title: Comparative theoretical studies of the phosphomonoester hydrolysis mechanism by purple acid phosphatases.
abstract: We present here the first ONIOM (our own n-layered integrated molecular orbital + molecular mechanics method) studies of a purple acid phosphatase enzyme. Our study focused on the structures of the red kidney bean PAP (kbPAP) complexed with phosphate and with phenyl phosphate and on the mechanism of the phenyl phosphate hydrolysis by the enzyme. Density functional theory (DFT) calculations were also performed using models of different sizes for comparison purpose. Results show that the inclusion of three histidine residues, His202, His295, and His296, with their protein surrounding, is crucial to properly describe the coordination of the substrates. They induce a conformation with the substrate closer to the nucleophilic mu-hydroxyde bridge. In the mechanistic study, a transition state is stabilized by a strong hydrogen bond between His202 and the leaving group of the substrate. Consequently, a smaller value for the activation energy barrier is obtained from DFT calculations including this histidine to the same calculations without this histidine. Using the ONIOM method, this activation energy barrier is even more reduced. So the mechanism, which considers the hydroxo group bridging the two metal ions as nucleophile, becomes really convincing, contrary to the results obtained with a small model at the DFT level.
authors.affiliation: DCM, Equipe Chimie Théorique, UMR CNRS 5250, ICMG, FR CNRS, Université J. Fourier, BP. 53, 38041 Grenoble Cedex, France.
authors.forename: M
authors.initials: M
authors.lastname: Retegan
authors.forename: A
authors.initials: A
authors.lastname: Milet
authors.forename: H
authors.initials: H
authors.lastname: Jamet
mesh_terms: D000135:Acid Phosphatase / Q000737:chemistry* / Q000378:metabolism*; D018360:Crystallography, X-Ray; D006023:Glycoproteins / Q000737:chemistry* / Q000378:metabolism*; D006868:Hydrolysis; D008958:Models, Molecular; D009943:Organophosphorus Compounds / Q000737:chemistry / Q000378:metabolism*; D027805:Phaseolus / Q000201:enzymology; D010710:Phosphates / Q000737:chemistry / Q000378:metabolism; D011487:Protein Conformation; D011789:Quantum Theory
chemical_list: D006023:Glycoproteins; D009943:Organophosphorus Compounds; D010710:Phosphates; C091137:purple acid phosphatase; D000135:Acid Phosphatase