Mapping nuclear pore surface with atomic force microscopy

用原子力显微镜绘制核孔表面

• 批准号: 16524566
• 负责人: Professor Dr. Hans Oberleithner
• 金额: --
• 依托单位: Institut für Physiologie II (Vegetative Physiologie)
• 依托单位国家: 德国
• 项目类别: Research Grants
• 财政年份: 2006
• 资助国家: 德国
• 起止时间: 2005-12-31 至 2008-12-31
• 项目状态: 已结题
• 来源: https://gepris.dfg.de/gepris/projekt/16524566?language=en
• 关键词: Mapping; nuclear; pore; surface; atomic

Nuclear pore complexes (NPCs) are supramolecular assemblies of proteins that span the nuclear envelope of eukaryotic cells. Transport of ions and macromolecules occurs through this pathway. It is still unclear whether different routes exist for different cargos and whether the access to such routes depends on the hydrophobic properties of the NPC surface. This open question is addressed by NPC surface mapping using atomic force microscopy (AFM). Surface scans across individual NPCs will be performed with narrow line spacing using AFM tips with different hydrophobic or hydrophilic properties. Derived from the AFM tip interaction with the NPC surface (i.e. adhesion forces) the hypothesis is tested whether the NPC surface consists of zones that separate transport routes into one for inorganic ions (hydrophilic route), one for small organic macromolecules (amphiphilic route) and one for large cargo using the central channel (hydrophobic route). A combination of electrical conductance and fluorescent dextran measurements will be used to selectively localize the individual pathways. With high resolution AFM imaging of the NPC surface we will search for the postulated peripheral channels in the NPC periphery. The goal of the project is a better understanding of nuclear translocation processes at the single NPC level.

核孔复合物（Nuclear Pore Complex，NPC）是真核细胞核膜上蛋白质的超分子组装体。离子和大分子的运输通过该途径发生。目前尚不清楚不同的货物是否存在不同的路线，以及这些路线的进入是否取决于NPC表面的疏水特性。这个悬而未决的问题是解决NPC表面映射使用原子力显微镜（AFM）。将使用具有不同疏水性或亲水性的AFM针尖以窄线间距对单个NPC进行表面扫描。从AFM针尖与NPC表面的相互作用（即粘附力）推导出的假设是测试NPC表面是否由区域组成，该区域将运输路线分为无机离子（亲水路线）、小有机大分子（两亲路线）和使用中央通道（疏水路线）的大货物。电导和荧光葡聚糖测量的组合将用于选择性地定位各个通路。用高分辨率AFM成像的NPC表面，我们将寻找假设的周边通道在NPC周边。该项目的目标是在单个NPC水平上更好地了解核易位过程。