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Issue 5, 2010
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A chemiluminescence-based method for identification of histone lysine methyltransferase inhibitors

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Abstract

Methylation of lysine residues, catalyzed by histone methyltransferase (HMT) enzymes, is one of many modifications of core histone proteins that regulate transcription and chromatin structure. G9a is the predominant HMT in mammalian euchromatin and recent data suggest that it is required to perpetuate a malignant phenotype in cancer cells and is implicated in metastasis, supporting this HMT as a therapeutic target for cancer and other diseases associated with epigenetic regulation. Of the methods currently used to measure methyltransferase activity, many involve a separation step or utilize coupling enzymes complicating implementation and data interpretation. Here we describe a homogeneous assay to measure G9a HMT activity using the chemiluminescence-based AlphaScreen immunoassay technology. Methylation of biotinylated-histone peptide is measured through specific antibody-based detection, in conjunction with streptavidin-coated donor and secondary antibody-coated acceptor beads. The method is particularly well suited for detection of inhibitors acting by the desired histone peptide competitive mechanism and is applicable to testing other HMTs, demonstrated here with the G9a homolog EHMT1, also known as GLP.

Graphical abstract: A chemiluminescence-based method for identification of histone lysine methyltransferase inhibitors

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Publication details

The article was received on 21 Oct 2009, accepted on 28 Jan 2010 and first published on 02 Mar 2010


Article type: Method
DOI: 10.1039/B921912A
Citation: Mol. BioSyst., 2010,6, 782-788
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    A chemiluminescence-based method for identification of histone lysine methyltransferase inhibitors

    A. M. Quinn, A. Allali-Hassani, M. Vedadi and A. Simeonov, Mol. BioSyst., 2010, 6, 782
    DOI: 10.1039/B921912A

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