New insights into receptor ligand binding domains from a novel assembly assay.
Previous studies have demonstrated that hormone binding stabilizes the ligand binding domain (LBD) of the nuclear hormone receptors against proteolysis. We have confirmed and extended this observation using a newly developed assembly assay. In this assay, the LBD is divided into two parts, of which one includes the first helix of this domain and the other corresponds to the remainder of the LBD. Several independent criteria demonstrate that these two fragments can assemble into a functional LBD in the presence of a ligand, but not in its absence, and that this is a reflection of the stabilizing effect of ligand. We have also used this assay to demonstrate that binding of the nuclear receptor corepressor NCoR can directly stabilize the LBD. Overall, these results highlight the dynamic nature of the LBD and suggest that current models for activation based solely on allosteric effects on the C-terminal helix may be too limited.
Pissios
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Department of Molecular and Cellular Biology, Baylor College of Medicine, One Baylor Plaza, 1200 Moursund Avenue, Houston, TX 77030, USA.
Tzameli
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Moore
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D006868:Hydrolysis; D008024:Ligands; D011485:Protein Binding; D018160:Receptors, Cytoplasmic and Nuclear / Q000378:metabolism*
D008024:Ligands; D018160:Receptors, Cytoplasmic and Nuclear