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Studies of Chaperone-Cochaperone Interactions using Homogenous Bead-Based Assay
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Chapters
- 00:04Introduction
- 01:28Coupling of Hsp90 C-Terminal Peptide to the Acceptor Beads
- 02:49Assay for Interaction Between GST-FKBP51 or GST-FKBP52 and Hsp90 C-Terminal Peptide and Inhibition with Small Molecular Mass Compounds
- 04:14Dose-Response Curve Generation and IC50 Calculation
- 04:58Results: Inhibitory Effect of Small Molecular Weight Test Compound D10
- 05:59Conclusion
This protocol presents a technique for probing protein-protein interactions using glutathione-linked donor beads with GST-fused TPR-motif co-chaperones and acceptor beads coupled with an Hsp90-derived peptide. We have used this technique to screen small molecules to disrupt Hsp90-FKBP51 or Hsp90-FKBP52 interactions and identified potent and selective Hsp90-FKBP51 interaction inhibitors.
