4 Year Wellcome Trust PhD Programme: Probing the Golgi apparatus proteome of the human parasite Trypanosoma brucei

The Golgi apparatus is the glycoprotein processing and targeting centre for eukaryotic cells. This multi-compartment organelle contains the glycohydrolases and glycosyltransferases that remodel and extend glycoprotein and glycolipid glycans prior to their sorting and delivery to the plasma membrane and lysosomes.
The parasitic protozoan organism Trypanosoma brucei causes human African trypanosomiasis (African sleeping sickness) in sub-Saharan Africa and is related to other kinetoplastid parasites of human and veterinary importance, such as T. vivax, T. congolense, T. cruzi and the leishmania. These trypanosomatid parasites are all highly dependent on their cell surface glycoproteins and glycolipids for survival and infectivity.
While bioinformatics and functional genomics has given us a partial ‘parts list’ and some precise information on the glycosylation machinery of T. brucei, we are still missing substantial amounts of information about the composition and molecular mechanisms taking place in the Golgi apparatus. This project will take advantage of developments in the Ferguson lab in stable isotope in cell culture (SILAC) quantitative proteomics and its application to defining high-confidence organelle proteomes to define a high-confidence T. brucei Golgi apparatus proteome. The resulting data will be mined by bioinformatics and potential therapeutically amenable pinch-points in glycoprotein processing and/or transport will be validated by genetic manipulation. Such targets may enter drug discovery via our Drug Discovery Unit (http://www.drugdiscovery.dundee.ac.uk/ ).
The project will provide a thorough training in parasite cultivation and genetic manipulation, quantitative proteomics and mass spectrometry-based glycomics.