I am a PhD student co-supervised by both Angus and Mike Ferguson, and funded by the Wellcome Trust. The aim of project is to further characterise the protozoan parasite, Trypanosoma brucei, utilising proteomic methodologies.
Trypanosoma brucei is a unicellular protozoan parasite that causes African sleeping sickness in humans and n’agana in livestock in sub-Saharan Africa. The trypanosomatid order also includes the parasites Trypanosoma cruzi (Chagas disease) and Leishmania major (Leishmaniasis). Sleeping sickness is estimated to cause ~10,000 deaths per year. Current treatments are expensive, difficult to administer and toxic.
Trypanosomatids are a major cause of global disease, however due to their large evolutionary divergence from intensively studied eukaryotic model organisms much of their unusual biology remains poorly characterised. Although genomic sequences have been produced for all three parasites, much is still unknown about protein function with 64% of identified genes annotated as “hypothetical” containing no known regions of homology with known proteins or domains.
To overcome the lack of functional annotation from genomic sequence, efforts are being made to provide experimental evidence for protein function, for example the development of genome wide gene knockdowns utilising RNA interference followed by phenotypic analysis or transcriptomic and proteomic analyses focusing on different aspects of parasite biology.
My project aims to build on this work by providing experimental evidence for function of proteins in T. brucei by determining putative interaction partners on a proteome wide scale. To achieve this analyses I will be separating complex trypanosome lysates by different chromatographic methods, followed by identification of proteins present in the eluted fractions collected. To identify putative interactions we then cluster the proteins identified based on the correlation between elution profiles determined by mass spectrometry.
Through identifying putative interactions of trypanosome proteins we may uncover function for “hypothetical” annotated proteins through “guilt by association” with other components with known function.
I also aim to compare different biological samples, different forms of the parasite or different cell cycle stages, for example, leading to an idea of how T. brucei modifies not only the composition, but the organisation of its proteome to enact different processes. The data produced will also act as a useful resource for other trypanosomatid researchers interested in finding potential interaction partners of proteins of interest, helping to uncover biological function.