Differential scanning fluorimetry (DSF) screen to identify inhibitors of Hsp60 protein-protein interactions

• Published in: Organic & biomolecular chemistry Vol. 18; no. 22; pp. 4157 - 4163
• Main Authors: Shao, Hao, Oltion, Keely, Wu, Taia, Gestwicki, Jason E
• Format: Journal Article
• Language: English
• Published: England Royal Society of Chemistry 10.06.2020
• Subjects: Fluorimetry; Hsp60 protein; Inhibitors; Monomers; Oligomerization; Oligomers; Protein interaction; Proteins; Scanning; Thermal stability
• ISSN: 1477-0520; 1477-0539
• DOI: 10.1039/d0ob00928h

There are relatively few methods available for discovering inhibitors of the protein-protein interactions (PPIs) that hold together homo-oligomers. We envisioned that Differential Scanning Fluorimetry (DSF) might be a versatile way to discover this type of inhibitor because oligomers are often more thermally stable than monomers. Using the homo-heptameric chaperonin, Hsp60, as a model, we screened ∼5000 diverse compounds in 384-well plates by DSF, revealing molecules that partially inhibited oligomerization. Because DSF does not require protein labeling or structural information, we propose that it could be a versatile way to uncover PPI inhibitors. Homo-oligomers are difficult drug targets. Here, Differential Scanning Fluorimetry (DSF) is introduced as a method to identify inhibitors of these systems, by discriminating between oligomers and monomers based on their thermal stability.