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Project Title: The Allosteric Regulation of Caenorhabditis elegans Acyl-CoA Oxidase by ATP

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     Project Description: The nematode C. elegans secretes ascarosides, a class of pheromones worms use to communicate with each other. The ascarosides consist of an ascarylose core structure and a fatty acid-derived side chain. The side chains of the ascarosides are shortened by peroxisomal β-oxidation cycles. Acyl-CoA oxidase is involved in the rate limiting step of β-oxidation and installs a double bond at the α-β position of the ascaroside CoA. Previously, the crystal structure of C. elegans Acyl-CoA Oxidase 1.1 (ACOX-1.1) revealed the purified enzyme was bound not only to its FAD cofactor but also to ATP. Prior kinetic studies show that ACOX incubated without ATP or FAD gradually lost its activity. This loss could be blocked by the addition of ATP and FAD, but not FAD alone. We hypothesize that ATP functions as an allosteric regulator and that the binding of the enzyme to ATP alters the enzyme conformation and affects the affinity between the enzyme and its FAD cofactor. We will study the immediate effect of ATP and/or FAD addition on the activityof ACOX-1.1 using an HRP-coupled kinetic assay. To study the key residues involved in the allosteric network, we will utilize mutagenesis and assay the kinetics of the mutant enzyme.

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Presentations: UF Spring 2023 Undergraduate Research Symposiums