CAREER: Carbon-Detected NMR Methods for the Study of Intrinsically Disordered Proteins

职业：用于研究本质无序蛋白质的碳检测核磁共振方法

• 批准号: 0953918
• 负责人: Scott Showalter
• 金额: $ 96.84万
• 依托单位: Pennsylvania State Univ University Park
• 依托单位国家: 美国
• 项目类别: Continuing Grant
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-01-01 至 2015-12-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=0953918&HistoricalAwards=false
• 关键词: CAREER; Carbon; Detected; NMR; Methods

The objective of this CAREER project is to develop new models for protein conformational dynamics that encompass not only well-folded proteins, but lead to the application of structural biological tools to highly flexible macromolecular systems without defined structures, such as intrinsically disordered proteins (IDPs). A new NMR-based methodology will be implemented that utilizes carbon detected NMR to quantitatively characterize the properties of IDPs and the conformational transitions they often undergo upon binding other macromolecules. This research will yield new spectroscopic methods aimed at measuring conformational bias and dynamics in IDPs on the picoseconds-milliseconds timescales. The impact of the research protocol will be tested through studies of the microprocessor proteins DGCR8 and Drosha, which combine to affect the first step in the maturation of primary miRNA transcripts. Since its discovery ten years ago, miRNA has emerged as a major source of post-transcriptional gene regulation in multi-cellular organisms, but little is understood about the structural biology of the proteins involved in its biogenesis. The integrated education objective of this CAREER project is to utilize NMR spectroscopy as a powerful example for middle and high school students of basic biological and chemical principles. The PI's partnerships with Penn State's Take Our Daughters and Sons to Work Day program, and the Higher Achievement Program, which brings students from disadvantaged backgrounds to Penn State for a summer emersion experience, will continue. To enable off-site outreach for communities across Pennsylvania, a portable demonstration laboratory is being built around a Terranova-MRI Earth's Field NMR system. This system is the size of two car batteries, weighs less than 15 kg, and is controlled by a standard laptop computer. Utilizing the natural magnetic field of the earth for educational NMR spectroscopy allows the PI to bring an informal education program to nearby rural school districts or underserved urban centers including Harrisburg and Reading. There the PI will provide hands on demonstrations of magnetic resonance at the interface of biology and chemistry to middle and high school students. Demonstrations with the Terranova will be supported by demonstrations of fundamental concepts such as Faraday induction and the phenomenon of resonance. Demonstration materials used for these events will be made available to members of the Nittany Chemical Society for independent use in order to broaden the impact of these educational activities. In addition, the Terranova system will be used for in-class demonstrations at both the freshman chemistry and graduate biophysics levels in order to further strengthen the association between real world magnetic resonance and classroom content. The ultimate goal of the educational component is to pique student's interest in biological chemistry in an effort to increase enrollment of top students in college and graduate education in this area.

这个职业项目的目标是开发新的蛋白质构象动力学模型，不仅包括折叠良好的蛋白质，而且导致将结构生物学工具应用于高度灵活的没有定义结构的大分子系统，如内在无序蛋白质(IDPs)。将实施一种新的基于核磁共振的方法，利用碳检测到的核磁共振来定量表征IDPs的性质以及它们在与其他大分子结合时经常经历的构象转变。这项研究将产生新的光谱方法，旨在测量皮秒-毫秒时间尺度上国内流离失所者的构象偏差和动力学。研究方案的影响将通过对微处理器蛋白质Dgcr8和DROSHA的研究来测试，这两种蛋白质结合在一起会影响初级miRNA转录本成熟的第一步。自从十年前发现miRNA以来，它已经成为多细胞生物体转录后基因调控的主要来源，但对参与其生物发生的蛋白质的结构生物学知之甚少。这个职业项目的综合教育目标是利用核磁共振光谱学作为初中和高中学生基本生物和化学原理的有力范例。PI与宾夕法尼亚州立大学的带着我们的女儿和儿子去工作日计划以及更高成就计划的合作伙伴关系将继续下去，该计划将把来自弱势背景的学生带到宾夕法尼亚州立大学进行暑期再现体验。为了使宾夕法尼亚州的社区能够进行场外推广，围绕Terranova-MRI地球场核磁共振系统正在建造一个便携式演示实验室。该系统有两节汽车电池大小，重量不到15公斤，由一台标准的笔记本电脑控制。利用地球的自然磁场进行核磁共振波谱教育，使PI能够将非正式教育计划带到附近的农村学区或服务不足的城市中心，包括哈里斯堡和雷丁。在那里，PI将为初中生和高中生提供生物和化学界面上的磁共振演示。与Terranova的演示将得到诸如Faraday感应和共振现象等基本概念的演示的支持。将向Nittany化学学会成员提供用于这些活动的演示材料，供独立使用，以扩大这些教育活动的影响。此外，Terranova系统将用于新生化学和研究生生物物理学水平的课堂演示，以进一步加强现实世界磁共振与课堂内容之间的联系。教育部分的最终目标是激发学生对生物化学的兴趣，努力增加大学和该领域研究生教育的尖子生入学人数。