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Chapters
- 00:05Title
- 01:37Preparation of Cell Cytosol Containing Functional Glucocorticoid Receptor
- 03:04Immunoabsorption of GR from the Cytosol and Dissociation of hsp90
- 05:16Reconstitution of GR-hsp90 Heterocomplex using Molecular Chaperones, Cofactors, and ATP
- 06:17Analysis of Reconstituted Protein Complexes and Ligand Binding Activity
- 08:26Representative GR-hsp90 Heterocomplex Formation and Steroid Binding Activity
- 10:04Conclusion
Um<em> In vitro</em> Método para preparar receptor glicocorticóide funcional (GR) • hsp90 complexos de proteína purificada a partir de proteínas e lisados celulares é descrito. O método utiliza de imunoadsorção recombinante GR seguido de sal de extracção e reconstituição de proteínas complexas. A importância da co-fatores e condições de buffer são discutidos, assim como as aplicações método potencial.
Tags
Biochemical ReconstitutionSteroid ReceptorHsp90 Protein ComplexesLigand BindingMolecular ChaperoneEukaryotic Signaling ProteinsTranscription FactorsProtein KinasesCell CyclingTumorigenesisApoptosisSignaling PathwaysGlucocorticoid Receptor (GR)Immunoprecipitation AssayIn Vitro Reconstitution MethodHsp90 Functional ActivitySteroid Receptor Ligand BindingMolecular Chaperone Cofactor Requirements
