Site-specific bioorthogonal protein labelling by tetrazine ligation using endogenous β-amino acid dienophiles.

ABSTRACT: The tetrazine ligation is an inverse electron-demand Diels-Alder reaction widely used for bioorthogonal modifications due to its versatility, site specificity and fast reaction kinetics. A major limitation has been the incorporation of dienophiles in biomolecules and organisms, which relies on externally added reagents. Available methods require the incorporation of tetrazine-reactive groups by enzyme-mediated ligations or unnatural amino acid incorporation. Here we report a tetrazine ligation strategy, termed TyrEx (tyramine excision) cycloaddition, permitting autonomous dienophile generation in bacteria. It utilizes a unique aminopyruvate unit introduced by post-translational protein splicing at a short tag. Tetrazine conjugation occurs rapidly with a rate constant of 0.625 (15) M^-1

SUBMITTER: Richter D 

PROVIDER: S-EPMC10533398 | biostudies-literature | 2023 Oct

REPOSITORIES: biostudies-literature