F. Guengerich, Grover P. Miller, I. H. Hanna
Sep 13, 2002
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0
Influential Citations
45
Citations
Quality indicators
Journal
The Journal of Biological Chemistry
Abstract
Cytochrome P450 (P450) 2D6 is involved in the oxidation of a large fraction (∼30%) of drugs used by humans and also catalyzes the O-demethylation of the model substrates 3- and 4-methoxyphenethylamine followed by subsequent ring hydroxylation to dopamine. Burst kinetics were not observed; rate-limiting step(s) must occur prior to product formation. Rates of reduction of ferric P450 2D6 were stimulated by 3- or 4-methoxyphenethylamine or the inhibitor quinidine; reduction is not the most rate-limiting step. The non-competitive intramolecular deuterium isotope effect, an estimate of the intrinsic isotope effect, for 4-methoxyphenethylamine O-demethylation was 9.6. Intermolecular non-competitive deuterium isotope effects of 3.1–3.8 were measured for k cat andk cat/K m for bothO-demethylation reactions, implicating at least partially rate-limiting C–H bond breaking. Simulation of steady-state kinetic data yielded a catalytic mechanism dominated by the rates of (i) Fe2+O 2 − protonation (plus O–O bond scission) and (ii) C–H bond breaking, consistent with the appearance of the spectral intermediates in the steady state, attributed to iron–oxygen complexes. However, all the rates of individual steps (or rates of combined steps) are considerably higher thank cat, and the contributions of several steps must be considered in understanding rates of the P450 2D6 reactions.