Intrinsic isotope effects suggest that the reaction coordinate symmetry for the cytochrome P-450-catalyzed hydroxylation of octane is isozyme independent

Publisher Website
Google Scholar
Abstract
The mechanism of the .omega.-hydroxylation of octane by three catalytically distinct, purified forms of cytochrome P-450, namely, P-450b, P-450c, and P-450LM2, was investigated by using deuterium isotope effects. The deuterium isotope effects associated with the .omega.-hydroxylation of octane-1,1,1,-2H3, octane-1,8-2H2, and octane-1,1,8,8-2H4 by all three isozymes were determined. From these data the intrinsic isotope effects were calculated and separated into their primary and secondary components. The primary intrinsic isotope effect for the reaction ranged from 7.69 to 9.18 while the secondary intrinsic isotope effect ranged from 1.13 to 1.25. Neither the primary nor secondary isotope effect values were statistically different for any of the isozymes investigated. These data are consistent with a symmetrical transition state for a mechanism involving initial hydrogen atom abstraction followed by hydroxyl radical recombination which is essentially independent of the specific isozyme catalyzing the reaction. It is concluded that (1) in general the porphyrin-[FeO]3+ complex behaves as a source of a triplet-like oxygen atom, (2) the regioselectivity for the site of oxidation is dictated by the apoprotein of the specific isozyme of cytochrome P-450 catalyzing the reaction, and (3) the maximum primary intrinsic isotope effect for any cytochrome P-450 catalyzed oxidation of a carbon center is about 9, assuming no tunneling effects.