MS-based proteomic characterization of Toxoplasma gondii GSK3 proxisome

In an effort to identify novel drug candidates against T. gondii parasites, an interesting compound was identified, allowing to inhibit tachyzoite growth without impacting the HFF monolayer. A forward genetic screen identifies TgGSK3 as the enzyme targeted by this molecule. To investigate the biological function of TgGSK3 and its regulatory mechanisms, we performed in vivo biotinylation using the proximity labeling approach employing BioID2 with TgGSK3 as the bait protein. To identify candidate interacting partners or substrates of TgGSK3, biotinylated proteins were enriched from lysates using streptavidin affinity purification. As a control, we conducted a mock experiment with parasites that do not express BioID2 fusion protein (RH ku80).