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The impact of functional vitamin D(3) receptor conformations on DNA-dependent vitamin D(3) signaling.
Mol Pharmacol 2000; 57(2):375-84MP

Abstract

The vitamin D(3) receptor (VDR) is the nuclear receptor for 1alpha, 25-dihydroxyvitamin D(3) (VD) that acts primarily as a heterodimer with the retinoid X receptor (RXR) on different types of VD response elements, i.e., DNA-bound VDR-RXR heterodimers are the molecular switches in nuclear VD signaling pathways. In this study, DNA-dependent limited protease digestion assays and gel shift clipping assays were used for the analysis of VDR conformations and showed the same high ligand sensitivity for VD response element-bound VDR-RXR heterodimers (EC(50) of 0.1 nM for VD). In contrast, DNA-independent limited protease digestion assays clearly demonstrated a reduced ligand sensitivity for monomeric VDR in solution. Interestingly, the relative amount of reduction was found to be specific for each VDR agonist. Moreover, complex formation of the VDR on DNA resulted in a shift from the receptor's low-affinity ligand binding conformation (c3(LPD)) to its high affinity conformation (c1(LPD)). Finally, the characterization of the conformations of N- and C-terminally truncated VDR proteins defined the high-affinity ligand binding domain of the VDR as being positioned between amino acids 128 and 427. Taken together, the analysis of VDR conformations in solution in comparison to those of DNA-complexed VDR-RXR heterodimers allows a differentiation to be drawn between DNA-dependent and DNA-independent VD signaling pathways that can in turn be used for the identification of pathway selective VDR agonists.

Authors+Show Affiliations

Institut für Physiologische Chemie I and Biomedizinisches Forschungszentrum, Heinrich-Heine-Universität, D-40001 Düsseldorf, Germany.No affiliation info available

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

10648648

Citation

Quack, M, and C Carlberg. "The Impact of Functional Vitamin D(3) Receptor Conformations On DNA-dependent Vitamin D(3) Signaling." Molecular Pharmacology, vol. 57, no. 2, 2000, pp. 375-84.
Quack M, Carlberg C. The impact of functional vitamin D(3) receptor conformations on DNA-dependent vitamin D(3) signaling. Mol Pharmacol. 2000;57(2):375-84.
Quack, M., & Carlberg, C. (2000). The impact of functional vitamin D(3) receptor conformations on DNA-dependent vitamin D(3) signaling. Molecular Pharmacology, 57(2), pp. 375-84.
Quack M, Carlberg C. The Impact of Functional Vitamin D(3) Receptor Conformations On DNA-dependent Vitamin D(3) Signaling. Mol Pharmacol. 2000;57(2):375-84. PubMed PMID: 10648648.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - The impact of functional vitamin D(3) receptor conformations on DNA-dependent vitamin D(3) signaling. AU - Quack,M, AU - Carlberg,C, PY - 2000/1/29/pubmed PY - 2000/3/4/medline PY - 2000/1/29/entrez SP - 375 EP - 84 JF - Molecular pharmacology JO - Mol. Pharmacol. VL - 57 IS - 2 N2 - The vitamin D(3) receptor (VDR) is the nuclear receptor for 1alpha, 25-dihydroxyvitamin D(3) (VD) that acts primarily as a heterodimer with the retinoid X receptor (RXR) on different types of VD response elements, i.e., DNA-bound VDR-RXR heterodimers are the molecular switches in nuclear VD signaling pathways. In this study, DNA-dependent limited protease digestion assays and gel shift clipping assays were used for the analysis of VDR conformations and showed the same high ligand sensitivity for VD response element-bound VDR-RXR heterodimers (EC(50) of 0.1 nM for VD). In contrast, DNA-independent limited protease digestion assays clearly demonstrated a reduced ligand sensitivity for monomeric VDR in solution. Interestingly, the relative amount of reduction was found to be specific for each VDR agonist. Moreover, complex formation of the VDR on DNA resulted in a shift from the receptor's low-affinity ligand binding conformation (c3(LPD)) to its high affinity conformation (c1(LPD)). Finally, the characterization of the conformations of N- and C-terminally truncated VDR proteins defined the high-affinity ligand binding domain of the VDR as being positioned between amino acids 128 and 427. Taken together, the analysis of VDR conformations in solution in comparison to those of DNA-complexed VDR-RXR heterodimers allows a differentiation to be drawn between DNA-dependent and DNA-independent VD signaling pathways that can in turn be used for the identification of pathway selective VDR agonists. SN - 0026-895X UR - https://www.unboundmedicine.com/medline/citation/10648648/The_impact_of_functional_vitamin_D_3__receptor_conformations_on_DNA_dependent_vitamin_D_3__signaling_ L2 - http://molpharm.aspetjournals.org/cgi/pmidlookup?view=long&pmid=10648648 DB - PRIME DP - Unbound Medicine ER -