1P-14 PDF
Quantitative Evaluation for Residue-selectivity of Histone Acetylation by Histone Acetyl Transferases (HATs) and Artificial Catalysts Using LC–MS/MS
Histone acetylation is crucial for epigenetic mechanisms involved in diverse biological processes and diseases, which is physiologically catalyzed by histone acetyltransferases (HATs). Although studies on site selectivity of acetylation by HAT are necessary to understand epigenetic mechanisms, there are only a few reports on residue-selectivity investigated using MS/MS technique. On the other hand, we have developed artificial catalyst systems that could replace HATs, which also require the evaluation of residue-selectivity. Here we report on the semi-comprehensive quantitative analysis of lysine acetylation on recombinant nucleosome by using LC–MS/MS. Briefly, acetylated nucleosomes were derivatized with propionic anhydride, and the following digestion gave propionylated or acetylated peptides, whose peak areas are used for calculation of acetylation stoichiometry of each lysines. By using this method, we evaluated the residue specificity of HATs and our artificial catalyst systems.