Importance

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Abstract

Described are novel ways of introducing an affinity tag into a protein of interest. Provided is an enzymatic method for providing a proteinaceous substance comprising a polypeptide of interest and a cyclic affinity tag, comprising: (a) providing at least one precursor proteinaceous substance, the precursor comprising the protein of interest and at least one motif of the general formula X1-Tag-X2, wherein X1 and X2 represent amino acids whose side chains can be linked enzymatically by a covalent bond; Tag is a short amino acid sequence capable of binding to a binding partner of the tag when cyclized; (b) contacting the precursor with at least one enzyme capable of forming a covalent bond between X1 and X2, thereby introducing an intramolecular ring structure comprising the Tag sequence; and (c) isolating the resulting cyclized proteinaceous substance. Also provided are proteinaceous substances obtainable thereby and the use thereof, for instance, for preparing a peptide library.

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First Claim