Oxygen activation by the noncoupled binuclear copper site in peptidylglycine alpha-hydroxylating monooxygenase. Reaction mechanism and role of the noncoupled nature of the active site.
Reaction thermodynamics and potential energy surfaces are calculated using density functional methods to investigate possible reactive Cu/O(2) species for H-atom abstraction in peptidylglycine alpha-hydroxylating monooxygenase (PHM), which has a noncoupled binuclear Cu active site. Two possible mononuclear Cu/O(2) species have been evaluated, the 2-electron reduced Cu(II)(M)-OOH intermediate and the 1-electron reduced side-on Cu(II)(M)-superoxo intermediate, which could form with comparable thermodynamics at the catalytic Cu(M) site. The substrate H-atom abstraction reaction by the Cu(II)(M)-OOH intermediate is found to be thermodynamically accessible due to the contribution of the methionine ligand, but with a high activation barrier ( approximately 37 kcal/mol, at a 3.0-A active site/substrate distance), arguing against the Cu(II)(M)-OOH species as the reactive Cu/O(2) intermediate in PHM. In contrast, H-atom abstraction from substrate by the side-on Cu(II)(M)-superoxo intermediate is a nearly isoenergetic process with a low reaction barrier at a comparable active site/substrate distance ( approximately 14 kcal/mol), suggesting that side-on Cu(II)(M)-superoxo is the reactive species in PHM. The differential reactivities of the Cu(II)(M)-OOH and Cu(II)(M)-superoxo species correlate to their different frontier molecular orbitals involved in the H-atom abstraction reaction. After the H-atom abstraction, a reasonable pathway for substrate hydroxylation involves a "water-assisted" direct OH transfer to the substrate radical, which generates a high-energy Cu(II)(M)-oxyl species. This provides the necessary driving force for intramolecular electron transfer from the Cu(H) site to complete the reaction in PHM. The differential reactivity pattern between the Cu(II)(M)-OOH and Cu(II)(M)-superoxo intermediates provides insight into the role of the noncoupled nature of PHM and dopamine beta-monooxygenase active sites, as compared to the coupled binuclear Cu active sites in hemocyanin, tyrosinase, and catechol oxidase, in O(2) activation.
Chen
Peng
P
Department of Chemistry, Stanford University, Stanford, California 94305, USA.
Solomon
Edward I
EI
D001665:Binding Sites; D019295:Computational Biology; D003300:Copper / Q000737:chemistry* / Q000378:metabolism; D006899:Mixed Function Oxygenases / Q000737:chemistry* / Q000378:metabolism; D008956:Models, Chemical; D008958:Models, Molecular; D009097:Multienzyme Complexes / Q000737:chemistry* / Q000378:metabolism; D010100:Oxygen / Q000737:chemistry* / Q000378:metabolism; D013816:Thermodynamics
D009097:Multienzyme Complexes; D003300:Copper; D006899:Mixed Function Oxygenases; C035189:peptidylglycine monooxygenase; D010100:Oxygen