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Updated: May 19, 2026
Analysis of Histone Antibody Specificity with Peptide Microarrays
Published on: August 1, 2017
Development and validation of reagents and assays for EZH2 peptide and nucleosome high-throughput screens
Elsie Diaz1, Carl A Machutta, Stephanie Chen
1Platform Technology and Science, GlaxoSmithKline, Collegeville, PA 19426, USA.
Abstract:
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 [(3)H]-S-adenosyl-L-methionine competitive, and display biochemical inhibition of H3K27 methylation.
Insights
Researchers identified new inhibitors for EZH2, an enzyme linked to cancer progression. These compounds target histone methylation, offering a potential new avenue for cancer drug development.
Area of Science:
- Biochemistry
- Molecular Biology
- Oncology
Background:
- Histone methyltransferases (HMT) regulate gene transcription through histone methylation.
- Dysregulation of HMTs, like EZH2, is implicated in various cancers.
- EZH2, a key component of PRC2, methylates histone H3 lysine 27 (H3K27) and is overexpressed in multiple tumor types.
Purpose of the Study:
- To develop novel chemotherapeutics targeting EZH2.
- To identify and characterize inhibitors of EZH2 activity.
Main Methods:
- Development of new reagents and assays for EZH2 inhibition.
- Execution of two high-throughput screening campaigns.
- Activity assays using H3K27 peptide or nucleosome substrates.
Main Results:
- Identification of a tractable series of EZH2 inhibitors through screening.
- Compounds demonstrated reversible and competitive inhibition with [(3)H]-S-adenosyl-L-methionine.
- Biochemical inhibition of H3K27 methylation was observed.
Conclusions:
- A novel series of EZH2 inhibitors was discovered.
- These inhibitors show potential for development as anti-cancer therapeutics.
- Targeting EZH2 represents a promising strategy in cancer treatment.
