A genetically encoded ¹⁹F NMR probe for lysine acetylation

Area of Science:

• Biochemistry
• Molecular Biology
• Chemical Biology

Background:

• Acetylated protein-protein/DNA interactions are crucial in cellular processes.
• Studying conformational changes in acetylated proteins requires advanced methods.
• Current methods for detecting acetylated protein conformation are limited.

Purpose of the Study:

• To develop a novel Nuclear Magnetic Resonance (NMR) probe for studying acetylated protein conformational changes.
• To investigate acetylated p53-protein/DNA interactions using the developed probe.
• To explore the role of SIRT2 in deacetylating p53 K164.

Main Methods:

• Genetic code expansion was used to incorporate an unnatural amino acid.
• A novel 19F NMR probe, mimicking acetyllysine, was synthesized and utilized.
• The probe's interaction with acetylated p53-protein/DNA complexes was analyzed using 19F NMR spectroscopy.
• In vitro and in vivo deacetylation assays were performed.

Main Results:

• A 19F NMR probe with high structural similarity to acetyllysine was successfully developed.
• The probe enabled detection of conformational changes in acetylated p53-protein/DNA interactions.
• The developed probe was resistant to deacetylation by sirtuin deacetylase.
• The deacetylation of p53 K164 by SIRT2 was reported for the first time.

Conclusions:

• The novel 19F NMR probe is a valuable tool for studying acetylated protein dynamics.
• This method advances the understanding of acetylated protein-protein/DNA interactions.
• New insights into the regulation of p53 acetylation by SIRT2 were gained.