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

这个CAREER项目的目标是开发蛋白质构象动力学的新模型，不仅包括良好折叠的蛋白质，而且导致结构生物学工具应用于没有定义结构的高度灵活的大分子系统，如内在无序蛋白质（IDP）。将实施一种新的基于NMR的方法，利用碳检测NMR定量表征IDP的性质和它们在结合其他大分子时经常经历的构象转变。这项研究将产生新的光谱方法，旨在测量构象偏差和动态的皮秒-毫秒的时间尺度上的IDPs。研究方案的影响将通过对微处理器蛋白DGCR 8和Drosha的研究进行测试，DGCR 8和Drosha联合收割机影响初级miRNA转录物成熟的第一步。自10年前发现以来，miRNA已成为多细胞生物中转录后基因调控的主要来源，但对其生物发生所涉及的蛋白质的结构生物学知之甚少。这个CAREER项目的综合教育目标是利用NMR光谱学作为一个强有力的例子，为初中和高中学生的基本生物和化学原理。PI与宾夕法尼亚州立大学的“带我们的女儿和儿子去工作日”计划和“更高成就计划”的合作伙伴关系将继续下去，该计划将来自弱势背景的学生带到宾夕法尼亚州立大学进行夏季体验。为了使整个宾夕法尼亚州的社区能够进行场外推广，正在围绕Terranova-MRI地球场核磁共振系统建造一个便携式演示实验室。该系统的大小相当于两个汽车电池，重量不到15公斤，由一台标准笔记本电脑控制。利用地球的自然磁场进行教育核磁共振光谱学使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，包括哈里斯堡和阅读。在那里，PI将为初中和高中学生提供在生物和化学界面上的磁共振演示。演示与Terranova将支持演示的基本概念，如法拉第感应和共振现象。这些活动所用的演示材料将提供给尼塔尼化学学会的成员独立使用，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水平的课堂演示，以进一步加强真实的世界磁共振和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，以努力增加该领域大学和研究生教育的顶尖学生的入学率。

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.