In mammals, prostaglandin H2 synthase (PGHS) catalyzes the critical function of synthesizing prostaglandins, which have an important role in signaling the body’s inflammatory response. A bacterial homolog predicted to have PGHS activity was discovered in the beta-proteobacterium Nitrosomonas europeae. This enzyme has been shown to have the lipid peroxidase activity associated with PGHS, but not its cyclooxygenase activity. In this study, site-directed mutagenesis was performed on the Nitrosomonas lipid peroxidase. Six amino acids in the active site of the Nitrosomonas peroxidase, predicted to be critical in its enzymatic activity, substrate binding, and cofactor binding, were systematically altered for functional comparison. Mutant proteins were overexpressed in E. coli BL21 cells and purified using an affinity resin. The purified Nitrosomonas mutants were characterized by kinetic analysis to examine the role of the modified amino acids on the Nitrosomonas PGHS enzymatic activity. Results obtained from this study will guide future explorations into structure-function relationships.

Site-directed mutagenesis of a lipid peroxidase from Nitrosomonas europeae

Macromolecular Chemistry: The Second Century

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ACS Spring 2020 National Meeting & Expo

The official archive and repository for all ACS Meetings & Expositions abstracts and select posters or presentations.