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Updated: Aug 10, 2025
11:08
A Facile Protocol to Generate Site-Specifically Acetylated Proteins in Escherichia Coli
Published on: December 9, 2017
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A heterobifunctional molecule system for targeted protein acetylation in cells
Li-Yun Chen1, Wesley Wei Wang1, Jacob M Wozniak1
1Department of Chemistry, The Scripps Research Institute, La Jolla, CA, United States.
Methods in Enzymology
|February 10, 2023
Summary
Researchers developed AceTAG, a novel system for chemically inducing protein acetylation in live cells. This tool enables targeted, rapid, and controlled investigations into the roles of protein acetylation in various cellular processes.
Area of Science:
- Molecular Biology
- Chemical Biology
- Proteomics
Background:
- Protein acetylation is a critical post-translational modification regulating diverse cellular functions.
- Existing methods for studying protein acetylation lack dynamic and selective control in live cells.
Purpose of the Study:
- To develop a novel chemical biology tool, AceTAG, for targeted induction of protein acetylation.
- To enable dynamic and selective regulation of protein acetylation in live cells for functional studies.
Main Methods:
- Development of AceTAG, a heterobifunctional molecule linking p300/CBP lysine acetyltransferase to FKBP12-tagged target proteins.
- Genetic tagging of target proteins with FKBP12F36V.
- In vitro and in situ characterization using ternary complex formation assays, immunoblotting, and quantitative proteomics.
Main Results:
- AceTAG successfully induces targeted protein acetylation in live cells.
- The acetylation process is selective, rapid, and dose-dependent.
- Demonstrated robust procedures for AceTAG construct generation, cell line development, and characterization.
Conclusions:
- AceTAG provides a powerful and versatile tool for investigating the functional roles of protein acetylation.
- This system facilitates controlled exploration of acetylated protein targets in cellular contexts.
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