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Comparative studies of thiol-sensitive fluorogenic probes for HAT assays
Tielong Gao1, Chao Yang, Yujun George Zheng
1Department of Chemistry, Georgia State University, Atlanta, GA 30302, USA.
Analytical and Bioanalytical Chemistry
|November 10, 2012
Summary
Researchers developed a new fluorescent assay to measure histone acetyltransferase (HAT) activity. This method enables efficient screening for HAT inhibitors, aiding drug discovery.
Area of Science:
- Biochemistry
- Enzymology
- Molecular Biology
Background:
- Histone acetyltransferases (HATs) regulate cellular pathways through protein acetylation.
- Existing HAT activity assays are limited, hindering research and drug discovery.
- Acetylation is a widespread post-translational modification with diverse biological roles.
Purpose of the Study:
- To evaluate thiol-sensitive fluorogenic compounds as novel HAT activity reporters.
- To compare the performance of different fluorogenic probes for HAT detection.
- To establish a robust assay for high-throughput screening of HAT inhibitors.
Main Methods:
- Conjugation of fluorogenic compounds to HSCoA to enhance fluorescence.
- Kinetic analysis of probe reactions with HSCoA.
- Assessment of probe influence on HAT activity and fluorescence amplification.
- Microtiter plate-based HAT activity measurements.
Main Results:
- Fluorogenic compounds, particularly CPM and coumarin maleic acid ester, showed excellent performance.
- These probes exhibit fast reaction kinetics and significant fluorescence enhancement.
- The developed fluorescent assay is robust and suitable for microtiter plate formats.
Conclusions:
- Novel fluorogenic probes enable sensitive and efficient detection of HAT activity.
- This assay strategy is valuable for high-throughput screening of HAT inhibitors.
- The findings support the development of new therapeutic agents targeting HATs.
