Table 1.
ΔG (forward) (kJ/mol) | ΔG (backward) (kJ/mol) | ΔG (average) (kJ/mol) | ΔΔG (kJ/mol) | |
---|---|---|---|---|
Tripeptide | ||||
L78A | 1.7 | 5.7 | 3.7±2.0 | |
L81A | 3.2 | 4.0 | 3.6±0.4 | |
M202A | 7.1 | 6.3 | 6.7±0.4 | |
Protein | ||||
M202A | 23.3 | 11.5 | 17.4±5.9 | 10.8±6.3 |
L81A/M202A | 46.7 | 31.3 | 39.0±7.7 | 28.7±8.5 |
L78A/L81A/M202A | 66.2 | 52.6 | 59.4±6.8 | 45.4±9.6 |
Free energy changes, ΔG, between WT and mutants for the unfolded and folded state, respectively, and differences, ΔΔG, as an estimate of the effect of the mutation on the thermodynamic stability of the complex. An estimate for the uncertainty is obtained by comparing forward and backward ΔG values. The single, double and triple mutants display a progressive amount of destabilization with respect to the WT protein. |