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    • 47. 发明授权
    • Nuclear hormone receptor fluorescence polarization assay
    • 核激素受体荧光偏振测定
    • US06555326B1
    • 2003-04-29
    • US08975614
    • 1997-11-21
    • Kevin LustigPatrick BaeuerleHolger BeckmannJin-Long ChenBei Shan
    • Kevin LustigPatrick BaeuerleHolger BeckmannJin-Long ChenBei Shan
    • G01N3352
    • G01N33/542G01N33/74G01N2333/575G01N2333/70567
    • Methods for identifying modulators of nuclear hormone receptor function comprise the steps of (a) forming a mixture comprising a nuclear hormone receptor, a peptide sensor and a candidate agent, but not a natural coactivator protein of the receptor, wherein the sensor provides direct, in vitro binding to the receptor under assay conditions; (b) measuring an agent-biased binding of the sensor to the receptor; and (c) comparing the agent-biased binding with a corresponding unbiased binding of the sensor to the receptor. In particular embodiments, the sensor comprises an amphipathic alpha helix nuclear hormone interacting domain comprising a recited nuclear hormone transcriptional coactivator motif sequence, the sensor is present at sub-micromolar concentration, the binding reaction occurs in solution, the sensor comprises a fluorescent label and the measuring step comprises detecting fluorescence polarization of the label. Reagents include labeled sensor peptides and reaction mixtures consisting essentially of nuclear hormone receptor, a peptide and a candidate agent.
    • 用于鉴定核激素受体功能调节剂的方法包括以下步骤:(a)形成包含核激素受体,肽传感器和候选试剂但不是受体的天然共激活蛋白的混合物,其中所述传感器直接提供 在测定条件下与受体的体外结合; (b)测量传感器与受体的药剂偏置结合; 和(c)将代理偏好的结合与传感器与受体的相应的无偏见的结合进行比较。 在具体实施方案中,传感器包括包含所述核激素转录辅激动子序列的两亲性α螺旋核激素相互作用结构域,所述传感器以亚微摩尔浓度存在,所述结合反应发生在溶液中,所述传感器包含荧光标记和 测量步骤包括检测标签的荧光偏振。 试剂包括标记的传感器肽和基本上由核激素受体,肽和候选试剂组成的反应混合物。