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Seed Dormancy pp 259–303Cite as

Emerging Mass Spectrometry-Based Technologies for Analyses of Chromatin Changes: Analysis of Histones and Histone Modifications

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Part of the book series: Methods in Molecular Biology ((MIMB,volume 773))

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.

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Acknowledgements

This work was supported by a Genome Canada/Genome British Columbia Technology Development Grant and a platform grant from Genome Canada and Genome British Columbia.

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Authors and Affiliations

  1. Department of Biochemistry and Microbiology, and the University of Victoria – Genome British Columbia Protein Center, University of Victoria, Victoria, BC, Canada

    Brinda Shah, Rachel L. Kozlowski & Christoph H. Borchers

  2. University of Victoria - Genome British Columbia Protein Centre, University of Victoria, Victoria, BC, Canada

    Jun Han

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  1. Brinda Shah
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  2. Rachel L. Kozlowski
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  3. Jun Han
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  4. Christoph H. Borchers
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Correspondence to Christoph H. Borchers .

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  1. Dept. Biological Sciences, Simon Fraser Unviersity, University Drive 8888, Burnaby, V5A 1S6, British Columbia, Canada

    Allison R. Kermode

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Shah, B., Kozlowski, R.L., Han, J., Borchers, C.H. (2011). Emerging Mass Spectrometry-Based Technologies for Analyses of Chromatin Changes: Analysis of Histones and Histone Modifications. In: Kermode, A. (eds) Seed Dormancy. Methods in Molecular Biology, vol 773. Humana Press. https://doi.org/10.1007/978-1-61779-231-1_16

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  • DOI: https://doi.org/10.1007/978-1-61779-231-1_16

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