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Biochemical Reconstitution of Steroid Receptor•Hsp90 Protein Complexes and Reactivation of Ligand Binding
 

Biochemical Reconstitution of Steroid Receptor•Hsp90 Protein Complexes and Reactivation of Ligand Binding

Article DOI: 10.3791/3059-v • 11:08 min • September 21st, 2011
September 21st, 2011 •
1, 2, 3
1College of Nursing, Interdisciplinary Life Sciences Research Laboratory, Seattle University, 2College of Science and Engineering, Interdisciplinary Life Sciences Research Laboratory, Seattle University, 3School of Medicine, University of Washington

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An in vitro method for preparing functional glucocorticoid receptor (GR)•hsp90 protein complexes from purified proteins and cellular lysates is described. The method utilizes immunoadsorption of recombinant GR followed by salt-stripping and protein complex reconstitution. The importance of cofactors and buffer conditions are discussed, as are potential method applications.

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Biochemical Reconstitution Steroid Receptor Hsp90 Protein Complexes Ligand Binding Molecular Chaperone Eukaryotic Signaling Proteins Transcription Factors Protein Kinases Cell Cycling Tumorigenesis Apoptosis Signaling Pathways Glucocorticoid Receptor (GR) Immunoprecipitation Assay In Vitro Reconstitution Method Hsp90 Functional Activity Steroid Receptor Ligand Binding Molecular Chaperone Cofactor Requirements
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