title: 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.
authors.affiliation: Department of Chemistry, Stanford University, Stanford, California 94305, USA.
authors.forename: Christian R
authors.initials: CR
authors.lastname: Goldsmith
authors.forename: Robert T
authors.initials: RT
authors.lastname: Jonas
authors.forename: T Daniel P
authors.initials: TD
authors.lastname: Stack
mesh_terms: D018360:Crystallography, X-Ray; D003510:Cyclohexanes / Q000737:chemistry; D053138:Cyclohexenes; D005290:Ferric Compounds / Q000737:chemistry*; D006859:Hydrogen / Q000737:chemistry; D007700:Kinetics; D008084:Lipoxygenase / Q000737:chemistry* / Q000378:metabolism; D000432:Methanol / Q000031:analogs & derivatives* / Q000737:chemistry; D008958:Models, Molecular; D018716:Molecular Mimicry; D010084:Oxidation-Reduction; D010636:Phenols / Q000737:chemistry; D013816:Thermodynamics
chemical_list: D003510:Cyclohexanes; D053138:Cyclohexenes; D005290:Ferric Compounds; D010636:Phenols; C048401:1,4-cyclohexadiene; D006859:Hydrogen; C019584:2,4,6-tri-tert-butylphenol; D008084:Lipoxygenase; D000432:Methanol