Emerging mass spectrometry-based technologies for analyses of Chromatin changes: Analysis of histones and histone modifications

Brinda Shah, Rachel L. Kozlowski, Jun Han, Christoph H. Borchers

Research output: Chapter in Book/Report/Conference proceedingChapterpeer-review

5 Citations (Scopus)

Abstract

Mass spectrometry (MS) is rapidly becoming an indispensable tool for the analysis of posttranslational modifications (PTMs) of proteins, and particularly histone PTMs that regulate physiological processes. The more traditional bottom-up approach of searching for modifications on peptides rather than intact proteins (top-down) has proven useful for finding phosphorylation, acetylation, and ubiquitination sites. With the use of modern instrumentation and various MS-based techniques, peptides and their PTMs can be characterized in a high-throughput manner while still maintaining high sensitivity and specificity. In complement to bottom-up MS, recent advances in MS technology, such as high-field Fourier transform ion cyclotron resonance (FTICR)-mass spectrometry, have permitted the study of intact proteins and their modifications. On-line and off-line protein separation instruments coupled to FTICR-MS allow the characterization of PTMs previously undetectable with bottom-up approaches. The use of unique fragmentation techniques in FTICR-MS provides a viable option for the study of labile modifications. In this chapter, we provide a detailed description of the analytical tools - mass spectrometry in particular - that are used to characterize modifications on peptides and proteins. We also examine the applicability of these mass spectrometric techniques to the study of PTMs on histones via both the bottom-up and top-down proteomics approaches.

Original languageEnglish
Title of host publicationSeed Dormancy
Subtitle of host publicationMethods and Protocols
EditorsAllison R. Kermode
Pages259-303
Number of pages45
DOIs
Publication statusPublished - 2011
Externally publishedYes

Publication series

NameMethods in Molecular Biology
Volume773
ISSN (Print)1064-3745

Keywords

  • Acetylation
  • Chromatin
  • Epigenetics
  • FTICR-MS
  • Histones
  • Mass spectrometry
  • Posttranslational modification
  • Top-down proteomics

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