Characterization of an acyl-CoA dehydrogenase of unknown function

Details

Supervisors

Bommer, Guido ; Morsomme, Pierre

Faculty

Faculté des bioingénieurs

Degree label

Master [120] : bioingénieur en chimie et bioindustries, à finalité spécialisée

Abstract

Fatty acids play a central role in human metabolism, in cellular signaling and as structural component in membrane lipids. While fatty acid metabolism has been extensively studied, there are still some human enzymes of unknown function that resemble enzymes involved in lipid metabolism. In the present study, I tried to glean insights into the function of a putative human acyl-CoA dehydrogenase (ACAD), whose function is still unknown. The goal of my master’s thesis was to test whether related bacterial proteins might take over the function of this ACAD. Using a combination of in vitro and cell culture experiments, where metabolites were analyzed by LC-MS, we proceeded in three steps. First, we purified and characterized the enzymatic activity of purified bacterial proteins. Second, we tested whether we could drive expression of these proteins in mitochondria. Third, we tested whether expression of bacterial proteins could rescue metabolic changes observed in human knockout cell lines. In these experiments we found evidence for a functional overlap between the bacterial and the mammalian enzymes. This will form the basis for future investigations that will hopefully reveal the physiological function of the ACAD under investigation.