title: How do serine proteases really work?
abstract: Recent advances in genetic engineering have led to a growing acceptance of the fact that enzymes work like other catalysts by reducing the activation barriers of the corresponding reactions. However, the key question about the action of enzymes is not related to the fact that they stabilize transition states but to the question to how they accomplish this task. This work considers the catalytic reaction of serine proteases and demonstrates how one can use a combination of calculations and experimental information to elucidate the key contributions to the catalytic free energy. Recent reports about genetic modifications of the buried aspartic group in serine proteases, which established the large effect of this group (but could not determine its origin), are analyzed. Two independent methods indicate that the buried aspartic group in serine proteases stabilizes the transition state by electrostatic interactions rather than by alternative mechanisms. Simple free energy considerations are used to eliminate the double proton-transfer mechanism (which is depicted in many textbooks as the key catalytic factor in serine proteases). The electrostatic stabilization of the oxyanion side of the transition state is also considered. It is argued that serine proteases and other enzymes work by providing electrostatic complementarity to the changes in charge distribution occurring during the reactions they catalyze.
authors.affiliation: Department of Chemistry, University of Southern California, Los Angeles 90089-0482.
authors.forename: A
authors.initials: A
authors.lastname: Warshel
authors.forename: G
authors.initials: G
authors.lastname: Naray-Szabo
authors.forename: F
authors.initials: F
authors.lastname: Sussman
authors.forename: J K
authors.initials: JK
authors.lastname: Hwang
mesh_terms: D007700:Kinetics; D008962:Models, Theoretical; D009154:Mutation; D011487:Protein Conformation; D012697:Serine Endopeptidases / Q000235:genetics / Q000378:metabolism*; D013816:Thermodynamics
chemical_list: D012697:Serine Endopeptidases