[Frontiers in Bioscience S1, 142-153, June 1, 2009]

Mass Spectrometric Analysis of Histone Variants and Post-translational Modifications

Benjamin A. Garcia

Department of Molecular Biology, Princeton University, Princeton, NJ

TABLE OF CONTENTS

1. Abstract
2. Introduction
3. Bottom Up mass spectrometry (MS) for histone analysis
3.1. Bottom Up MS identification of histone modification sites
3.2. Chemical derivatization for improved MS analysis
3.3. Determination of histone variants species
3.4. Phosphorylation of histone proteins
4. Comparative MS analysis of histone post-translational modifications (PTMs)
4.1. Label-free quantification of histone PTMs
4.2. Chemical stable isotope labeling of histones for differential expression experiments
4.3. Stable isotope labeling by amino acids in cell culture (SILAC)
5. Top and Middle Down MS for interrogation of intact Histone Codes
5.1. Liquid chromatography MS profiling of modified histone species
5.2. Electron capture dissociation (ECD) of histones
5.3. Electron transfer dissociation (ETD) of histones
6. Conclusions and future perspectives
7. Acknowledgments
8. References

1. ABSTRACT

Mass spectrometry based proteomics has revolutionized many aspects of modern biological research. One key area where mass spectrometry continues to significantly contribute is in the analysis of histone post-translational modification (PTM) patterns. Dynamic histone PTMs are known to be intricately associated with gene regulation (both activating and silencing), and also with epigenetic processes, therefore, accurate qualitative and quantitative mapping of modification sites on these proteins is of immense value. Mass spectrometry has been utilized to confirm, discover, quantify and determine the simultaneous combination of histone PTMs from many organisms. Here the recent mass spectrometry based studies of histone variants and the characterization of their modifications is reviewed.