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

Authors

Travis J. Nelson, University of Nebraska - Lincoln
Shuo Liang, University of Nebraska - Lincoln
Cliff I. Stains, University of Nebraska - Lincoln & University of Nebraska Medical Center & University of VirginiaFollow

ORCID IDs

http://orcid.org/0000-0002-3165-4539

Date of this Version

2020

Citation

ACS Omega 2020, 5, 12974−12978

Comments

2020 American Chemical Society

Abstract

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.