DETERMINATION OF THE PSEUDO-ATOMIC STRUCTURE OF NUCLEAR PORE COMPLEX (NPC) COMPO

核孔复合体（NPC）复合物拟原子结构的测定

• 批准号: 8362329
• 负责人: ANDREJ SALI
• 金额: $ 1.45万
• 依托单位: STANFORD UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-03-01 至 2012-02-29
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8362329
• 关键词: Architecture; Cell physiology; Complex; Computer software; Data; Funding; Genetic Transcription; Grant; Human; Malignant Neoplasms; Maps; Modeling; National Center for Research Resources; Nuclear Envelope; Nuclear Pore Complex; Nuclear Pore Complex Proteins; Play; Positioning Attribute; Principal Investigator; Proteins; Protocols documentation; Radiation; Relative (related person); Research; Research Infrastructure; Resolution; Resources; Sampling; Source; Structure; Tertiary Protein Structure; United States National Institutes of Health; Work; Yeasts; base; cost; human disease; improved; macromolecule; restraint; structural biology; three dimensional structure

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. Primary support for the subproject and the subproject's principal investigator may have been provided by other sources, including other NIH sources. The Total Cost listed for the subproject likely represents the estimated amount of Center infrastructure utilized by the subproject, not direct funding provided by the NCRR grant to the subproject or subproject staff. The Nuclear Pore Complex (NPC, about 50 MDa) is the sole passageway for the transport of macromolecules across the nuclear envelope. The pore plays a key role in numerous critical cellular processes such as transcription, and many of its components are implicated in human diseases such as cancer. Previous work provided the first description of the macromolecular architecture of the yeast NPC. This structure defined the relative positions and proximities of its 456 constituent nucleoporin (nup) proteins, based on spatial restraints derived from experimental data. Further elucidation of the evolutionary origin and transport mechanism of the NPC will require higher resolution information. To help improve upon the resolution and accuracy of the NPC structure, we propose to obtain small angle x-ray scattering (SAXS) data at SSRL. We are preparing a set of single protein, protein domain, and small NPC subcomplex samples for SAXS analysis. We will apply our Integrated Modeling Platform (IMP) software to incorporate a diverse set of experimental data, including SAXS spectra, as spatial restraints, to determine the three dimensional structures of these subcomplexes and proteins. We are specifically focusing on components of two subcomplexes, the 7-protein Nup84 subcomplex, and the 4-protein Nup170 complex, for which complementary experimental data are available. We have already used a similar protocol, to map different functional states of human Hsp90.

该子项目是利用资源的众多研究子项目之一 由 NIH/NCRR 资助的中心拨款提供。子项目的主要支持 并且子项目的主要研究者可能是由其他来源提供的， 包括其他 NIH 来源。 子项目可能列出的总成本 代表子项目使用的中心基础设施的估计数量， NCRR 赠款不直接向子项目或子项目工作人员提供资金。 核孔复合体（NPC，约 50 MDa）是大分子穿过核膜运输的唯一通道。孔在转录等许多关键细胞过程中发挥着关键作用，其许多成分与癌症等人类疾病有关。之前的工作首次描述了酵母 NPC 的大分子结构。该结构根据实验数据得出的空间限制，定义了其 456 种核孔蛋白 (nup) 蛋白的相对位置和接近度。进一步阐明 NPC 的进化起源和运输机制将需要更高分辨率的信息。为了帮助提高 NPC 结构的分辨率和准确性，我们建议在 SSRL 获取小角度 X 射线散射 (SAXS) 数据。我们正在准备一组用于 SAXS 分析的单一蛋白质、蛋白质结构域和小型 NPC 子复合体样品。我们将应用我们的集成建模平台 (IMP) 软件来整合一组不同的实验数据，包括 SAXS 光谱作为空间限制，以确定这些子复合物和蛋白质的三维结构。我们特别关注两个子复合物的组成部分，即 7 蛋白 Nup84 子复合物和 4 蛋白 Nup170 复合物，这两个子复合物有可用的补充实验数据。我们已经使用了类似的协议来绘制人类 Hsp90 的不同功能状态。