Table 1.
Calculated and observed Δg‡cat for the reaction of CALA and its mutants. [a]
| mutations [b] | label | enantiomer | ΔG0calc. (PT), kcal/mol | Δg‡calc. (NA), kcal/mol | Δg‡calc. (total), kcal/mol | Δg‡exp. (total), kcal/mol | E′exp. [c] | E′calc. [c] |
|---|---|---|---|---|---|---|---|---|
| water | 1 | R/S | 11.9 | 11.3 | 23.2 | 23.3 | - | - |
| wild type | 2 | R | 13.1 | 5.2 | 18.3 | 17.9 | 3.8 (S) | 1.7 (S) |
| S | 14.5 | 3.5 | 18.0 | 17.1 | ||||
| F233L/G237Y | 3 | R | 12.1 | 4.7 | 16.8 | 16.9 | 7.6 (R) | 6.4 (R) |
| S | 14.1 | 3.8 | 17.9 | 18.1 | ||||
| T64M/F149S/I150D/F233N/G237L | 4 | R | 13.1 | 4.5 | 17.6 | 18.6 | 11.0 (S) | 5.4 (S) |
| S | 14.2 | 2.4 | 16.6 | 17.2 | ||||
| T64M/F149S/I150D/Y183F/F233N/G237L | 5 | R | 13.8 | 4.6 | 18.4 | - | no exp. | 14.9 (S) |
| S | 14.5 | 2.3 | 16.8 | - | ||||
| Y183F | 6 | R | 15.2 | 3.2 | 18.4 | - | no exp. | 6.4 (S) |
| S | 12.5 | 4.8 | 17.3 | - | ||||
| G237Y | 7 | R | 11.4 | 5.1 | 16.5 | - | no exp. | 6.4 (S) |
| S | 13.1 | 2.3 | 15.4 | - | ||||
| F233N | 8 | R | 13.2 | 5.2 | 18.4 | - | no exp. | 24.8 (S) |
| S | 13.3 | 3.2 | 16.5 | - | ||||
| F233G | 9 | R | 10.6 | 3.9 | 14.5 | - | Eexp.= 17 (R) | 12.6 (R) |
| S | 13.2 | 2.8 | 16.0 | - | ||||
| F149Y/I150N/F233G | 10 | R | 12.6 | 2.9 | 15.5 | - | Eexp.= 104 (R) | 68.5 (R) |
| S | 14.3 | 3.7 | 18.0 | - | ||||
| F233L | 11 | R | 11.5 | 5.6 | 17.1 | - | no exp. | 3.3 (R) |
| S | 14.0 | 3.8 | 17.8 | - |
[a]
The calculated Δg‡cat (in kcal/mol) reflects an average over 10 conformations obtained from equally spaced points along the relaxation trajectory. The standard deviation is reported in Table S3.
[b]
The X-ray structure 2VEO.pdb of the wild type CALA [27] were used as the initial geometry for the subsequent wt relaxation and EVB calculations. The initial structures for the different mutations were generated from the wt CALA X-ray structure, using the PyMOL molecular graphics software, [33] following by 200ps relaxation run.