Sensitive, Specific and Quantitative FT-ICR Mass Spectrometry of Combinatorial Post-Translational Modifications in Intact Histone H4

Logan Mackay, Bernard Ramsahoye, Karl Burgess, Ken Cook, Stefan Weidt, James Creanor, David Harrison, Pat Langridge-Smith, Ted Hupp, Larry Hayward

Research output: Contribution to journalArticlepeer-review

Abstract

We describe a quantitative Fourier transform ion cyclotron resonance mass spectrometric (FTICR MS) analysis of the relative proportions of post-translational modification states (PTMs) of core histones in cultured cells and tissues. A novel preseparation process using a monolithic column interfaced to a 12 T FTICR MS equipped with electron capture dissociation (ECD) yields very high mass accuracy spectra, allowing direct assignment of the PTMs present in the dominant modification states of intact H4, resolving a well recognized ambiguity between trimethylation and acetylation states. By eliminating preseparation, we also obtain a highly quantitative analysis of the distribution of H4 PTMs. Rapid, extensive, and reversible effects on PTMs induced by a histone deacetylase inhibitor indicate that H4 and other core histones are accessible to modification throughout the chromatin, not just in regions of active transcription. These methods provide tools for analysis of the histone code and its role in chromatin function.
Original languageEnglish
Pages (from-to)4147-4153
JournalAnalytical Chemistry
Volume80
Issue number11
DOIs
Publication statusPublished - Jun 2008

Fingerprint

Dive into the research topics of 'Sensitive, Specific and Quantitative FT-ICR Mass Spectrometry of Combinatorial Post-Translational Modifications in Intact Histone H4'. Together they form a unique fingerprint.

Cite this