Discussion

We have confirmed the expression of the secondary alternative oxidase (AOX2) in all three human-infective trypanosomatids.  Our protein, unlike the trypanosome alternative oxidase TAO, has conserved orthologues in T. cruzi and Leishmania spp., and importantly is not present in mammalian cells.  We have shown that AOX2 is essential in all three of these parasites, making it an attractive multi-trypanosomatid drug target.  We have confirmed the protein to be localized to the mitochondrion and are investigating the effects of over- and under-expression on mitochondrial respiration.  We have successfully solubilized and purified recombinant AOX2 for determination of enzymatic activity and identification of potential inhibitors using our in-house focused natural product-like library and a fragment-based screen.