Human KAT2A/B-acetylome MudPIT

Comprehensive identification of acetylated proteins targeted by the lysine acetyltransferases KAT2A (GCN5) and KAT2B (PCAF), in human cells. KAT2A/B-dependent acetylated proteins were identified by comparing acetylated proteins in whole cell extract of control cells and cells in which KAT2A and KAT2B were simultaneously knocked down. The identification of acetylated proteins was performed using an enrichment-free approach. In order to do so, while facing the challenge of identifying low abundant acetylated peptides within a complex sample mixture, we have screened acetylated peptides starting from a large amount of material, equivalent to 50ug of protein digest (maximum loading capacity of our analytical chromatographic column). By analysing such a large amount of peptides, the level of acetylated peptides was then increased and thus acetylated peptides became easily detectable by mass spectrometry, without the need for enrichment.