Proteomic analyses of a bi-lobed structure in Trypanosoma brucei

The position and function of the Golgi apparatus are tightly coupled with the microtubule-organizing centers (MTOCs) that play important roles in cell growth and polarity. In the unicellular parasite Trypanosoma brucei, the single Golgi apparatus is adjacent to a novel, bilobed structure located at the proximal base of the flagellum, near to the basal body that nucleates the flagellar axoneme. The duplication and segregation of the bilobed structure are tightly coupled to the duplication and segregation of Golgi, ER exit site, basal bodies, and flagellum, suggesting a role of this unique structure in the precise positioning, biogenesis, and inheritance of these single-copied structures during the cell cycle of procyclic T. brucei. Here, we describe in details two different isolation and proteomic methods to characterize the protein components present on or associated with the bilobed structure, which allow further understanding of its function and association with other membranous and cytoskeletal organelles in the parasite.