Employing deep mutational scanning in the <i>Escherichia coli</i> periplasm to decode the thermodynamic landscape for amyloid formation.

ABSTRACT: Deep mutational scanning (DMS) assays provide a powerful method to generate large-scale datasets essential for advancing AI-driven predictions in biology. The tripartite β-lactamase assay (TPBLA), in which a protein of interest is inserted between two domains of β-lactamase, has previously been reported as capable of detecting and quantitating the aggregation of proteins and biologics in the oxidizing periplasm of Escherichia coli and used as a platform for identifying small molecule inhibitors of aggregation. Here, we repurpose the TPBLA into a high-throughput DMS platform. We validate this format using a single-site saturation library of the intrinsically disordered peptide Aβ₄₂, linked to Alzheimer's disease, demonstrating strong agreement between observed variant fitn

SUBMITTER: McKay CE 

PROVIDER: S-EPMC12478104 | biostudies-literature | 2025 Sep

REPOSITORIES: biostudies-literature