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Updated: Jul 6, 2026
Histone Modification Screening using Liquid Chromatography, Trapped Ion Mobility Spectrometry, and Time-Of-Flight Mass Spectrometry
Published on: January 12, 2024
Proteomic Analysis of Histone Sequence Variants and Post-translationally Modified Forms
Palina Ryzhaya1,2, Pavlína Pírek1, Radomír Pech1
1Mendel Center for Plant Genomics and Proteomics, Central European Institute of Technology, Masaryk University, Brno, Czech Republic.
Abstract:
Post-translational modifications (PTMs) of histone variants are the key mechanisms driving epigenetic regulation. They play a crucial role in modulating chromatin structure and gene activity. As epigenetics has become a foundational aspect of molecular biology, the precise characterization of histones is essential for understanding gene regulation, cellular plasticity, and the molecular basis of diseases.Mass spectrometry in combination with liquid chromatography has emerged as the leading technology for mapping and quantification of histone peptidoforms. It offers exceptional sensitivity and resolution to unravel the complexity of the epigenetic landscape. Despite its advantages, the inherent diversity of histone isoforms and PTMs presents analytical challenges, and no universal workflow currently enables comprehensive and unbiased profiling of the overall state of histone modifications.This chapter presents key methodologies for histone analysis using bottom-up proteomics, including protocols for histone extraction, enzymatic digestion, and chemical derivatization prior to LC-MS/MS. It also provides details of specific aspects of LC-MS/MS analysis and outlines strategies for data evaluation and interpretation. The clinical potential of histone proteomics is illustrated through a case study focusing on the characterization of histones in human sperm in the context of male infertility.
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