Abstract
We report here a photosensitized strategy for protein labelling in which N-substituted pyridinium salts are activated using a 2,4-diaryl-N-methyl quinolinium scaffold. Structure-reactivity relationships were performed to optimize the sensitizer structure, and ultimately generated a system that gives protein labelling in minutes at micromolar reagent concentrations. Mechanistic studies suggest a photo-induced electron transfer-based sensitization mechanism. The mildness of this system enabled us to assay sensitization both on individual biomolecules and in complex proteomes and demonstrated excellent compatibility with lysate- and the live-cell-based systems. Imaging of photolabelled HeLa cells were performed and revealed that catalysis occurs in multiple cellular compartments. Chemical proteomics performed at the lysate level resulted in the enrichment of 319 proteins with a 93% selectivity to Tryptophan residues. Live cell labelling resulted in 101 enriched proteins, primarily from the nucleus.