TgCoq5 proximity proteome in mitochondria and cytosolic fractions using FbxO14 as spatial reference

Coenzyme Q or Ubiquinone (UQ) is an essential electron carrier and antioxidant. In yeast and mammals, several enzymes involved in the UQ synthesis form a protein complex in the mitochondrial matrix. Blast searches of the Toxoplasma gondii proteome failed to reveal several essential enzymes of the ubiquinone synthesis pathway. With the aim to identify those missing enzymes, we generated mutant strain expressing TurboID at the C-terminus of TgCoq5, one of the enzymes we have validated to be involved in UQ synthesis in T. gondii. Using a strategy combining biotinylation of TgCoq5-TurboID and FbxO14-TurboID, used as spatial reference, and subcellular fractionation. We investigated the proximal proteome of TgCoq5 and FbxO14 in the mitochondrial enriched fractions, P2, and cytosolic enriched fractions, S4.