Theoretical Study of VX Hydrolysis Mechanism Catalyzed by Phosphotriesterase Mutant H254R

Phosphotriesterase (PTE) is a bacterial enzyme that can catalyze the hydrolysis of V-type chemical warfare nerve agents. Herein, the hydrolysis mechanism of VX analogs catalyzed by the H254R PTE mutant, in which the histidine 254 residue is replaced by an arginine, has been investigated using the hybrid density functional theory method B3LYP. The chemical model of the active site of H254R PTE mutant was constructed based on the high resolution X-ray crystal structure. We have presented the energetics and the characterization of stationary points in the catalytic reaction. The calculations indicate that the catalytic process contain two SN2 displacement steps and the step of hydroxide regeneration is rate-limiting with an overall barrier of 22.2 kcal/mol. The Arg254 plays an important role as a proton donor/acceptor for protonating leaving group and activating water molecule. Moreover, the H254R PTE mutant with binuclear cobalt center turns out to higher activity towards VX than the binuclear zinc H254R PTE.