Intracellular location of the early steps of the isoprenoid biosynthetic pathway in the trypanosomatids Leishmania major and Trypanosoma brucei

摘要: The isoprenoid biosynthetic pathway is a very complex route that entails multiple steps and generates high number of end-products are essential for cell viability such as sterols, dolichols, coenzyme Q, heme prenylated proteins. In parasites from the Trypanosomatidae family this provides new potential drug targets exploitation in search improved therapies, indeed compounds ketoconazole, aminobisphosphonates or terbinafine have been shown to antiprotozoal activity both vitro vivo. However, despite therapeutic importance pathway, subcellular compartmentalization different biosynthesis not known detail. Here we analysed intracellular location enzymes 3-hydroxy-3-methyl-glutaryl Coenzyme A (HMG-CoA) synthase (HMGS) mevalonate kinase (MVAK) Leishmania major promastigotes well Trypanosoma brucei procyclic bloodstream forms. For purpose generated specific polyclonal antibodies against highly purified recombinant proteins used those indirect immunofluorescence digitonin titration experiments. Results show sterol distributed compartments provide evidence indicating trypanosomatids production HMG-CoA acetyl generation occur mainly mitochondrion while further phosphorylation almost exclusively located glycosomes. Furthermore, determined peroxin 2 (PEX2) involved efficient targeting MVAK enzyme relocated cytosol upon depletion glycosomal matrix protein import.