Immunoprecipitation of mitochondrial ribosome complex of Toxoplasma gondii

Mitochondrial ribosomes in Apicomplexan parasite Toxoplasma gondii are expected to be divergent from all the other known mitoribosomes owing to the extremely fragmented and reduced rRNAs. To elucidate the structure and composition of these mitoribosomes, we performed immunoprecipitation of the FLAG epitope tagged bL12m subunit of mitoribosome using anti-FLAG M2 agarose beads. The elute was used to freeze CryoEM grids for single particle analysis and bound beads were subjected to Liquid chromatography–mass spectrometry (LC–MS) to analyse the co-immunoprecipitated proteins.