Mapping Trypanosoma Protein Interactome by Proximity-Dependent Biotinylation

Proximity labeling techniques, such as BioID and TurboID, have emerged as modern tools for studying protein-protein interactions (PPIs) in cells. TurboID is a highly efficient biotin ligase that biotinylates proximal proteins in living cells when fused to a bait protein. These biotinylated proteins are enriched using a pull-down assay with streptavidin beads and identified through proteomic analysis, revealing the interactome of the target protein. Here, we describe proximity labeling using TurboID fused to Trypanosoma brucei PEX19, an essential cytosolic protein and a receptor for newly synthesized glycosomal membrane proteins, to identify its interactome. Understanding these interactions can be crucial for identifying new drug targets to combat these parasites. This powerful approach for mapping interactomes can be applied broadly to other organisms, including plants, yeast, and mammalian systems.