Exploiting SpyTag/SpyCatcher Technology to Design New Artificial Catalytic Copper Proteins.

ABSTRACT: Designing artificial metal binding sites within a protein is challenging since amino acid residues need to be placed in desired positions in the final construct and the use of non-natural amino acids is difficult. The alternative approach of directing the insertion of artificial metal coordination systems presents the difficulty of grafting such site in a single desired position. Spy protein is composed of a protein component (SpyCatcher) which binds spontaneously an oligopeptide (SpyTag) with formation of an isopeptide bond. A SpyTag peptide equipped with an ATCUN (amino terminal copper and nickel) binding site is designed to bind copper(II) with high femtomolar affinity both in the absence of SpyCatcher and in the reconstituted Spy construct. The Cu²⁺ ATCUN site in the reconst

SUBMITTER: Gentili S 

PROVIDER: S-EPMC12278347 | biostudies-literature | 2025 Jul

REPOSITORIES: biostudies-literature