Development and validation of reagents and assays for EZH2 peptide and nucleosome high-throughput screens

Elsie Diaz, Carl A. MacHutta, Stephanie Chen, Yong Jiang, Christopher Nixon, Glenn Hofmann, Danielle Key, Sharon Sweitzer, Mehul Patel, Zining Wu, Caretha L. Creasy, Ryan G. Kruger, Louis Lafrance, Sharad K. Verma, Melissa B. Pappalardi, Baochau Le, Glenn S. Van Aller, Michael T. McCabe, Peter J. Tummino, Andrew J. PopeSara H. Thrall, Benjamin Schwartz, Martin Bandt

Research output: Contribution to journalArticlepeer-review

48 Scopus citations


Histone methyltransferases (HMT) catalyze the methylation of histone tail lysines, resulting in changes in gene transcription. Misregulation of these enzymes has been associated with various forms of cancer, making this target class a potential new area for the development of novel chemotherapeutics. EZH2 is the catalytic component of the polycomb group repressive complex (PRC2), which selectively methylates histone H3 lysine 27 (H3K27). EZH2 is overexpressed in prostate, breast, bladder, brain, and other tumor types and is recognized as a molecular marker for cancer progression and aggressiveness. Several new reagents and assays were developed to aid in the identification of EZH2 inhibitors, and these were used to execute two high-throughput screening campaigns. Activity assays using either an H3K27 peptide or nucleosomes as substrates for methylation are described. The strategy to screen EZH2 with either a surrogate peptide or a natural substrate led to the identification of the same tractable series. Compounds from this series are reversible, are [ 3H]-S-adenosyl-L-methionine competitive, and display biochemical inhibition of H3K27 methylation.

Original languageEnglish (US)
Pages (from-to)1279-1292
Number of pages14
JournalJournal of Biomolecular Screening
Issue number10
StatePublished - Dec 2012
Externally publishedYes


  • EZH2
  • HTS
  • epigenetics
  • histone methyltransferase

ASJC Scopus subject areas

  • Analytical Chemistry
  • Biotechnology
  • Biochemistry
  • Molecular Medicine
  • Pharmacology
  • Drug Discovery


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