Site-specific thiopyrophosphorylation and functionalization of phosphate residues in oligonucleotides and peptides

A new strategy is developed for the functionalization of phosphate moieties in oligonucleotides and peptides. Diamidothiophosphate (DATP) acts as a phosphorylating agent to site-specifically attach a thiophosphate group onto oligonucleotide terminal phosphates and phosphate residues in peptides. This thiophosphate structure serves as a highly reactive, bioorthogonal handle for diverse downstream functionalization. Chemoselective functionalization of the thiophosphate moiety is achieved in a mildly acidic aqueous solution, while native thiols are substantially less reactive under the same conditions. This work provides a versatile strategy for functionalizing phosphate groups in biomacromolecules.