C−H Bond Activation by a Ferric Methoxide Complex:  Modeling the Rate-Determining Step in the Mechanism of Lipoxygenase

Abstract

Lipoxygenases are mononuclear non-heme iron enzymes that regio- and stereospecifcally convert 1,4-pentadiene subunit-containing fatty acids into alkyl peroxides. The rate-determining step is generally accepted to be hydrogen atom abstraction from the pentadiene subunit of the substrate by an active ferric hydroxide species to give a ferrous water species and an organic radical. Reported here are the synthesis and characterization of a ferric model complex, [Fe(III)(PY5)(OMe)](OTf)(2), that reacts with organic substrates in a manner similar to the proposed enzymatic mechanism. The ligand PY5 (2,6-bis(bis(2-pyridyl)methoxymethane)pyridine) was developed to simulate the histidine-dominated coordination sphere of mammalian lipoxygenases. The overall monoanionic coordination provided by the endogenous ligands of lipoxygenase confers a strong Lewis acidic character to the active ferric site with an accordingly positive reduction potential. Incorporation of ferrous iron into PY5 and subsequent oxidation yields a stable ferric methoxide species that structurally and chemically resembles the proposed enzymatic ferric hydroxide species. Reactivity with a number of hydrocarbons possessing weak C-H bonds, including a derivative of the enzymatic substrate linoleic acid, scales best with the substrates' bond dissociation energies, rather than pK(a)'s, suggesting a hydrogen atom abstraction mechanism. Thermodynamic analysis of [Fe(III)(PY5)(OMe)](OTf)(2) and the ferrous end-product [Fe(II)(PY5)(MeOH)](OTf)(2) estimates the strength of the O-H bond in the metal bound methanol in the latter to be 83.5 +/- 2.0 kcal mol(-1). The attenuation of this bond relative to free methanol is largely due to the high reduction potential of the ferric site, suggesting that the analogously high reduction potential of the ferric site in LO is what allows the enzyme to perform its unique oxidation chemistry. Comparison of [Fe(III)(PY5)(OMe)](OTf)(2) to other coordination complexes capable of hydrogen atom abstraction shows that, although a strong correlation exists between the thermodynamic driving force of reaction and the rate of reaction, other factors appear to further modulate the reactivity.

A synthetic precedent for the [Fe IV 2 (μ-O) 2 ] diamond core proposed for methane monooxygenase intermediate Q

“…8-10). In all three cases, DHA was oxidized and kinetic isotope effects (KIEs) Ͼ 9 were observed with DHA-d 4 as substrate ( Table 2), indicating that hydrogen atom abstraction is the rate-determining step for the oxidations (32,33). Analysis of the oxidation products showed that anthracene was produced in Ϸ50% yield for 1b and 4b, as deduced from its characteristic UV bands (SI Figs.…”

The effect of traumatic acid on the growth, metabolite content and antioxidant activity in Wolffia arrhiza (L.) Wimm. (Lemnaceae)

“…The last stage of the TA biosynthesis pathway is the non-enzymatic oxidation of traumatin into traumatic acid. TA synthesis is induced and regulated mainly by environmental stress factors, especially wounding, but also by high and low temperatures, UV, or osmotic shock (Zimmerman and Coudron 1979, Siedow 1991, Stelt et al 2000, Goldsmith et al 2002, Howe and Schilmiller 2002.…”

Differential quantum tunneling contributions in nitroalkane oxidase catalyzed and the uncatalyzed proton transfer reaction

“…Although tantalizing, the relatively small difference in the observed KIEs is not sufficient to conclude that there is a greater tunneling contribution in the enzymatic process than that of the uncatalyzed reaction in water. A number of experimental and theoretical studies suggested that the extent of tunneling was the same in several enzymes as in model reactions in solution (2,9,10), whereas other experiments showing differential tunneling behaviors may be attributed to different reaction mechanisms (2,(11)(12)(13). Nuclear tunneling cannot be measured directly, and the best experimental diagnostic of tunneling is through measurement of kinetic isotope effects (14).…”