentration of denaturant at which 50% of the protein is unfolded, 〈m〉 is the average value of m for mutant and wild-type proteins, and Δ[D]50% = [D]50%wt − [D]50%mutant.