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Studies of Chaperone-Cochaperone Interactions using Homogenous Bead-Based Assay
Journal JoVE
Biochimie
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Journal JoVE Biochimie
Studies of Chaperone-Cochaperone Interactions using Homogenous Bead-Based Assay
DOI:

06:51 min

July 21, 2021

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Chapitres

  • 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

Summary

Traduction automatique

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.

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