Alternative digestion approaches improve histone modification mapping by mass spectrometry in clinical samples

ABSTRACT: Profiling histone post-translational modifications (PTMs) in clinical samples holds great potential for the identification of epigenetic biomarkers and the discovery of novel epigenetic targets. We have recently developed a battery of mass spectrometry (MS)-based approaches to analyze histone PTMs in different types of primary samples. However, most of these protocols rely on SDS-PAGE separation following histone enrichment in order to eliminate detergents and isolate histones from other proteins present in the sample. As a consequence, many proteolytic enzymes, whose performance is poor in gel, cannot be used, limiting the digestions options for clinical samples, and hence the modification coverage. In this study, we used a simple procedure involving acetone protein precipitation followed by histone enrichment through a C18 StageTip column to obtain histone preparations suitable for various in solution digestion protocols.

INSTRUMENT(S): Q Exactive HF

ORGANISM(S): Homo Sapiens (human)

TISSUE(S): Leukemia Cell Line, Ovarian Serous Carcinoma Cell, Breast Cancer Cell

DISEASE(S): Acute Leukemia,Malignant Neoplasm Of Ovary,Breast Cancer

SUBMITTER: Roberta Noberini  

LAB HEAD: Tiziana Bonaldi

PROVIDER: PXD009946 | Pride | 2010-10-08

REPOSITORIES: Pride