Nonmodified Strategy Enabled Proteome-Wide Mapping of Catechol Derivatives Interactome

Profiling the interactome of catechol derivatives plays a key role in revealing their pharmacology. Existing approaches for target identification generally rely on either the modification of ligands or significant perturbation of protein properties, which may restrict the coverage of interacting proteins. Here, we present a modification-free strategy, termed oxidative addition-boronate affinity enrichment (OABA), that employed catechol oxidation to ortho-quinones to covalently capture nucleophilic residues of proteins, followed by selective enrichment via boronate affinity recognition of the catechol moiety. This redox-enabled approach eliminated the requirement for drug molecule modification while enabling direct mapping of catechol-protein interactomes in living cells. Proof-of-concept studies with isoproterenol (ISO) achieved a 2-fold enrichment of SOD1-adduct peptides from the HeLa peptide mixture. Application to quercetin in live HeLa cells identified 74 high-confidence interacting targets, enriched in biological process linked to the modulation of protein stabilization, energy metabolism, and cell growth. Through molecular docking and CETSA profiling, we validated the interaction of FDPS, CCT7, and CS with quercetin. Collectively, the OABA provided a reliable, modification-free platform for mapping the native interactome of catechol-containing compounds and advancing the understanding of polyphenolic pharmacology.