Protein mass analysis of histones

Posttranslational modification of chromatin-associated proteins, including histones and high-mobility-group (HMG) proteins, provides an important mechanism to control gene expression, genome integrity, and epigenetic inheritance. Protein mass analysis provides a rapid and unbiased approach to monito...

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Bibliographic Details
Published in:	Methods (San Diego, Calif.) Vol. 31; no. 1; pp. 3 - 11
Main Authors:	Galasinski, Scott C, Resing, Katheryn A, Ahn, Natalie G
Format:	Journal Article
Language:	English
Published:	United States Elsevier Inc 01-09-2003
Subjects:	Acetylation Animals Cell Cycle Centrifugation - methods Chromatin Chromatography, Liquid - methods High Mobility Group Proteins - chemistry High Mobility Group Proteins - metabolism High-mobility-group proteins Histones Histones - chemistry Histones - metabolism Mass spectrometry Methylation Modifications Molecular Weight Phosphorylation Protein Processing, Post-Translational Spectrometry, Mass, Electrospray Ionization - methods Modifications Chromatin Phosphorylation Histones High-mobility-group proteins Acetylation Methylation Mass spectrometry
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Summary:	Posttranslational modification of chromatin-associated proteins, including histones and high-mobility-group (HMG) proteins, provides an important mechanism to control gene expression, genome integrity, and epigenetic inheritance. Protein mass analysis provides a rapid and unbiased approach to monitor multiple chemical modifications on individual molecules. This review describes methods for acid extraction of histones and HMG proteins, followed by separation by reverse-phase chromatography coupled to electrospray ionization mass spectrometry (LC/ESI-MS). Posttranslational modifications are detected by analysis of full-length protein masses. Confirmation of protein identity and modification state is obtained through enzymatic digestion and peptide sequencing by MS/MS. For differentially modified forms of each protein, the measured intensities are semiquantitative and allow determination of relative abundance and stoichiometry. The method simultaneously detects covalent modifications on multiple proteins and provides a facile assay for comparing chromatin modification states between different cell types and/or cellular responses.
Bibliography:	ObjectType-Article-1 SourceType-Scholarly Journals-1 ObjectType-Feature-2 content type line 23
ISSN:	1046-2023 1095-9130
DOI:	10.1016/S1046-2023(03)00082-3