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Mol Endocrinol. 2003 Sep;17(9):1704-14. Epub 2003 Jun 19.

Helix 1/8 interactions influence the activity of nuclear receptor ligand-binding domains.

Author information

1
GlaxoSmithKline Research and Development, Research Triangle Park, North Carolina 27709-3398, USA.

Abstract

The ligand-binding domain (LBD) of apo-nuclear receptors in solution is thought to be a very dynamic structure with many possible conformations. Upon ligand binding, the structure is stabilized to a more rigid conformation. The dynamic stabilization assay is a LBD reassembly assay that takes advantage of the high specificity of the intramolecular interactions that comprise the ligand-bound LBD. Here, we demonstrate dynamic stabilization for the nuclear receptors peroxisome proliferator-activated receptor (PPAR)gamma and nerve growth factor inducible (NGFIB)beta and identify residues important for stabilization of the intramolecular interactions induced by PPARgamma ligands. Site-directed mutagenesis studies identified residues in helices 1 and 8 required for LBD reassembly. Further, disrupting the helix 1/8 interaction in the context of the holo-LBD alters the response of the receptor in a compound-specific manner, suggesting that residues far from the ligand-binding pocket can influence the stability of the ligand-bound receptor. Thus, these results support and extend models of the apo-LBD of PPARgamma as a dynamic structure.

PMID:
12817079
DOI:
10.1210/me.2003-0001
[Indexed for MEDLINE]

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