title: The effects of substrate orientation on the mechanism of a phosphotriesterase.
abstract: While the underlying chemistry of enzyme-catalyzed reactions may be almost identical, the actual turnover rates of different substrates can vary significantly. This is seen in the turnover rates for the catalyzed hydrolysis of organophosphates by the bacterial phosphotriesterase OpdA. We investigate the variation in turnover rates by examining the hydrolysis of three classes of substrates: phosphotriesters, phosphothionates, and phosphorothiolates. Theoretical calculations were used to analyze the reactivity of these substrates and the energy barriers to their hydrolysis. This information was then compared to information derived from enzyme kinetics and crystallographic studies, providing new insights into the mechanism of this enzyme. We demonstrate that the enzyme catalyzes the hydrolysis of organophosphates through steric constraint of the reactants, and that the equilibrium between productively and unproductively bound substrates makes a significant contribution to the turnover rate of highly reactive substrates. These results highlight the importance of correct orientation of reactants within the active sites of enzymes to enable efficient catalysis.
authors.affiliation: Research School of Chemistry, Australian National University, Canberra, ACT 0200, Australia.
authors.forename: Colin J
authors.initials: CJ
authors.lastname: Jackson
authors.forename: Jian-Wei
authors.initials: JW
authors.lastname: Liu
authors.forename: Michelle L
authors.initials: ML
authors.lastname: Coote
authors.forename: David L
authors.initials: DL
authors.lastname: Ollis
mesh_terms: D001665:Binding Sites; D002384:Catalysis; D019295:Computational Biology; D006868:Hydrolysis; D008958:Models, Molecular; D015394:Molecular Structure; D044345:Phosphoric Triester Hydrolases / Q000737:chemistry* / Q000378:metabolism*; D017434:Protein Structure, Tertiary; D013379:Substrate Specificity
chemical_list: D044345:Phosphoric Triester Hydrolases