Methods in Molecular Biology Volume 977, 2013, pp 299-313
Date: 24 Jan 2013

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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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.