Reengineering Protease Inhibitors to Disrupt Hsp70 Chaperone Function

Area of Science:

• Biochemistry
• Molecular Biology
• Drug Discovery

Background:

• Heat shock protein 70 (Hsp70) chaperones are crucial for protein homeostasis.
• Bacterial Hsp70, DnaK, is a potential antibacterial target due to its roles in growth and stress response.
• Targeting Hsp70s across species with small molecules is challenging.

Purpose of the Study:

• To investigate the interaction of protease inhibitor scaffolds with bacterial DnaK.
• To design novel peptidomimetics for inhibiting Hsp70 chaperone activity.
• To explore generalizable methods for targeting Hsp70s.

Main Methods:

• Structure-function studies of telaprevir analogs against bacterial DnaK.
• X-ray crystallography to analyze co-complexes of DnaK with inhibitors.
• Synthesis of unnatural peptide sequences based on structural insights.

Main Results:

• Peptidomimetic scaffolds, originally designed for protease inhibition, bind to bacterial DnaK.
• Structural analysis revealed specific ligand-protein interactions within DnaK's substrate binding domain.
• Designed peptidomimetics demonstrated varying inhibitory potencies against DnaK.
• Shortened peptidomimetics were synthesized to allosterically inhibit DnaK's ATPase activity.

Conclusions:

• Protease inhibitor scaffolds can be adapted to inhibit Hsp70 chaperone function.
• This study provides a foundation for developing novel antibacterial agents targeting bacterial Hsp70.
• The findings offer a generalizable strategy for modulating Hsp70 activity across different species.