3P-01 PDF
Novel strategy for lysine acetylation discovery in acetylomics
Among 20 amino acids constituting protein, lysine is a hot spot of PTM such as methylation, ubiquitination and acetylation. Many PTMs occur on this residue and control the protein's function in a sophisticated manner. Above all, acetylation is the most ubiquitous and is known to regulate various cellular functions including metabolism, mitosis and aging. In order to understand the entire acetylation events as one single system, it is required to identify not only lysine acetylated proteins but also substrate-enzyme (lysine acetyl transferase (KAT) and lysine deacetylase (KDAC)) relationships in a proteome-scale. However, the current strategy for acetylomics based on immunoprecipitation has disadvantages such as the bias of antibody to substrates and the contamination of non-acetylated peptides.
In this study, we developed a new approach to enrich lysine acetylated peptides without using antibody, in which digested peptides with chemically protected amino groups are deacetylated by KDAC, followed by biotinylation to enrich the target peptides. In order to validate this method, we applied it to a standard protein digest containing KDAC substrate peptides. As a result, the target peptides were enriched exclusively from the KDAC-treated sample, whereas we didn’t identify the peptides from the control sample without KDAC treatment.