Charge state of His-57-Asp-102 couple in a transition state analogue-trypsin complex: a molecular orbital study

Ab initio molecular orbital studies have been made as a model for the deacylation step of trypsin. Ser-195 is modeled by H2O in which one H is replaced either by--PO2(OH)- (monoisopropyl phosphoryl, MIP) or by--CHO(OH)- (a transition state analogue, TSD). The quantum mechanical region includes imidazole+ and acetate- as models for His-57+ and Asp-102-, two hydrogen bonds from two formamide molecules to the oxyanion MIP or TSD, and three hydrogen bonds to Asp-102. The remainder of the enzyme is treated classically as a fractional charge model. The effect of proton transfer from His-57+ to Asp-102- is very similar for the MIP and TSD models, and the proton transfer is energetically unfavorable for all models that include at least the hydrogen bond from an H2O that models Ser-214. Thus, the several hydrogen bonds to the models of the catalytic unit (substrate, Ser-195, His-57, and Asp-102) stabilize the His-57+/Asp-102- salt link, and this indicates that proton transfer does not occur from His-57+ to Asp-102-. (Also, the similarities of energy of transfer of this proton transfer for the various models show that the model substrate analogue behaves very similarly to the MIP inhibitor.)