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.
