Characterization of neurohistone variants and post-translational modifications by electron capture dissociation mass spectrometry

Benjamin A. Garcia, Nertila Siuti, C. Eric Thomas, Craig A. Mizzen, Neil L. Kelleher

Research output: Contribution to journalArticle

Abstract

Post-translational modifications (PTMs) of histones are intimately involved in chromatin structure and thus have roles in cellular processes through their impact on gene activation or repression. At the forefront in histone PTM analysis are mass spectrometry-based techniques, which have capabilities to produce improved views of processes affected by chromatin remodeling via histone modifications. In this report, we take the first mass spectrometric look at histone variant expression and post-translational modifications from histones isolated from rat brain tissue. Analyses of whole rat brain identified specific histone H2A and H2B gene family members and several H4 and H3 post-translational modification sites by electron capture dissociation (ECD) mass spectrometry. We subsequently compared these results to selected rat brain regions. Major differences in the expression profiles of H2A and H2B gene family members or in the post-translational modifications on histone H4 were not observed from the different brain regions using a Top Down approach. However, "Middle Down" mass spectrometry facilitating improved characterization of the histone H3 tail (1-50 residues), revealed an enrichment of trimethylation on Lys9 from cerebellum tissue compared to H3 extracted from whole brain, cerebral cortex or hypothalamus tissue. We forward this study in honor of Professor Donald F. Hunt, whose pioneering efforts in protein and PTM analyses have spawned new eras and numerous careers, many exemplified in this special issue.

Original languageEnglish (US)
Pages (from-to)184-196
Number of pages13
JournalInternational Journal of Mass Spectrometry
Volume259
Issue number1-3
DOIs
StatePublished - Jan 1 2007

Keywords

  • Brain
  • Electron capture dissociation (ECD)
  • Histone
  • Mass spectrometry
  • Post-translational modification

ASJC Scopus subject areas

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

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