Comparative analysis of histone post-translational modification patterns obtained by mass spectrometry on intact proteins, Asp-N/Glu-C and tryptic peptides

摘要

The relative histone PTMs abundances measured on intact proteins, Asp-N/Glu-C and trypsin peptides were very similar whatever the method used. While bottom-up MS is perfectly suited to localize PTMs, middle-down and top-down MS enable 'long-range' combinatorial analysis of histone modifications. In certain instances, only the information obtained at the protein level enable variant discrimination. These approaches are complementary, the use of an integrated top-down/bottom-up approach is required for optimal characterization of histone PTMs. The developed UHPLC-MS methodology was demonstrated to be efficient and reproducible for assessing the global combinatorial modification profiles of core histones in a high-throughput manner whereas the two other approaches are more time-consuming.