Analysis of core histones by liquid chromatography–mass spectrometry and peptide mapping

Histone acetylation and methylation are processes that are generally considered to play crucial roles in the chromatin-based regulatory mechanism. Characterization of the histones as well as their modification sites has become increasingly important. In this paper, the use of LC–MS and peptide mapping methods to analyze chicken core histones and identify the modification sites is reported. Microbore C4 HPLC separated the core histones into H2A, H2B, H3 and H4 using HFBA as the ion-pairing agent. The four subclasses of histones and their putative acetylated or methylated isoforms were identified by LC–MS simultaneously. MALDI-TOF and tandem mass spectrometry provided peptide mapping of the modification sites of the histones through trypsin digestion of the HPLC eluents. This approach is straightforward and prospective for further application in the field of chromatin research.