July 01, 2016 at 12:18 PM

Prof Mark Dickman and colleagues have recently published work developing a  two dimensional-liquid chromatography (2D-LC) based approach for the identification and quantification of histone post translational modifications (PTMs) in conjunction with mass spectrometry analysis. Histone  PTMs  play key roles in regulating eukaryotic gene expression.  Epigenetic modifications are increasingly recognized as playing an important role in the pathogenesis of infectious diseases. Therefore, analysing changes in histone PTMs is important to study the role of epigenetics in the pathogenesis of infectious diseases. 

Mass spectrometry (MS) has emerged as a powerful method to identify and characterise histone PTMs. MS allows the unbiased identification and quantification of multiple global histone PTMs including combinations in a single experiment. In this study  a porous graphitic carbon stationary phase in the first dimension and a C18 stationary phase in the second dimension interfaced with mass spectrometry was used to analyse global levels of histone post translational modifications in human primary monocyte-derived macrophages. The results demonstrated that 84 different histone peptide proteoforms, with modifications at 18 different sites including combinatorial marks were identified, representing an increase in the identification of histone peptides by 65% and 51% compared to two different 1D-LC approaches on the same mass spectrometer.

This work was performed in collaboration with Prof Robert Read and Prof David Dockrell at the Department of Infection, Immunity & Cardiovascular Disease the University of Sheffield Medical School.

Read the published article here.