Mutagenesis and Purification of Bacterial Carboxylesterase from Francisella tularensis

Indianapolis, IN

Palmitoylation is a lipid modification that is added to proteins and controls the localization of proteins in cells. Cycling of proteins through different palmitoylation states is a central method for controlling protein activity in a cell. Misregulation of palmitoylation is also linked to the development of cancer. I have developed methods for measuring palmitoylation in two different bacteria, the model bacterium E. coli and a metabolically diverse bacterium P. putida. Using a mimic of palmitic acid, all palmitoylated proteins in the two bacteria can be tagged and then visualized by chemical attachment of the palmitoylated proteins to a colored tag. This method is called click chemistry. Click chemistry involves the reaction between an azide and an alkyne that is a biologically inactive reaction. The results of the research will provide tools for studying palmitoylation in bacteria and will identify targets for future antibacterial and cancer treatments.