A designed supramolecular protein assembly with in vivo enzymatic activity

• DOI: 10.1126/science.1259680
• Publication Date: 2014-12-19T08:12:06Z
• Authors: Woon Ju Song; F. Akif Tezcan
• Keywords: Protein design

The generation of new enzymatic activities has mainly relied on repurposing the interiors preexisting protein folds because challenge in designing functional, three-dimensional structures from first principles. Here we report an artificial metallo-β-lactamase, constructed via self-assembly a structurally and functionally unrelated, monomeric redox into tetrameric assembly that possesses catalytic zinc sites its interfaces. designed metallo-β-lactamase is functional Escherichia coli periplasm enables bacteria to survive treatment with ampicillin. In vivo screening libraries yielded variant displays proficiency [( k cat / K m )/ uncat ] for ampicillin hydrolysis 2.3 × 10 6 features emergence highly mobile loop near active site, key component natural β-lactamases enable substrate interactions.