title: Heavy methyl-SILAC labeling coupled with liquid chromatography and high-resolution mass spectrometry to study the dynamics of site-specific histone methylation.
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.
authors: Department of Molecular Biology, Princeton University, Princeton, NJ, USA.
authors: Xing-Jun
authors: XJ
authors: Cao
authors: Barry M
authors: BM
authors: Zee
authors: Benjamin A
authors: BA
authors: Garcia
mesh_terms: D000812:Anhydrides / Q000737:chemistry; D000818:Animals; D002247:Carbon Isotopes / Q000737:chemistry; D002458:Cell Fractionation; D002460:Cell Line; D002467:Cell Nucleus / Q000737:chemistry; D002851:Chromatography, High Pressure Liquid; D056148:Chromatography, Reverse-Phase; D006657:Histones / Q000737:chemistry* / Q000302:isolation & purification / Q000378:metabolism; D006801:Humans; D007553:Isotope Labeling / Q000379:methods*; D008715:Methionine / Q000737:chemistry; D008745:Methylation; D011422:Propionates / Q000737:chemistry; D011499:Protein Processing, Post-Translational; D059748:Proteolysis; D053719:Tandem Mass Spectrometry / Q000379:methods; D014357:Trypsin / Q000737:chemistry
chemical_list: D000812:Anhydrides; D002247:Carbon Isotopes; D006657:Histones; D011422:Propionates; D008715:Methionine; C096126:propionic anhydride; D014357:Trypsin