CRYSTALLOGRAPHIC STUDIES ON NUCLEAR PORE COMPLEX PROTEINS

核孔复合蛋白的晶体学研究

• 批准号: 8363347
• 负责人: GUNTER BLOBEL
• 金额: $ 0.31万
• 依托单位: BROOKHAVEN SCIENCE ASSOC-BROOKHAVEN LAB
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-07-01 至 2012-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8363347
• 关键词: Binding; Biochemical; Cell Nucleus; Complex; Crystallography; Cytoplasm; Data Set; Eukaryotic Cell; Exclusion; Funding; Goals; Grant; In Vitro; Individual; Life; Light; Mediating; Methods; Names; National Center for Research Resources; Nuclear; Nuclear Envelope; Nuclear Pore Complex; Nuclear Pore Complex Proteins; Principal Investigator; Process; Proteins; Research; Research Infrastructure; Resources; Source; Synchrotrons; United States National Institutes of Health; Vertebrates; base; cell assembly; cost; member; nucleocytoplasmic transport; protein complex; reconstitution; research study; three dimensional structure; trafficking

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) is the largest protein assembly in eukaryotic cell, which mediate transport between nucleus and cytoplasm. NPC have approximately 30 unique proteins (known as nucleoporins), clustered together to form approximately 125 MDa protein complex in vertebrates. Based on biochemical purification and in vitro binding experiments, all nucleoporins have been grouped into several subcomplexes. One of these subcomplex, named as Nup93 subcomplex contribute to correct assembly of NPC at nuclear membrane and also maintains its size exclusion limit. Nup93 subcomplex located in the core region of NPC has Nup93, Nup205, Nup188, Nup35 and Nup155 members, which interact with each other. Our lab is trying to reconstitute this subcomplex and then determine its 3D structure by crystallographic methods.The long term goal of our lab is to determine the 3D structure of individual nucleoporins or complex of nucleoporins.The structural information of these proteins will be like snapshots of nuclear trafficking via NPC.Currently we have crystals of Nup93 and Nup35 which are diffracting poorly.The goal is to collect good data set and determine its 3D structure using MAD/MIR methodsThe 3D structure of nucleoporins would demonstrate how the largest protein assembly of the cell is organized and what are structural features of the proteins which regulate the fundamental process of nuclear transport.Eventually this information will led us to understand the complex organization of life at cellular level.

这个子项目是许多利用资源的研究子项目之一 由NIH/NCRR资助的中心拨款提供。子项目的主要支持 子项目的主要研究者可能是由其他来源提供的， 包括其它NIH来源。 列出的子项目总成本可能 代表子项目使用的中心基础设施的估计数量， 而不是由NCRR赠款提供给子项目或子项目工作人员的直接资金。 核孔复合物（nuclear pore complex，NPC）是真核细胞中最大的蛋白质组装体，介导核质间的转运。NPC有大约30种独特的蛋白质（称为核孔蛋白），在脊椎动物中聚集在一起形成大约125 MDa的蛋白质复合物。基于生化纯化和体外结合实验，所有的核孔蛋白已被分成几个亚复合物。 Nup 93亚复合物是NPC在核膜上正确组装的亚复合物，并维持其分子排阻极限。 Nup 93亚复合体位于NPC的核心区域，由Nup 93、Nup 205、Nup 188、Nup 35和Nup 155 5个成员组成，它们之间相互作用。我们的实验室正在尝试重组这个亚复合体，然后通过晶体学方法确定其3D结构。我们实验室的长期目标是确定单个核孔蛋白或核孔蛋白复合体的3D结构。这些蛋白质的结构信息将像通过NPC的核运输的快照。目前我们有衍射较差的Nup 93和Nup 35晶体。目标是收集良好的数据集利用MAD/MIR方法确定其三维结构核孔蛋白的三维结构将展示细胞中最大的蛋白质组装体是如何组织的，以及调节核运输基本过程的蛋白质的结构特征，最终这些信息将引导我们从细胞水平理解生命的复杂组织。