Gene Regulation

Volume 977 of the series Methods in Molecular Biology pp 299-313


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

  • Xing-Jun CaoAffiliated withDepartment of Molecular Biology, Princeton University
  • , Barry M. ZeeAffiliated withDepartment of Molecular Biology, Princeton University
  • , Benjamin A. GarciaAffiliated withDepartment of Molecular Biology, Princeton University

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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.

Key words

Histone Methylation Dynamics Heavy methionine SILAC Quantitation Mass spectrometry