Mimicking the Nuclear Pore Complex with Protein Hydrogels

用蛋白质水凝胶模拟核孔复合体

• 批准号: 0906843
• 负责人: Lane Baker
• 金额: $ 40.69万
• 依托单位: Indiana University
• 依托单位国家: 美国
• 项目类别: Continuing Grant
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-09-01 至 2013-08-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=0906843&HistoricalAwards=false
• 关键词: Mimicking; Nuclear; Pore; Complex; Protein

ID: MPS/DMR/BMAT(7623) 0906843 PI: Baker, Lane ORG: Indiana UniversityTitle: Mimicking the Nuclear Pore Complex with Protein HydrogelsINTELLECTUAL MERIT: The nuclear pore complex (NPC) mediates transport of materials between the nucleus and cytoplasm of eukaryotic organisms. Current research indicates this unique transport mechanism relies on a protein-based hydrogel, which is both highly selective and capable of operating against concentration gradients. A well-resolved structural model of the NPC has been recently determined. This model and related reports have elucidated many aspects related to the mechanism of transport through the NPC, however critical features of this selective hydrogel phase are not understood. The long term goal of the proposal is to understand the fundamental mechanisms that govern the operation of this protein-hydrogel. The PI would also like to translate this knowledge to synthetic-based biomaterials capable of performing selective separations and sensing in ex-vivo platforms. The central hypothesis of this proposal is that by expressing selected proteins found in the NPC and rationally mutating the structure of these expressed proteins, a better picture of the molecular mechanisms of transport through the NPC can be developed. Further, it is hypothesized that this enhanced understanding will allow development of model semi-synthetic polypeptide hydrogels that will display biomimetic transport properties. This hypothesis has been formulated based on literature reports and preliminary studies, which show proteins from the NPC can be coerced to form selective hydrogels in ex-vivo environments. Specifically, the PI will: (1) determine the relative interactions and properties of proteins located within the nuclear pore complex that are necessary to reproduce selective transport observed in the nuclear pore complex, (2) determine the influence of the hydrophilic regions of the protein hydrogel on transport and the extent to which these regions can be used to synthetically alter transport properties, and (3) develop a route for generating a semi-synthetic hydrogel using solid phase peptide synthesis. BROADER IMPACTS: Biological systems offer remarkable insight and inspiration for rational materials design. To advance current capabilities, fundamental advances in how biological mechanisms, such as nucleocytoplasmic transport, operate must be realized. The themes upon which this research is based ? biomaterials and bio/analytical chemistry ? can be used to engage students, especially if the subject matter is approached in an appropriate manner. To this end, the PI has initiated an educational outreach program that is integrated with the subject matter of the research program and involves the participation of graduate students in the group. The outreach program uses visual learning concepts and information dissemination using video conferencing in coordination with podcasting technology, forming specific modules that consist of a podcast, a classroom exercise, a follow-up live video conference, and a short assessment of the exercise. Initially, they are focusing on collaboration with two high schools, one of which serves predominantly underrepresented groups. Both high schools are located hundreds of miles (1300 and 800, respectively) from the PI?s laboratory. This is intended to prove that the educational program under development can use the internet effectively to bridge both geographic and demographic divides. To increase the impact of the program the PI makes initial personal visits to the collaborating schools and participates in interactive question-and answer sessions throughout the semester using video conferencing. Along with this collaboration, additional content will be developed and disseminated in separate podcasts that specifically target chemistry students at the high school, undergraduate, and graduate levels. Work is also disseminated through peer-reviewed journals. Pedagogical assessment of the program will be performed in collaboration with Indiana University campus facilities.

ID：MPS/DMR/BMAT（7623）0906843 PI：BAKER，LANE ORG：INDIANA UniversityTitle：用蛋白质的水凝胶功能（NPC）介导核和细胞质质体细胞质体之间的材料传输。当前的研究表明，这种独特的运输机制依赖于基于蛋白质的水凝胶，该水凝胶既具有高度选择性，又能够在浓度梯度上进行操作。最近已经确定了NPC的完善结构模型。该模型和相关的报告阐明了与通过NPC传输机制相关的许多方面，但是该选择性水凝胶相的关键特征尚不清楚。该提案的长期目标是了解控制这种蛋白质凝胶凝胶运行的基本机制。 PI还希望将此知识转化为基于合成的生物材料，能够在前体内平台中进行选择性分离和感测。该提议的中心假设是，通过表达在NPC中发现的选定蛋白质并合理地突变这些表达蛋白的结构，可以更好地了解通过NPC传输的分子机制。此外，假设这种增强的理解将允许开发模型的半合成多肽水凝胶，这些水凝胶将显示出仿生运输特性。该假设是根据文献报告和初步研究提出的，该研究表明，可以强制将NPC的蛋白质胁迫在前体内环境中形成选择性水凝胶。 具体而言，PI将：（1）确定位于核孔复合体中蛋白质的相对相互作用和特性，这些蛋白质是在核孔复合物中观察到的选择性转运所必需的，（2）确定蛋白质水凝胶的亲水区域的影响，以及这些区域对这些区域的使用程度，可用于使用固体构图的综合性（3），并在核孔复合物中产生（3）。肽合成。更广泛的影响：生物系统为理性材料设计提供了非凡的见识和灵感。为了提高当前功能，必须实现生物学机制（例如核质运输）的基本进步。这项研究基于的主题？生物材料和生物/分析化学？可以用来吸引学生，特别是如果以适当的方式接触主题。为此，PI启动了与研究计划的主题相结合的教育外展计划，并涉及研究生参与小组。该外展计划使用视频会议使用视觉学习概念和信息传播，以与播客技术协调，形成由播客，课堂练习，后续视频会议以及对练习的简短评估的特定模块。最初，他们专注于与两所高中的合作，其中一所主要服务于代表性不足的群体。两所高中分别位于PI的实验室数百英里（分别为1300和800英里）。这旨在证明，正在开发的教育计划可以有效地利用互联网来弥合地理和人口鸿沟。为了增加该计划的影响，PI首次对合作学校进行了个人访问，并在整个学期中使用视频会议参加了交互式问答课程。除此合作外，还将在单独的播客中开发和传播其他内容，这些播客专门针对高中，本科和研究生级别的化学学生。工作也通过同行评审的期刊传播。该计划的教学评估将与印第安纳大学校园设施合作进行。