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Detection and Quantification of Histone Methyltransferase Activity In Vitro

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Histone Methyltransferases

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2529))

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).

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Acknowledgments

We thank Pablo De Ioannes and Karim-Jean Armache for helpful discussions and advice on SAH quantification methods. Work in the Voigt lab is supported by the Wellcome Trust ([104175/Z/14/Z], Sir Henry Dale Fellowship to P.V.) and through funding from the European Research Council (ERC) under the European Union’s Horizon 2020 research and innovation programme (ERC-STG grant agreement No. 639253 to P.V.). The Wellcome Centre for Cell Biology is supported by core funding from the Wellcome Trust [203149].

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Authors and Affiliations

  1. Epigenetics Programme, Babraham Institute, Cambridge, UK

    Philipp Voigt

  2. Wellcome Centre for Cell Biology, School of Biological Sciences, University of Edinburgh, Edinburgh, UK

    Nwamaka J. Idigo & Philipp Voigt

Authors
  1. Nwamaka J. Idigo
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  2. Philipp Voigt
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Correspondence to Philipp Voigt .

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Editors and Affiliations

  1. Institute Curie, INSERM U934/CNRS UMR 3215, Paris, France

    Raphaël Margueron

  2. Institute Curie, INSERM U934/CNRS UMR 3215, Paris, France

    Daniel Holoch

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Idigo, N.J., Voigt, P. (2022). Detection and Quantification of Histone Methyltransferase Activity In Vitro. In: Margueron, R., Holoch, D. (eds) Histone Methyltransferases. Methods in Molecular Biology, vol 2529. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-2481-4_2

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  • DOI: https://doi.org/10.1007/978-1-0716-2481-4_2

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-2480-7

  • Online ISBN: 978-1-0716-2481-4

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