Lysyl oxidase is a copper-dependent amine oxidase that plays an important role in catalyzing the oxidation of amino groups in the proteins, collagen and elastin. The catalyzed process results in aldehyde molecules, which is essential in the production of the crosslinkages in these proteins. There are few studies involving the use of native lysyl oxidase from bovina aorta, which provides structural properties, specifically the connectivity of the five-disulfide linkages. The main focus of this study is to investigate the role each disulfide bond has on lysyl oxidase. A LOX mutant, C238S, was generated via PCR mutagenesis program. Overexpression of this mutant yielded 16.01mg of enzyme per liter of media. Total copper incorporation is currently being calculated. There was no catalytic activity for this mutant when compared to the wildtype. A redox cycling assay was performed, which confirms that a quinone is present even when one disulfide linkage was removed.

Evaluation of the role of disulfide crosslinkages in lysyl oxidase via site-directed mutagenesis

245th National Meeting (2013)

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