A Luminescence-Based System for Identification of Genetically Encodable Inhibitors of Protein Aggregation

[Image: see text] Molecules that disrupt protein aggregation represent potential tool compounds for the investigation of numerous human disease states. However, the identification of small molecules capable of disrupting protein aggregation has proven challenging. Larger biomolecules such as antibodies and proteins are promising alternatives due to their increased size. Despite the promise of protein-based inhibitors, generalizable assays are needed to more readily identify proteins capable of inhibiting aggregation. Herein, we utilize our previously reported self-assembling NanoLuc luciferase fragments to engineer a platform in which both detection reagents are expressed from the same plasmid, enabling facile co-transformation with a genetically encodable inhibitor. This streamlined system is capable of detecting changes in the solubility of amylin, huntingtin, and amyloid-β (Aβ) proteins in response to mutations, small-molecule inhibitors, and expression of genetically encodable inhibitors. This improved platform provides a means to begin to identify protein-based inhibitors with improved efficacy.