title: Separation and purification of multiply acetylated proteins using cation-exchange chromatography.
abstract: High-performance liquid chromatography (HPLC) is extremely useful for the study of proteins and the characterization of their posttranslational modifications. Here we describe a method that utilizes cation-exchange HPLC to separate multiply acetylated histone H3 species on the basis of their charge and hydrophilicity. This high-resolution method allows for the separation of histone H3 species that differ by as few as one acetyl group, and is compatible with subsequent analysis by a variety of techniques, including mass spectrometry and western blotting.
authors: Department of Pharmacology and Molecular Sciences, Johns Hopkins University School of Medicine, Baltimore, MD, USA.
authors: Romeo
authors: R
authors: Papazyan
authors: Sean D
authors: SD
authors: Taverna
mesh_terms: D000107:Acetylation; D000818:Animals; D015153:Blotting, Western / Q000379:methods; D002412:Cations / Q000378:metabolism; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D002852:Chromatography, Ion Exchange / Q000379:methods*; D006657:Histones / Q000032:analysis / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism*; D013058:Mass Spectrometry / Q000379:methods; D011499:Protein Processing, Post-Translational; D011506:Proteins / Q000032:analysis / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism*; D015800:Protozoan Proteins / Q000032:analysis / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism; D013768:Tetrahymena / Q000378:metabolism
chemical_list: D002412:Cations; D006657:Histones; D011506:Proteins; D015800:Protozoan Proteins
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