Identification of a competitive binding component in vitamin D-resistant New World primate cells with a low affinity but high capacity for 1,25-dihydroxyvitamin D3.
Monkeys in a number of different New World primate genera express a form of compensated target organ resistance to steroid hormones, including 1,25-dihydroxyvitamin D3 [1,25-(OH)2D3]. Characterization of these phenotypes has previously relied upon the study of the 1,25-(OH)2D3-receptor (VDR) interaction in cultured dermal fibroblasts from affected primates. In this report, we show that three of these prototypic phenotypes can be faithfully reproduced in previously established cultured cell lines: B95-8, EBV-transformed B lymphoblasts from the marmoset (Callithrix jacchus), a New World primate with recognized vitamin D resistance; OMK, renal tubular epithelial cells from the owl monkey (Aotus trivergatus), a New World primate with an Old World primate-like VDR phenotype; and MLA144, transformed B lymphoblasts from a gibbon (Hylobates), an Old World primate that expresses the wild-type VDR phenotype. The rank order of specific nuclear uptake and binding of [3H]1,25-(OH)2D3 to the VDR was OMK > or = MLA144 >> B95-8. Despite a 7- to 9-fold difference in cellular VDR content according to ligand binding analyses, there was no discernible difference in the internalization constant Kin for specific cellular uptake of [3H]1,25-(OH)2D3 (0.12-0.26 nM) or in the quantity of VDR detected by immunoblot analysis. We now speculate that the discrepancy in VDR quantitation by binding and immunoblot analysis in the B95-8 New World primate cell line results from the presence of an intracellular, vitamin D metabolite binding moiety in this cell line that competes with the VDR for metabolite binding.(ABSTRACT TRUNCATED AT 250 WORDS)
Division of Endocrinology, Cedars-Sinai Medical Center, University of California-Los Angeles School of Medicine.
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Adams
D000818:Animals; D016645:Aotidae; D001667:Binding, Competitive; D002117:Calcitriol / Q000378:metabolism* / Q000494:pharmacology; D002144:Callithrix; D002460:Cell Line; D002461:Cell Line, Transformed; D002851:Chromatography, High Pressure Liquid; D002852:Chromatography, Ion Exchange; D004351:Drug Resistance; D005347:Fibroblasts / Q000187:drug effects*; D006922:Hylobates; D010641:Phenotype; D018167:Receptors, Calcitriol; D011987:Receptors, Steroid / Q000378:metabolism*
D018167:Receptors, Calcitriol; D011987:Receptors, Steroid; D002117:Calcitriol