Analysis of histone modifications from tryptic peptides of deuteroacetylated isoforms

Elisabeth Hersman, Dwella M. Nelson, Wendell P. Griffith, Christine Jelinek, Robert J. Cotter

Research output: Contribution to journalArticle

Abstract

The in vitro deuteroacetylation of histones obtained from biological sources has been used previously in bottom-up mass spectrometry analyses to quantitate the percent of endogenous acetylation of specific lysine sites and/or peptides. In this report, derivatization of unmodified lysine residues on histones is used in combination with high performance mass spectrometry, including combined HPLC MS/MS, to distinguish and quantitate endogenously acetylated isoforms occurring within the same tryptic peptide sequence and to extend this derivatization strategy to other post-translational modifications, specifically methylation, dimethylation and trimethylation. The in vitro deuteroacetylation of monomethylated lysine residues is observed, though dimethylated or trimethylated residues are not derivatised. Comparison of the relative intensities ascribed to the deuteroacetylated and monomethylated species with the deuteroacetylated but unmethylated analog, provides an opportunity to estimate the percent of methylation at that site. In addition to the observed fragmentation patterns, the very high mass accuracy available on the Orbitrap mass spectrometer can be used to confirm the structural isoforms, and in particular to distinguish between trimethylated and acetylated species.

Original languageEnglish (US)
Pages (from-to)5-16
Number of pages12
JournalInternational Journal of Mass Spectrometry
Volume312
DOIs
StatePublished - Feb 15 2012

Keywords

  • Acetylation
  • Histones
  • Methylation
  • Orbitrap
  • Proteomics

ASJC Scopus subject areas

  • Instrumentation
  • Condensed Matter Physics
  • Spectroscopy
  • Physical and Theoretical Chemistry

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