Nonmodified Strategy Enabled Proteome-Wide Mapping of Catechol Derivatives Interactome

Area of Science:

• Chemical Biology
• Pharmacology
• Proteomics

Background:

• Understanding catechol derivative pharmacology requires profiling their protein interactions.
• Current methods for target identification often involve ligand modification or protein perturbation, limiting comprehensive interactome coverage.

Purpose of the Study:

• To develop a modification-free strategy for mapping catechol-protein interactions in native cellular environments.
• To enable direct analysis of the interactome of catechol-containing compounds.

Main Methods:

• Developed oxidative addition-boronate affinity enrichment (OABA), a redox-enabled technique.
• Utilized catechol oxidation to ortho-quinones for covalent protein capture.
• Employed boronate affinity recognition for selective enrichment of catechol-protein adducts.

Main Results:

• Demonstrated proof-of-concept with isoproterenol (ISO), achieving 2-fold enrichment of SOD1-adduct peptides.
• Identified 74 high-confidence quercetin interactors in live HeLa cells.
• Validated interactions with FDPS, CCT7, and CS using molecular docking and CETSA.

Conclusions:

• OABA provides a reliable, modification-free platform for mapping native catechol-protein interactomes.
• This method advances the understanding of polyphenolic pharmacology.
• Facilitates direct mapping of drug-protein interactions in living systems.