Tuning Encodable Tetrazine Chemistry for Site-Specific Protein Bioorthogonal Ligations.

ABSTRACT: Using genetic code expansion (GCE) to encode bioorthogonal chemistry has emerged as a promising method for protein labeling, both in vitro and within cells. Here, we demonstrate that tetrazine (Tet) amino acids incorporated into proteins are highly tunable and have extraordinary potential for fast and quantitative bioorthogonal ligations. We describe the synthesis and characterization of reaction rates for 29 Tet amino acids (20 of which are new) and compare their encoding ability into proteins using evolved tRNA/RS pairs. For these systems, we characterized on-protein Tet stability, reaction rates, and ligation extents as the utility of a bioorthogonal labeling group depends on its stability and reactivity when encoded into proteins. By integrating data on encoding efficiency, selectivity

SUBMITTER: Jana S 

PROVIDER: S-EPMC12662020 | biostudies-literature | 2025 Aug

REPOSITORIES: biostudies-literature