Role of the conserved threonine 309 in mechanism of oxidation by cytochrome P450 2D6

P.H.J. Keizers, L.H. Schraven, C. de Graaf, M. Hidestrand, M. Ingelman-Sundberg, B.M. van Dijk, N.P.E. Vermeulen, J.N.M. Commandeur

Research output: Contribution to JournalArticleAcademicpeer-review

Abstract

Based on sequence alignments and homology modeling, threonine 309 in cytochrome P450 2D6 (CYP2D6) is proposed to be the conserved I-helix threonine, which is supposed to be involved in dioxygen activation by CYPs. The T309V mutant of CYP2D6 displayed a strong shift from O-dealkylation to N-dealkylation reactions in oxidation of dextromethorphan and 3,4- methylenedioxymethylamphetamine. This may be explained by an elevated ratio of hydroperoxo-iron to oxenoid-iron of the oxygenating species. In consistence, using cumene hydroperoxide, which directly forms the oxenoid-iron, the T309V mutant again selectively catalyzed the O-dealkylation reactions. The changed ratio of oxygenating species can also explain the decreased activity and changed regioselectivity that were observed in 7-methoxy-4-(aminomethyl)-coumarin and bufuralol oxidation, respectively, by the T309V mutant. Interestingly, the T309V mutant always showed a significantly increased, up to 75-fold, higher activity compared to that of the wild-type when using cumene hydroperoxide. These results indicate that T309 in CYP2D6 is involved in maintaining the balance of multiple oxygenating species and thus influences substrate and regioselectivity. © 2005 Elsevier Inc. All rights reserved.
Original languageEnglish
Pages (from-to)1065-74
JournalBiochemical and Biophysical Research Communications
Volume338
Issue number2
DOIs
Publication statusPublished - 2005

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