4-Hydroxyphenylpyruvate dioxygenase: a hybrid density functional study of the catalytic reaction mechanism.
Density functional calculations using the B3LYP functional has been used to study the reaction mechanism of 4-hydroxyphenylpyruvate dioxygenase. The first part of the catalytic reaction, dioxygen activation, is found to have the same mechanism as in alpha-ketoglutarate-dependent enzymes; the ternary enzyme-substrate-dioxygen complex is first decarboxylated to the iron(II)-peracid intermediate, followed by heterolytic cleavage of the O-O bond yielding an iron(IV)-oxo species. This highly reactive intermediate attacks the aromatic ring at the C1 position and forms a radical sigma complex, which can either form an arene oxide or undergo a C1-C2 side-chain migration. The arene oxide is found to have no catalytic relevance. The side-chain migration is a two-step process; the carbon-carbon bond cleavage first affords a biradical intermediate, followed by a decay of this species forming the new C-C bond. The ketone intermediate formed by a 1,2 shift of an acetic acid group rearomatizes either at the active site of the enzyme or in solution. The hypothetical oxidation of the aromatic ring at the C2 position was also studied to shed light on the 4-HPPD product specificity. In addition, the benzylic hydroxylation reaction, catalyzed by 4-hydroxymandelate synthase, was also studied. The results are in good agreement with the experimental findings.
Borowski
Tomasz
T
Department of Physics, Stockholm Center for Physics, Astronomy, and Biotechnology, Stockholm University, S-106 91, Stockholm, Sweden. borowski@physto.se
Bassan
Arianna
A
Siegbahn
Per E M
PE
D015111:4-Hydroxyphenylpyruvate Dioxygenase / Q000737:chemistry* / Q000378:metabolism*; D019342:Acetic Acid / Q000737:chemistry; D001665:Binding Sites; D002384:Catalysis; D018360:Crystallography, X-Ray; D004789:Enzyme Activation; D006900:Hydroxylation; D007501:Iron / Q000737:chemistry / Q000378:metabolism; D008958:Models, Molecular; D010100:Oxygen / Q000378:metabolism; D010667:Phenylpyruvic Acids / Q000378:metabolism; D017434:Protein Structure, Tertiary; D013379:Substrate Specificity
D010667:Phenylpyruvic Acids; C010590:4-hydroxyphenylpyruvic acid; D007501:Iron; D015111:4-Hydroxyphenylpyruvate Dioxygenase; D019342:Acetic Acid; D010100:Oxygen