Abstract
The catalytic free energy and binding free energies of the native and the Asn-155 .fwdarw. Thr, Asn-155 .fwdarw. Leu, and Asn-155 .fwdarw. Ala mutants of subtilisin are calculated by the empirical valence bond method and a free energy perturbation method. Two simple procedures are used; one "mutates" the substrate, and the other "mutates" the enzyme. The calculated changes in free energies between the mutant and native enzymes are within 1 kcal/mol of the corresponding observed values. This indicates that we are approaching a quantitative structure-function correlation. The calculated changes in catalytic free energies are almost entirely due to the electrostatic interaction between the enzyme-water system and the charges of the reacting system. This supports the idea that the electrostatic free enegry associated with the changes of charges of the reacting system is the key factor in enzyme catalysis.