Single electron transfer between sulfonium and tryptophan enables site-selective photo crosslinking of methyllysine reader proteins

The identification of readers, an important class of proteins that recognize modified residues at specific sites, is essential to uncover biological roles of posttranslational modifications. Photoreactive crosslinkers are powerful tools for investigating readers. However, existing methods usually employ synthetically challenging photoreactive warheads and their high-energy intermediates generated upon irradiation, such as nitrene and carbene, may cause significant non-specific crosslinking. Here we report dimethylsulfonium as a methyllysine mimic that binds to specific readers and subsequently crosslinks to a conserved tryptophan inside the binding pocket through single electron transfer under ultraviolet irradiation. The crosslinking relies on a protein-templated σ-π electron-donor-acceptor interaction between sulfonium and indole, ensuring excellent site-selectivity for tryptophan in the active site and orthogonality to other methyllysine readers. This method could escalate the discovery of methyllysine readers from complex cell samples. Furthermore, this photo crosslinking strategy could be extended to develop other types of microenvironment-dependent conjugations to site-specific tryptophan.