Quantum mechanical study on the catalytic mechanism of alkaline phosphatases

BOROSKY, GABRIELA L.

JOURNAL OF CHEMICAL INFORMATION AND MODELING

AMER CHEMICAL SOC

Lugar: Washington; Año: 2017

1549-9596

Alkaline phosphatases (APs) catalyze the hydrolysis and transphosphorylation of phosphate monoesters. The catalytic mechanism was examined by quantum-mechanical calculations, using an active-site model based on the X-ray crystal structure of the human placental AP (PLAP). Free energies of activation and of reaction of the catalytic steps were evaluated for a series of aryl and alkyl phosphate esters, and computational results were compared with experimental values available in the literature. Mechanistical observations previously reported in experimental works were rationalized by the present theoretical study, particularly regarding the difference in the rate-determining step between aryl and alkyl phosphates. The formation rate of the covalent phosphoserine intermediate followed a linear free-energy relationship (LFER) with the pKa of the leaving group. This LFER, that could only be experimentally determined for less reactive alkyl phosphates, was verified by the present calculations to apply for the entire set of aryl and alkyl phosphate substrates.