Lipoxygenases (LOs) are enzymes that catalyze the incorporation of dioxygen into 1,4-cis,cis-pentadiene containing fatty acids, and are widely distributed throughout the plant and animal kingdoms. The activity of the enzyme has been implicated in plant germination, human asthma and arthritis. The active site of the enzyme contains an essential iron atom. Two distinct mechanisms for LO catalysis have been proposed. The first involves a radical based mechanism in which the active site ferric ion oxidizes the 1,4-diene of linoleic acid (LA) to a fatty acid radical. The second mechanism uses the ferric iron to facilitate de-protonation of the substrate by coordinating the resulting carbonation to form an organometallic intermediate. Thus, the key difference in the two mechanisms is the role of the iron center and how it activates the proton abstraction. This proposal involves spectroscopic and kinetic studies aimed at investigating the structure and function of the active site iron in both soybean and human lipoxygenases. In addition, a number of active site mutants designed to affect the electronic structure of the iron and its reactivity will be studied. Spectroscopic techniques (e.g., EPR, MCD, EXAFS) will be used to determine the changes in the iron coordination and/or electronic environment. These studies will be correlated to kinetic investigations that address the effect of the structural changes on reactivity as well as the effects of pH, viscosity and isotopic substitutions. The information obtained will be used to determine a unified theory of the structure and function of the Fe center in catalyzing the oxidation reactions.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
First Independent Research Support & Transition (FIRST) Awards (R29)
Project #
5R29GM056062-03
Application #
2910348
Study Section
Metallobiochemistry Study Section (BMT)
Project Start
1997-05-01
Project End
2002-04-30
Budget Start
1999-05-01
Budget End
2000-04-30
Support Year
3
Fiscal Year
1999
Total Cost
Indirect Cost
Name
University of California Santa Cruz
Department
Chemistry
Type
Schools of Arts and Sciences
DUNS #
City
Santa Cruz
State
CA
Country
United States
Zip Code
95064
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Hoobler, Eric K; Rai, Ganesha; Warrilow, Andrew G S et al. (2013) Discovery of a novel dual fungal CYP51/human 5-lipoxygenase inhibitor: implications for anti-fungal therapy. PLoS One 8:e65928
Hoobler, Eric K; Holz, Charles; Holman, Theodore R (2013) Pseudoperoxidase investigations of hydroperoxides and inhibitors with human lipoxygenases. Bioorg Med Chem 21:3894-9
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