STRUCTURAL INVESTIGATION OF HUMAN LIGAND BINDING DOMAIN BY SOLUTION NMR

这项工作的主要目的是研究构象 受体在与配体结合时的变化 抑制剂。雌激素受体是配体激活基因之一。 构成类固醇激素受体的调节蛋白 超级大家庭。这一蛋白质家族由三个不同的 结构域、N项、DNA结合结构域和配基结合结构域。在 没有激素(配体)，未连接的类固醇受体是 与热休克蛋白有关，这可能会促进激素 结合以及使DNA结合和/或反式激活失活 类固醇受体的功能。一个很大的构象变化是 已知激素受体在类固醇激素结合时。我们是 目前正在表达和纯化的配体结合域 雌激素受体。我们当前的目标是研究解决方案 用N15，C13多维核磁共振方法研究该蛋白质的结构 标记的、过氚的样品。在结构确定过程中 过程中，我们预计计算和图形的广泛使用 工具包括Sparky、Mardigras/Corma(完全松弛矩阵 分析)、MidasPlus和Chimera、Amber和X-Plor。

The main goal of this work is to investigate the conformational changes of the receptor upon binding of a ligand as well as an inhibitor. The estrogen receptor is one of the ligand-activated gene regulatory proteins which constitute the steroid-hormone receptor superfamily. This family of proteins are made of three distinct domains, N-term, DNA-binding, and ligand-binding domains. In the absence of hormone (ligand), the unliganded steroid receptor is associated with the heat shock protein, which may facilitate hormone binding as well as inactivate the DNA binding and/or transactivation functions of steroid receptors. A large conformational change is known to hormone receptors upon binding of steroid hormone. We are currently expressing and purifying the ligand-binding domain of the estrogen receptor. Our immediate goal is to study the solution structure of this protein by multi-dimensional NMR, using N15, C13 labeled, perdeuterated samples. During the structural determination process, we anticipate an extensive use of computational and graphics tools including Sparky, Mardigras/Corma (Complete Relaxation Matrix Analysis), MidasPlus and Chimera, Amber, and X-PLOR.