Open AccessToward computer-aided site-directed mutagenesis of enzymes.
Author(s) -
Arieh Warshel,
Fredy Sussman
Publication year - 1986
Publication title -
proceedings of the national academy of sciences of the united states of america
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 5.011
H-Index - 771
eISSN - 1091-6490
pISSN - 0027-8424
DOI - 10.1073/pnas.83.11.3806
Subject(s) - mutagenesis , limiting , enzyme , site directed mutagenesis , chemistry , trypsin , ionic bonding , active site , hydrolysis , biochemistry , combinatorial chemistry , biophysics , biology , mutation , organic chemistry , mutant , engineering , ion , mechanical engineering , gene
A preliminary attempt to simulate the observed effect of a site-directed mutagenesis of rat trypsin gives encouraging results. The calculations reproduce in a semiquantitative way the observed change in the activation barrier of the rate-limiting step of amide hydrolysis. This result, which did not require any adjustable parameters, indicates that our method may provide a reliable basis for computer-aided enzyme design. In addition to the potentially practical value of the calculations, they provide important mechanistic information--that is, the change in the catalytic effect in trypsin appears to be almost exclusively due to the change in the electrostatic stabilization of the ionic configurations. This supports the view that electrostatic effects are the major factor in enzyme catalysis.