Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The .gov means it’s official.
Federal government websites often end in .gov or .mil. Before sharing sensitive information, make sure you’re on a federal government site.

Https

The site is secure.
The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely.

Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
. 2023:2718:271-283.
doi: 10.1007/978-1-0716-3457-8_15.

Mass Spectrometry-Based Analysis of Histone Posttranslational Modifications from Laser Microdissected Samples

Roberta Noberini  1 , Tiziana Bonaldi  2   3
Affiliations

Mass Spectrometry-Based Analysis of Histone Posttranslational Modifications from Laser Microdissected Samples

Roberta Noberini et al. Methods Mol Biol. 2023.

Abstract

The analysis of histone posttranslational modifications (PTMs) in clinical samples has gained considerable interest due to the increasing knowledge about the implication of epigenetics in a multitude of physiological and pathological processes. Mass spectrometry (MS) has emerged as the most accurate and versatile tool to detect and quantify histone PTMs and has also been applied to clinical specimens, thanks to protocols developed during the past years. However, the requirement for relatively large amounts of material has so far impaired the application of these approaches to samples available in limited amounts. To address this issue, we have recently streamlined the protein extraction procedure from low-amount clinical samples and optimized the digestion step, obtaining a protocol suitable for the analysis of the most common histone PTMs from laser microdissected tissue areas containing down to 1000 cells, which we will describe in this chapter.

Keywords: Epigenetics; Histone posttranslational modifications; Laser microdissection; Mass spectrometry.

PubMed Disclaimer

References

    1. Jenuwein T, Allis CD (2001) Translating the histone code. Science 293(5532):1074–1080. https://doi.org/10.1126/science.1063127 . 293/5532/1074 [pii] - DOI - PubMed
    1. Portela A, Esteller M (2010) Epigenetic modifications and human disease. Nat Biotechnol 28(10):1057–1068. https://doi.org/10.1038/nbt.1685 . nbt.1685 [pii] - DOI - PubMed
    1. Lu C, Coradin M, Porter EG, Garcia BA (2021) Accelerating the field of epigenetic histone modification through mass spectrometry-based approaches. Mol Cell Proteomics 20:100006. https://doi.org/10.1074/mcp.R120.002257 - DOI - PubMed
    1. Noberini R, Longuespee R, Richichi C, Pruneri G, Kriegsmann M, Pelicci G, Bonaldi T (2017) PAT-H-MS coupled with laser microdissection to study histone post-translational modifications in selected cell populations from pathology samples. Clin Epigenetics 9:69. https://doi.org/10.1186/s13148-017-0369-8 . 369 [pii] - DOI - PubMed - PMC
    1. Noberini R, Osti D, Miccolo C, Richichi C, Lupia M, Corleone G, Hong SP, Colombo P, Pollo B, Fornasari L, Pruneri G, Magnani L, Cavallaro U, Chiocca S, Minucci S, Pelicci G, Bonaldi T (2018) Extensive and systematic rewiring of histone post-translational modifications in cancer model systems. Nucleic Acids Res 46:3817. https://doi.org/10.1093/nar/gky224 . 4956179 [pii] - DOI - PubMed - PMC

Publication types

LinkOut - more resources