Dineopentyl phosphate hydrolysis: evidence for stepwise water attack

J Org Chem. 2008 Sep 19;73(18):6960-9. doi: 10.1021/jo801207q. Epub 2008 Aug 27.

Abstract

Phosphate ester hydrolysis is ubiquitous in biology, playing a central role in energy production, signaling, biosynthesis, and the regulation of protein function among other things. Although the mechanism of action of the enzymes regulating this reaction has been the focus of intensive research in the past few decades, the correct description of this apparently simple reaction remains controversial. A clear understanding of the mechanism that takes place in solution is crucial to be able to evaluate whether proposals for the enzyme-catalyzed mechanisms are reasonable. For the pH-independent hydrolysis of phosphate diesters, several kinetically equivalent mechanisms are plausible, including hydroxide attack on the neutral phosphate. However, it is very difficult to measure the rate of this reaction directly by experimental methods, so it has been evaluated by examining the rate of hydrolysis of neutral phosphate triesters, where a methyl group has replaced a proton. This may not be an accurate model of the neutral phosphate diester and does not provide information about a reaction pathway that is concerted with nucleophilic attack to generate a similar phosphorane. We have carefully mapped out free energy surfaces for both hydroxide and water attack on the dineopentyl phosphate anion and for water attack on the neutral diester. In doing so, we have accurately reproduced existing experimental data and demonstrate that water attack proceeds through an associative mechanism with proton transfer to the phosphate to generate a phosphorane intermediate. Our data show that the substrate-as-base mechanism is viable for phosphate ester hydrolysis, which may have important implications for the studies of phosphate ester hydrolysis by enzymes.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Computer Simulation
  • Hydrogen-Ion Concentration
  • Hydrolysis
  • Hydroxides / chemistry
  • Kinetics
  • Models, Chemical
  • Molecular Structure
  • Organophosphorus Compounds / chemistry*
  • Protons
  • Water / chemistry*

Substances

  • Hydroxides
  • Organophosphorus Compounds
  • Protons
  • dineopentyl phosphate
  • Water