TABLE 2.
Thermodynamic and kinetic data for mutants of TNfn3
| Monomer
|
8-mer
|
||||
|---|---|---|---|---|---|
| Mutation (strand) | [Urea]50% (M)* | ΔΔGD − N* kcal mol−1 | [urea]50% (M) | ΔΔGD − N† kcal mol−1 | kf (s−1)‡ |
| Wild-type | 5.33 (0.03) | — | 6.00 (0.05) | — | 1.20 (0.03) |
| L2A (A) | 3.59 (0.1) | 2.1 (0.1) | 4.35 (0.07) | 2.0 (0.1) | 1.43 (0.13) |
| I8A (A) | 2.98 (0.1) | 2.9 (0.1) | 3.99 (0.06) | 2.5 (0.1) | 1.34 (0.11) |
| I20A (B) | 2.31 (0.1) | 3.7 (0.1) | 3.48 (0.08) | 3.1 (0.1) | 0.20 (0.01) |
| Y36A (C) | 1.86 (0.1) | 4.2 (0.1) | 2.86 (0.03) | 3.8 (0.1) | 0.07 (0.01) |
| I48A (C′) | 3.52 (0.1) | 2.2 (0.1) | 4.35 (0.09) | 2.0 (0.1) | 0.13 (0.01) |
| I59A (E) | 3.63 (0.1) | 2.1 (0.1) | 4.30 (0.07) | 2.1 (0.1) | 0.26 (0.03) |
| Y68F (F) | 2.75 (0.1) | 3.1 (0.1) | 3.53 (0.03) | 3.0 (0.1) | 0.04 (0.01) |
| V70A (F) | 3.01 (0.1) | 2.8 (0.1) | 3.73 (0.05) | 2.8 (0.1) | 0.25 (0.01) |
| T90A (G) | 2.50 (0.1) | 3.4 (0.1) | 3.52 (0.10) | 3.0 (0.1) | 0.37 (0.01) |
*
Values previously reported in Cota et al. (2000).
†
The change in ΔGD − N on mutation. Calculated using a mean m-value of 1.22 (0.05) kcal mol−1 M−1 derived from the studies of the monomer (Hamill et al., 2000).
‡
kf at 0 M denaturant at pH 5.8 as determined by pH-jump.