Mass spectrometry analysis of kinetoplastid kinetochore proteins (KKT23 acetyltransferase and its interactor KKT22) and histone acetylation levels in Trypanosoma brucei

KKT23 is a kinetoplastid kinetochore protein that has structural similarity to the GCN5 acetyltransferase domain. Using a kinetoplastid parasite Trypanosoma brucei, here we performed immunoprecipitation and mass spectrometry of GFP-tagged KKT23 N-terminal fragment (2–150) to identify co-purifying proteins, and immunoprecipitation and mass spectrometry of H2B-GFP after KKT23 depletion to examine acetylation states of histones. We also performed in vitro acetylation assays on recombinant trypanosome nucleosomes using recombinant KKT23 protein, followed by mass spectrometry.