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Molecular structure of the rat vitamin D receptor ligand binding domain complexed with 2-carbon-substituted vitamin D3 hormone analogues and a LXXLL-containing coactivator peptide

Vanhooke, J.L.; Benning, M.M.; Bauer, C.B.; Pike, J.Wesley.; DeLuca, H.F.

Biochemistry 43(14): 4101-4110

2004


ISSN/ISBN: 0006-2960
PMID: 15065852
DOI: 10.1021/bi036056y
Accession: 009912511

We have determined the crystal structures of the ligand binding domain (LBD) of the rat vitamin D receptor in ternary complexes with a synthetic LXXLL-containing peptide and the following four ligands: 1a,25-dihydroxyvitamin D3; 2-methylene-19-nor-(20S)-1a,25-dihydroxyvitamin D3 (2MD); 1a-hydroxy-2-methylene-19-nor-(20S)-bishomopregnacalciferol (2MbisP), and 2a-methyl-19-nor-1a,25-dihydroxyvitamin D3 (2AM20R). The conformation of the LBD is identical in each complex. Binding of the 2-carbon-modified analogues does not change the positions of the amino acids in the ligand binding site and has no effect on the interactions in the coactivator binding pocket. The CD ring of the superpotent analogue, 2MD, is tilted within the binding site relative to the other ligands in this study and to (20S)-1a,25-dihydroxyvitamin D3 [Tocchini-Valentini et al. (2001) Proc. Natl. Acad. Sci. U.S.A. 98, 5491 - 5496]. The aliphatic side chain of 2MD follows a different path within the binding site; nevertheless, the 25-hydroxyl group at the end of the chain occupies the same position as that of the natural ligand, and the hydrogen bonds with histidines 301 and 393 are maintained. 2MbisP binds to the receptor despite the absence of the 25-hydroxyl group. A water molecule is observed between His 301 and His 393 in this structure, and it preserves the orientation of the histidines in the binding site. Although the a-chair conformer is highly favored in solution for the A ring of 2AM20R, the crystal structures demonstrate that this ring assumes the b-chair conformation in all cases, and the 1a-hydroxyl group is equatorial. The peptide folds as a helix and is anchored through hydrogen bonds to a surface groove formed by helices 3, 4, and 12. Electrostatic and hydrophobic interactions between the peptide and the LBD stabilize the active receptor conformation. This stablization appears necessary for crystal growth. Reprinted by permission of the publisher.

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