Boehm M F, McClurg M R, Pathirana C, Mangelsdorf D, White S K, Hebert J, Winn D, Goldman M E, Heyman R A
Department of Medicinal Chemistry, Ligand Pharmaceuticals, Incorporated, San Diego, California 92121.
J Med Chem. 1994 Feb 4;37(3):408-14. doi: 10.1021/jm00029a013.
all-trans-Retinoic acid is known to bind to the retinoic acid receptors (RARs) resulting in an increase in their transcriptional activity. In contrast, recently identified 9-cis-retinoic acid (9-cis-RA), which is an additional endogenous RA isomer, is capable of binding to both RARs and retinoid X receptors (RXRs). These distinct properties have raised questions as to the biological role governed by these two retinoic acid isomers and the set of target genes that they regulate. Herein, we report the synthesis of high specific activity [3H]-9-cis-RA and its application to study the ligand-binding properties of the various retinoid receptor subtypes. We examined the binding properties of RARs and RXRs for a series of synthetic retinoids and compared the ligand-binding properties of these arotinoid analogs with their ability to regulate gene expression via the retinoid receptors in a cotransfection assay. The utilization of the [3H]-9-cis-RA competitive binding assay and the cotransfection assay has made it possible to rapidly identify important structural features of retinoids leading to increased selectivity for either the RAR or RXR receptor subtypes.
众所周知，全反式维甲酸可与维甲酸受体（RARs）结合，从而增加其转录活性。相比之下，最近发现的9-顺式维甲酸（9-cis-RA）是另一种内源性维甲酸异构体，它既能与RARs结合，也能与维甲酸X受体（RXRs）结合。这些不同的特性引发了关于这两种维甲酸异构体所调控的生物学作用以及它们所调节的靶基因集的问题。在此，我们报告了高比活度[3H]-9-顺式维甲酸的合成及其在研究各种类维生素A受体亚型配体结合特性中的应用。我们研究了RARs和RXRs对一系列合成类维生素A的结合特性，并在共转染实验中比较了这些类维生素A类似物的配体结合特性与其通过类维生素A受体调节基因表达的能力。[3H]-9-顺式维甲酸竞争性结合实验和共转染实验的应用使得快速鉴定导致对RAR或RXR受体亚型选择性增加的类维生素A重要结构特征成为可能。
