Authors

Discussion

myo-Inositol is one of the nine naturally occurring inositol stereoisomers. It is ubiquitous amongst eukaryotes and acts as an essential metabolite with roles in signal transduction, membrane formation, and cellular physiology. In the protozoan parasite Trypanosoma cruzi—the causative agent of Chagas’ disease—myo-inositol acts as a precursor to phosphatidylinositol (PI), which is an essential component to membrane lipids. In addition, PI in turn is required for formation of inositol phosphorylceramide (IPC), various phosphoinositides, and glycophosphatidylinositol (GPI)-anchored mucin-type glycoproteins, which coats the parasite’s cell-surface allowing the parasite to participate in multiple essential steps in parasite-host interactions. In T. cruzi, myo-inositol is proposed to be both de novo synthesised as well as scavenged from the environment, however, the proteins involved in myo-inositol de novo synthesis have not been studied in T. cruzi. Therefore, the aim of this project is to genetically validate and biochemically characterise the putative inositol-3-phosphate synthase (TcINO1). This is done through recombinant expression and purification of TcINO1 for enzyme assays. Additionally, TcINO1 will be genetically manipulated in T. cruzi to determine essentiality and phenotype decreased myo-inositol de novo synthesis through lipid analysis.