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Biochemical Reconstitution of Steroid Receptor•Hsp90 Protein Complexes and Reactivation of Ligand Binding
Published on: September 21, 2011
Backbone dynamics of the glucocorticoid receptor DNA-binding domain
H Berglund1, H Kovács, K Dahlman-Wright
1Center for Structural Biochemistry, Karolinska Institutet, Huddinge, Sweden.
Biochemistry
|December 8, 1992
Summary
This study investigated rapid backbone motions in the glucocorticoid receptor DNA-binding domain (GR DBD) using NMR spectroscopy. The GR DBD exhibits limited and uniform picosecond flexibility, suggesting the second zinc domain is not disordered in its uncomplexed state.
Area of Science:
- Structural Biology
- Biophysics
- Molecular Biology
Background:
- The glucocorticoid receptor DNA-binding domain (GR DBD) plays a crucial role in gene regulation.
- Understanding the dynamic properties of the GR DBD is essential for elucidating its mechanism of action.
- Previous studies have not fully characterized the rapid backbone motions within the GR DBD.
Purpose of the Study:
- To investigate the extent of rapid (picosecond) backbone motions within the GR DBD.
- To determine if the second zinc finger domain is disordered in the uncomplexed state.
Main Methods:
- Proton-detected heteronuclear Nuclear Magnetic Resonance (NMR) spectroscopy was employed on uniformly 15N-labeled GR DBD protein fragments.
- Sequence-specific 15N resonance assignments were obtained using two- and three-dimensional heteronuclear NMR spectra.
- 15N spin-lattice relaxation times (T1), rotating-frame spin-lattice relaxation times (T1 rho), and steady-state (1H)-15N nuclear Overhauser effects (NOEs) were measured.
Main Results:
- 65 of 69 backbone amides within the C440-A509 segment of the rat GR DBD were assigned.
- Analysis of relaxation and NOE data revealed limited and uniform picosecond backbone flexibility across the GR DBD.
- No evidence for extensive rapid backbone motions was found in the second zinc domain.
Conclusions:
- The GR DBD backbone exhibits restricted flexibility on the picosecond timescale.
- The second zinc domain of the GR DBD is likely not disordered in the uncomplexed state.
- Further investigation may be needed to exclude slowly exchanging ordered conformational states.
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