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Heavy Methyl-SILAC Labeling Coupled with Liquid Chromatography and High-Resolution Mass Spectrometry to Study the Dynamics of Site-Specific Histone Methylation

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Gene Regulation

Part of the book series: Methods in Molecular Biology ((MIMB,volume 977))

Abstract

Histone lysine and arginine methylation involved in gene activation and silencing is dynamically regulated. However, partly limited to the research technologies previously available, the dynamics of global histone methylation on a site-specific basis have not been fully pursued. Heavy methyl-SILAC (Stable Isotope Labeling of Amino Acids in Cell Culture) labeling provides a remarkable signpost to distinguish the preexisting and newly generated methyl marks on histones. Using this technology coupled with quantitative LC-MS analysis make it possible to monitor changes in the dynamics of histone site-specific methylation. In this chapter, we comprehensively describe the experimental strategy to determine the dynamics of multiple histone methylated residues including SILAC labeling, histone extraction/purification and mass spectrometry analysis.

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Acknowledgments

B.A.G. gratefully acknowledges funding from a National Science Foundation grant (CBET-0941143), an Agilent Thought Leader award, an NSF Early Faculty CAREER award, and an NIH Innovator award (DP2OD007447) from the Office Of The Director, NIH.

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Authors and Affiliations

  1. Department of Molecular Biology, Princeton University, Princeton, NJ, USA

    Xing-Jun Cao, Barry M. Zee & Benjamin A. Garcia

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  1. Xing-Jun Cao
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  2. Barry M. Zee
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  3. Benjamin A. Garcia
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Editors and Affiliations

  1. , Department of Chemistry, Purdue University, Oval Drive 560, West Lafayette, 47907-2084, Indiana, USA

    Minou Bina

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Cao, XJ., Zee, B.M., Garcia, B.A. (2013). Heavy Methyl-SILAC Labeling Coupled with Liquid Chromatography and High-Resolution Mass Spectrometry to Study the Dynamics of Site-Specific Histone Methylation. In: Bina, M. (eds) Gene Regulation. Methods in Molecular Biology, vol 977. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-284-1_24

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  • DOI: https://doi.org/10.1007/978-1-62703-284-1_24

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  • Publisher Name: Humana Press, Totowa, NJ

  • Print ISBN: 978-1-62703-283-4

  • Online ISBN: 978-1-62703-284-1

  • eBook Packages: Springer Protocols

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