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Updated: Jun 23, 2026
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Assays for Validating Histone Acetyltransferase Inhibitors
Published on: August 6, 2020
In vitro histone demethylase assays
1Molecular Oncology Program, H. Lee Moffitt Cancer Center and Research Institute, Tampa, FL, USA.
Methods in Molecular Biology (Clifton, N.J.)
|April 22, 2009
Summary
Histone demethylases dynamically regulate gene expression by removing methylation marks. This study details assay methods and substrate preparation for understanding these crucial epigenetic enzymes.
Area of Science:
- Epigenetics and Molecular Biology
- Enzymology
- Gene Regulation
Background:
- Histone methylation is a key epigenetic mark influencing chromatin structure and transcription.
- Histone methylation is dynamically regulated by histone methyltransferases and demethylases.
- Two main families of histone demethylases, LSD1 and JmjC proteins, utilize distinct cofactors (FAD or Fe(II)/alpha-ketoglutarate) to remove methyl groups.
Purpose of the Study:
- To describe assay conditions and detection methods for different histone demethylase families.
- To provide protocols for the purification and preparation of histone substrates for demethylase assays.
- To advance the understanding of chromatin dynamics and epigenetic regulation.
Main Methods:
- Detailed description of various assay conditions and detection methodologies for histone demethylases.
- Step-by-step protocols for the purification of histone demethylase enzymes.
- Protocols for the preparation of diverse histone substrates essential for enzymatic assays.
Main Results:
- Established diverse assay conditions and detection methods tailored to specific histone demethylase families.
- Provided comprehensive protocols for histone substrate preparation and enzyme purification.
- Facilitated further research into the functional roles and regulatory mechanisms of histone demethylases.
Conclusions:
- The characterization of histone demethylase assays and substrate preparation is crucial for advancing epigenetic research.
- Understanding these enzymes offers insights into fundamental biological processes and human diseases.
- This work provides essential tools for studying the dynamic regulation of chromatin.
