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
. 2022:2529:43-61.
doi: 10.1007/978-1-0716-2481-4_2.

Detection and Quantification of Histone Methyltransferase Activity In Vitro

Nwamaka J Idigo  1 , Philipp Voigt  2   3
Affiliations

Detection and Quantification of Histone Methyltransferase Activity In Vitro

Nwamaka J Idigo et al. Methods Mol Biol. 2022.

Abstract

Histone methyltransferases (HMTs) catalyze the methylation of lysine and arginine residues in histone as well as nonhistone substrates. In vitro histone methyltransferase assays have been instrumental in identifying HMTs, and they continue to be invaluable tools for the study of these important enzymes, revealing novel substrates and modes of regulation.Here we describe a universal protocol to examine HMT activity in vitro that can be adapted to a range of HMTs, substrates, and experimental objectives. We provide protocols for the detection of activity based on incorporation of 3H-labeled methyl groups from S-adenosylmethionine (SAM), methylation-specific antibodies, and quantification of the reaction product S-adenosylhomocysteine (SAH).

Keywords: Chromatin; Histone methylation; Histone posttranslational modification; Nucleosomes; S-adenosylhomocysteine; S-adenosylmethionine; Transcription.

PubMed Disclaimer

Similar articles

See all similar articles

Cited by

References

    1. Greer EL, Shi Y (2012) Histone methylation: a dynamic mark in health, disease and inheritance. Nat Rev Genet 13(5):343–357 - PubMed - PMC - DOI
    1. Li B, Carey M, Workman JL (2007) The role of chromatin during transcription. Cell 128(4):707–719 - PubMed - DOI
    1. Zhou VW, Goren A, Bernstein BE (2011) Charting histone modifications and the functional organization of mammalian genomes. Nat Rev Genet 12(1):7–18 - PubMed - DOI
    1. Musselman CA, Lalonde M-E, Côté J, Kutateladze TG (2012) Perceiving the epigenetic landscape through histone readers. Nat Struct Mol Biol 19(12):1218–1227 - PubMed - PMC - DOI
    1. Bartke T, Vermeulen M, Xhemalce B, Robson SC, Mann M, Kouzarides T (2010) Nucleosome-interacting proteins regulated by DNA and histone methylation. Cell 143(3):470–484 - PubMed - PMC - DOI

Publication types

MeSH terms

LinkOut - more resources