title: Middle-Down and Top-Down Mass Spectrometric Analysis of Co-occurring Histone Modifications.
abstract: Histones are chromatin proteins that are highly modified with many different types of post-translational modifications. These modifications act in concert to regulate a number of chromatin-related processes. However, identification and quantification of co-occurring histone post-translational modifications is challenging because there are many potential combinations of modifications and because the commonly used strategy of fragmenting proteins using trypsin or an alternative protease prior to LC-MS/MS analysis results in the loss of connectivity between modifications on different peptides. In this unit, mass spectrometric methods to analyze combinatorial histone modifications on histone tails (middle-down mass spectrometry) and on intact histones (top-down mass spectrometry) are described.
authors.affiliation: Department of Chemistry, Frick Laboratory, Princeton University, Princeton, New Jersey.
authors.forename: Rosalynn C
authors.initials: RC
authors.lastname: Molden
authors.affiliation: Epigenetics Program, Department of Biochemistry and Biophysics, Perelman School of Medicine, University of Pennsylvania, Philadelphia, Pennsylvania.
authors.forename: Benjamin A
authors.initials: BA
authors.lastname: Garcia
mesh_terms: D002853:Chromatography, Liquid / Q000379:methods; D006657:Histones / Q000032:analysis* / Q000737:chemistry / Q000378:metabolism; D013058:Mass Spectrometry / Q000379:methods*; D010455:Peptides / Q000032:analysis* / Q000737:chemistry / Q000378:metabolism; D011499:Protein Processing, Post-Translational; D014357:Trypsin / Q000737:chemistry*
chemical_list: D006657:Histones; D010455:Peptides; D014357:Trypsin
keywords: histone modifications; middle-down mass spectrometry; post-translational modifications; top-down mass spectrometry
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