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JoVE Journal Biochemistry
Studies of Chaperone-Cochaperone Interactions using Homogenous Bead-Based Assay
 

Studies of Chaperone-Cochaperone Interactions using Homogenous Bead-Based Assay

Article DOI: 10.3791/62762-v • 06:51 min • July 21st, 2021
July 21st, 2021 •
1, 1, 1,2, 1,3, 1
1Department of Neuroscience Care and Society, Division of Neurogeriatrics, Karolinska Institutet, 2Department of Pharmaceutical Engineering & Technology, Indian Institute of Technology (BHU), 3Theme Inflammation and Aging, Karolinska University Hospital

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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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Chaperone-cochaperone Interactions Bead-based Assay Hsp90 FKDP51 FKBP52 Small Molecule Inhibitors High Throughput Screening Technique Alzheimer's Disease Cancer Autoimmune Disease Chaperone-cochaperone Inhibitors Hsp90C-Terminal Peptide TPR Domain Proximity-based Signal Amplification Donor Beads Acceptor Beads Interaction Inhibitors PBS Dilution
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