Proximity-induced protein deglycosylation by endogenous O-GlcNAcase

O-GlcNAcylation, a crucial post-translational modification, influences various cellular functions; however, methods to selectively remove O-GlcNAc from specific proteins using endogenous O-GlcNAcase (OGA) in living cells are scarce. In this study, researchers introduce De-O GlcNAcylation-targeting chimeras (DOGTACs), a proximity strategy that selectively reduces O-GlcNAc from target proteins by leveraging endogenous OGA.

Although initial designs with powerful OGA inhibitors successfully engaged the enzyme, they were unable to induce de-O-GlcNAcylation, highlighting the importance of catalytic activity for successful proximity-driven editing. To address this issue, the team attenuated inhibitor potency while maintaining adequate OGA engagement, resulting in optimized DOGTACs.

These new DOGTACs enable concentration- and time-dependent, target-specific de-O-GlcNAcylation in living cells without disrupting overall O-GlcNAc levels. Furthermore, the researchers successfully applied DOGTAC to additional target proteins across multiple cell lines. Overall, this work establishes attenuated competitive inhibitors as effective tools for recruiting catalytic enzymes and introduces a novel framework, DOGTAC, for targeted de-O-GlcNAcylation via endogenous OGA recruitment in living cells.