Harnessing Ionic Selectivity in Acetyltransferase Chemoproteomic Probes

Area of Science:

• Chemical proteomics
• Biochemistry
• Drug discovery

Background:

• Chemical proteomics is vital for enzyme function and inhibitor selectivity studies.
• Optimizing chemical probes and reducing background noise are key challenges.
• Coenzyme A (CoA) is a crucial cofactor in many enzymatic reactions.

Purpose of the Study:

• To develop a physiochemical discernment strategy for optimizing chemical proteomics probes.
• To differentiate probe capture properties based on subtle chemical modifications.
• To evaluate the selectivity of small molecule N-terminal acetyltransferase inhibitors.

Main Methods:

• Synthesis of CoA-based sepharose pulldown resins with distinct charge properties.
• Integration of probes with quantitative proteomics and gel-based profiling.
• Benchmarking probe selectivity analysis against traditional competitive chemical proteomics.

Main Results:

• A single charged residue difference significantly altered resin capture properties.
• Probe selectivity analysis is effective for identifying optimized pulldown probes.
• Anionic CoA probes successfully evaluated N-terminal acetyltransferase inhibitor selectivity.

Conclusions:

• Physicochemical discriminant strategies enhance chemoproteomic hit identification.
• CoA-based probes are valuable for assessing small molecule inhibitor selectivity.
• This approach aids in the development of novel therapeutic agents targeting protein acetyltransferases.