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Application of MassSQUIRM for Quantitative Measurements of Lysine Demethylase Activity
Published on: March 11, 2012
A mechanism-based inactivator for histone demethylase LSD1
Jeffrey C Culhane1, Lawrence M Szewczuk, Xin Liu
1Department of Pharmacology and Molecular Sciences, The Johns Hopkins University School of Medicine, Baltimore, Maryland 21205, USA.
Journal of the American Chemical Society
|April 6, 2006
Summary
Researchers developed a selective inhibitor, compound 1, targeting histone demethylase LSD1 (lysine-specific demethylase 1). This mechanism-based inactivator covalently binds to FAD, offering a new tool for studying histone modifications and chromatin remodeling.
Area of Science:
- Biochemistry
- Molecular Biology
- Epigenetics
Background:
- Histone demethylase LSD1 (lysine-specific demethylase 1) is a key enzyme in epigenetic regulation.
- LSD1 catalyzes the removal of methyl groups from histone H3 lysine 4 (H3K4), influencing gene expression.
- Dysregulation of LSD1 activity is implicated in various diseases, including cancer.
Purpose of the Study:
- To design and synthesize novel peptide-based inhibitors targeting LSD1.
- To characterize the mechanism of inhibition and selectivity of the developed compounds.
- To establish a chemical tool for investigating LSD1 function in histone modification and chromatin remodeling.
Main Methods:
- Design and synthesis of peptide-based inhibitor analogues.
- Enzyme inhibition assays to assess LSD1 activity.
- Time-dependent inactivation studies to determine mechanism of action.
- Mass spectrometry to identify covalent adducts.
- Selectivity profiling against related enzymes like monoamine oxidase B.
Main Results:
- Two peptide-based inhibitor analogues were synthesized, with compound 1 showing potent time-dependent inactivation of LSD1.
- Compound 1 contains a propargylamine group and forms a covalent adduct with the FAD cofactor of LSD1.
- Inhibition by compound 1 was protected by the peptide substrate in a concentration-dependent manner.
- Compound 1 demonstrated high selectivity, with no significant inhibition of monoamine oxidase B observed.
- Mass spectrometry confirmed the covalent interaction between compound 1 and FAD.
Conclusions:
- Compound 1 is a selective, mechanism-based inactivator of histone demethylase LSD1.
- The propargylamine functionality and covalent FAD adduct formation are key to its inhibitory activity.
- Compound 1 serves as a valuable chemical probe for studying the biological roles of LSD1 in histone modification and chromatin remodeling.
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