Identification of Khib substrates of histone regulated by NAT10

Having established NAT10’s regulatory role in lysine 2-hydroxyisobutyrylation, we aimed to identify the histone Khib sites targetedby NAT10. To this end, we extracted and digested core histones from cells with or without NAT10 depletion, followed by chemical derivatization of amine groups. The resulting peptides were analyzed by data-independent acquisition (DIA) mass spectrometry to detect the NAT10-modulated Khib sites.