Authors

Discussion

Current treatments for kinetoplastid related diseases are inadequate and there is an urgent need for lead compounds that can be translated into safe, cheap, and easy to administer drugs. The causative agent of Human African Trypanosomiasis, Trypanosoma brucei, relies upon endocytosis and degradation of host macromolecules from the mammalian bloodstream to acquire metabolites vital for its proliferation and survival. The terminal compartment of the endocytic pathway, the lysosome is critical to this macromolecular digestion. However, very little is known about this organelle with only a handful of proteins characterised. To successfully study essential processes and identify new drug targets within it, the lysosome should be isolated. Standard cellular fractionation is ineffective unless a non-digestible macromolecule is employed which accumulates in T. brucei lysosomes and alters their density so they can be more easily purified. Sucrose gradient centrifugation produced a distinct fraction that contained electron dense particles comparable in morphology to lysosomes. Proteomic analysis of the lysosome-like particles identified known lysosomal markers and proteins with digestive, structural and transport related functions. We are utilising this approach to investigate lysosomal catabolic processes in detail and test compounds that will disrupt the function of this essential T. brucei organelle.