101 Scopus citations

Abstract

Histone variants and posttranslational modifications (PTMs) are essential for epigenetic regulation of transcriptional expression. Single and/or combinatorial PTMs of histones play important roles in development and disease formation. Mass spectrometry (MS) has been a powerful tool to study histone variants and PTMs, as it not only can identify novel PTMs but also can provide quantitative measurement of a spectrum of histone variants and PTMs in the same sample. In this chapter, we employ a combination of chemical derivation and high-resolution MS to identify and quantify multiple histone variants and PTMs. Histones are acid extracted and modified with propionyl groups and subsequently produces suitable sizes of fragments for MS analysis by trypsin digestion. The newly generated N-termini of histone peptides can be differentially marked by stable isotope labeling in a second reaction of propionylation, which enables direct comparison between two different samples in the following MS analysis.

Original languageEnglish
Title of host publicationMethods in Enzymology
PublisherAcademic Press Inc.
Pages3-28
Number of pages26
DOIs
StatePublished - 2012

Publication series

NameMethods in Enzymology
Volume512
ISSN (Print)0076-6879
ISSN (Electronic)1557-7988

Keywords

  • Epigenetics
  • High-performance liquid chromatography
  • Histone posttranslational modifications
  • Mass spectrometry
  • Propionylation
  • Stable isotope labeling

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