Why do we need a three-dimensional architecture of the ligand-binding domain of the nuclear 1alpha,25-dihydroxyvitamin D(3) receptor?

Highly specific binding of 1alpha,25-dihydroxyvitamin D(3) (1alpha,25(OH)(2)D(3)) by vitamin D receptor (VDR), a nuclear transcriptional factor, activates a genomic mechanism that is manifested in the multiple biologic properties of 1alpha,25(OH)(2)D(3). Numerous synthetic analogs of 1alpha,25(OH)(2)D(3) have been employed to study the interaction between 1alpha,25(OH)(2)D(3) and VDR, and to identify structural markers in 1alpha,25(OH)(2)D(3) that are important for VDR-binding. On the other hand the three-dimensional structure of VDR remained elusive till very recently. In the present study we employed affinity labeling (by 1alpha,25-dihydroxyvitamin D(3)-3-bromoacetate, 1alpha,25(OH)(2)D(3)-3-BE) of VDR to identify C(288) as the anchoring residue for the 3-hydroxyl group of 1alpha,25(OH)(2)D(3) inside the ligand-binding domain of VDR (VDR-LBD). In addition we carried out mutation/hormone-binding analyses to determine the importance of M(284) and W(286) toward hormone binding. We incorporated this information with the three-dimensional structure of the LBD of progesterone receptor to develop a homology-extension model of VDR-LBD. This model identified several amino acid residues as ligand-contact points inside the LBD. Mutational and hormone-binding analyses of these residues verified the structure-functional authenticity of this model, in comparison with the crystal structure of VDR, bound to 1alpha,25(OH)(2)D(3).