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: 6280308
• 负责人: YOKO S HAGA
• 金额: $ 0.1万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
• 依托单位国家: 美国
• 财政年份: 1998
• 资助国家: 美国
• 起止时间: 1998-07-01 至 1999-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6280308
• 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.

本工作的主要目标是研究构象 受体在结合配体时的变化以及 抑制剂. 雌激素受体是一种配体激活的基因 构成类固醇激素受体的调节蛋白 超家族 这个蛋白质家族由三种不同的 结构域、N端、DNA结合和配体结合结构域。 在 激素（配体）的情况下，非配体类固醇受体是 与热休克蛋白有关，这可能有助于激素 结合以及抑制DNA结合和/或反式激活 类固醇受体的功能。 一个很大的构象变化是 已知激素受体结合类固醇激素。 我们 目前表达和纯化的配体结合结构域的 雌激素受体 我们的近期目标是研究解决方案 使用N15、C13通过多维NMR对该蛋白质进行结构分析 标记过的全氘样品 在结构测定过程中 在这个过程中，我们预计将广泛使用计算和图形 工具包括Sparky、Mardigras/Corma（完整的松弛矩阵 分析）、MidasPlus和Chimera、Amber和X-PLOR。