CRYSTALLOGRAPHIC STUDIES ON NUCLEAR PORE COMPLEX PROTEINS

这个子项目是许多研究子项目中利用 资源由NIH/NCRR资助的中心拨款提供。子项目和 调查员(PI)可能从NIH的另一个来源获得了主要资金， 并因此可以在其他清晰的条目中表示。列出的机构是 该中心不一定是调查人员的机构。 核孔复合体(NPC)是真核细胞中最大的蛋白质集合体，起着核质转运的调节作用。NPC大约有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个丙二醛蛋白质复合体。基于生化提纯和体外结合实验，所有核孔蛋白都被分成几个亚复合体。其中一个亚复合体Nup93亚复合体有助于NPC在核膜上的正确组装，并维持其大小排除限制。位于鼻咽癌核心区的Nup93亚复合体有Nup93、Nup205、Nup188、Nup35和Nup155五个成员，它们相互作用。我们的实验室正在尝试重建这个亚复合体，然后用结晶学方法确定它的三维结构。 我们实验室的长期目标是确定单个核孔蛋白或核孔素复合体的三维结构。这些蛋白质的结构信息将像是通过鼻咽癌进行核运输的快照。 目前，我们的Nup93和Nup35晶体的衍射率很低，我们的目标是收集良好的数据集，并使用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最大的蛋白质组装是如何组织的，以及调控核运输基本过程的蛋白质的结构特征是什么。最终，这些信息将引导我们在细胞水平上理解生命的复杂组织。

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. 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 methods The 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.