Screening for ligands of human retinoid X receptor-α.: Using ultrafiltration mass spectrometry
Screening for ligands of human retinoid X receptor-α.: Using ultrafiltration mass spectrometry
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DOI：
10.1021/ac701701k
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发表时间：
2007-12-15
影响因子：
7.4
通讯作者：
van Breemen, Richard B.
中科院分区：
文献类型：
--
作者：
Liu, Dongting;Guo, Jian;van Breemen, Richard B.
类视黄醇 X 受体 (RXR) 充当配体激活转录因子，是大量核受体异二聚体的必需成分。 RXR 有助于调节多种生理反应，包括细胞增殖、炎症、细胞分化和细胞凋亡的癌症预防反应。由于 RXR 代表癌症化学预防的重要靶点，因此开发了一种基于超滤质谱的测定法，以促进发现与人 RXR α 结合的潜在化学预防剂。 RXRa 的天然和合成配体（包括 9-顺式视黄酸、二十二碳六烯酸和 LG100268）可以在 DMSO（二甲基亚砜）甚至复杂基质（例如海洋细菌提取物）中检测和鉴定。通过使用超滤 LC-MS/MS（液相色谱-串联质谱法）的竞争性结合证明了配体与 RXR α 的特异性结合，并且可以按照对 RXR α 的亲和力顺序对配体进行排序。因此，超滤 LC-MS/MS 适用于筛选复杂混合物，例如天然产物提取物，以发现 RXR α 的新配体。
Retinoid X receptors (RXRs) function as ligand-activated transcription factors and are obligatory components of a large number of nuclear receptor heterodimers. RXRs help regulate diverse physiological responses including the cancer prevention responses of cell proliferation, inflammation, cell differentiation, and apoptosis. Since RXRs represent important targets for cancer chemoprevention, an ultrafiltration mass spectrometry-based assay was developed to facilitate the discovery of potential chemoprevention agents that bind to human RXR alpha. Natural and synthetic ligands for RXRa including 9-cis-retinoic acid, docosahexaenoic acid, and LG100268 could be detected and identified in DMSO (dimethyl sulfoxide) or even complex matrixes such as extracts of marine bacteria. Specific binding of ligands to RXR alpha was demonstrated through competitive binding using ultrafiltration LC-MS/ MS (liquid chromatography-tandem mass spectrometry), and ligands could be ranked in order of affinity for RXR alpha. Tberefore, ultrafiltration LC-MS/MS is suitable for the screening of complex mixtures such as natural product extracts for the discovery of new ligands to RXR alpha.