Site-Link for identification of chromatinized proteome defined by site-specific histone acylations in living cells

By coupling the mass spectrometry, we developed the Site-Link strategy based on the GCE method for analyzing the interacting proteome of histone lysine acylations in living cells. As K* could mimic the natural lysine to be recognized by natural lysine synthase, histones bearing the site-specifically incorporated K*acyl, should also be also integrated into nucleosomes and further compacted into chromatin similar as Kacyl with cell passaging. The subsequent 365-nm light irradiation could activate the diazirine and covalently capture potential effector proteins on native chromatin in living cells. The effector proteins could be identified after gel-based proteomics and MS/MS analysis of the crosslinked histone-effector complexes.