STRUCTURAL INVESTIGATION OF HUMAN LIGAND BINDING DOMAIN BY SOLUTION NMR

通过溶液核磁共振对人体配体结合域进行结构研究

• 批准号: 6119287
• 负责人: YOKO S HAGA
• 金额: $ 0.54万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
• 依托单位国家: 美国
• 财政年份: 1999
• 资助国家: 美国
• 起止时间: 1999-07-01 至 2000-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6119287
• 关键词: bacteria; biological products; biomedical resource; hormones; proteins

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.

这项工作的主要目标是研究构象 配体结合后受体的变化 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 超家族。 该蛋白质家族由三种不同的 域、N 端、DNA 结合域和配体结合域。 在 缺乏激素（配体），未配体的类固醇受体是 与热休克蛋白有关，可能促进激素 结合以及灭活 DNA 结合和/或反式激活 类固醇受体的功能。 较大的构象变化是 已知与类固醇激素结合后的激素受体。 我们是 目前表达并纯化了配体结合结构域 雌激素受体。 我们当前的目标是研究解决方案 使用 N15、C13​​ 通过多维 NMR 确定该蛋白质的结构 标记的、全氘化的样品。 在结构测定过程中 过程中，我们预计计算和图形的广泛使用 工具包括 Sparky、Mardigras/Corma（完整放松矩阵 Analysis）、MidasPlus 和 Chimera、Amber 和 X-PLOR。