Site-specific bioorthogonal protein labelling by tetrazine ligation using endogenous β-amino acid dienophiles
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Date
2023-10
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Journal Article
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yes
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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 s−1 and was applied to produce a radiolabel chelator-modified Her2-binding Affibody and intracellular, fluorescently labelled cell division protein FtsZ. We anticipate the labelling strategy to be useful for intracellular studies of proteins, as a stable conjugation method for protein therapeutics, as well as other applications.
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published
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15 (10)
Pages / Article No.
1422 - 1430
Publisher
Nature
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03980 - Piel, Jörn / Piel, Jörn
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742739 - Tailored chemical complexity through evolution-inspired synthetic biology (EC)
ETH-21 21-2 - A single-cell bacterial synthetic biology platform for the biosynthesis and screening of protease-inhibitory peptide drug leads (ETHZ)
ETH-21 21-2 - A single-cell bacterial synthetic biology platform for the biosynthesis and screening of protease-inhibitory peptide drug leads (ETHZ)