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Published on: February 27, 2016
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An improved fluorogenic assay for SIRT1, SIRT2, and SIRT3
1Department of Chemistry and Chemical Biology, Baker Laboratory, Cornell University, Ithaca, NY 14853, USA.
Organic & Biomolecular Chemistry
|January 23, 2016
Summary
Researchers developed a new, sensitive assay to detect sirtuin activity. This improved fluorogenic assay enables faster screening for sirtuin modulators, crucial for understanding cellular regulation and disease.
Area of Science:
- Biochemistry
- Molecular Biology
- Enzymology
Background:
- Sirtuins are critical NAD-dependent lysine deacylases involved in cellular regulation.
- Dysregulation of sirtuins is linked to various human diseases.
- Efficient assays are essential for discovering sirtuin modulators for research and therapeutics.
Purpose of the Study:
- To develop an improved fluorogenic assay for sirtuin enzymes, specifically SIRT1, SIRT2, and SIRT3.
- To facilitate the discovery of novel sirtuin modulators.
Main Methods:
- A novel fluorogenic substrate, myristoyl peptide with a C-terminal aminocoumarin, was synthesized.
- The substrate was utilized in a new assay designed for high-throughput screening of sirtuin activity.
Main Results:
- The new assay requires significantly lower substrate concentrations.
- It demonstrates an increased signal-to-background ratio.
- An improved Z'-factor was achieved, indicating assay robustness.
Conclusions:
- The developed fluorogenic assay is highly efficient for SIRT1, SIRT2, and SIRT3.
- This assay will accelerate the high-throughput screening of sirtuin modulators.
- It provides a valuable tool for sirtuin research and drug discovery.
