V5-TurboID-tagged Drosophila BubR1 and its cleavage variants were transiently expressed in Drosophila S2 cells in two biological replicates. Cells were treated with 500 µM biotin for 30 min for protein biotinylation. Biotinylated protein-containing lysate was subjected to FG-NeutrAvidin beads purification followed by on-beads trypsin digestion. Digested peptides were desalinated and purified using GL-tip SDB. Purified peptides were subjected to LC-MS/MS analysis.