Visualization of Protein‐Specific Glycation in Living Cells via Bioorthogonal Chemical Reporter

Abstract Due to the lack of suitable chemical tools, probing the protein‐specific glycation is highly challenging. Herein, we present a strategy based on glycation chemical reporter and proximity‐induced FRET signal readout for visualizing protein‐specific glycation in living cells. We first developed a bioorthogonal glucose analogue, 6‐azido‐6‐deoxy‐D‐glucose ( 6AzGlc ), as a novel glycation chemical reporter. Two types of DNA probes, glycation conversion probe and protein targeting probe, were designed to attach to glycation adducts and target proteins, respectively. After the protein was glycated by 6AzGlc , two DNA probes were sequentially applied to the target protein, triggering proximity‐induced FRET signal readout. This strategy was successfully used to visualize glucose glycation of several proteins, including PD‐L1 and integrin. More importantly, this strategy allowed us to analyze corresponding biological functions of glycated protein in the native environment.