CAREER: Site-Directed Spin Labeling EPR Applications in Intrinsically Unstructured Proteins

该研究项目旨在利用基于电子顺磁共振(EPR)的方法来研究一类蛋白质的结构和构象，这些蛋白质被称为天然非结构化蛋白质，即它们没有稳定的三维结构。近三分之一的蛋白质被预测为非结构基序，很少有生物物理方法可以用来研究这类蛋白质。通过这项研究，将发展基于EPR的方法，并将研究两类蛋白质。第一种是一种基于多肽的天冬氨酸蛋白酶抑制剂。这些研究的结果对于了解天然多肽如何被用作酶抑制剂具有重要意义，从而可能为合理设计新的化学物质或基于多肽的抑制剂提供信息。第二种蛋白质参与调节许多细胞途径。这项研究的结果将为蛋白质如何在不采用明确的结构基序的情况下调节多个途径提供见解。本科生(特别是女性和那些来自代表性不足的群体)参与生物磁共振的研究，这项横跨生物、化学和物理领域的研究是这个将研究和教育目标联系起来的职业项目的中心主题。具体地说，这种集成是通过三层方法实现的。一方面是从本科生物化学和生物物理化学实验室招募学生进入研究型实验室，在那里学生可以直接将在课堂上学到的理论和实验技能应用到实际的研究环境中。第二个方面是针对大学一年级的本科生。在佛罗里达大学(UF)，已经有各种为此目的提供生源的计划：来自大学少数族裔指导计划(UMMP)的代表不足的少数族裔，来自UF本科生(Beckman Scholars，HHMI，REU)的少数族裔学生，以及来自UF大学学前教育和培训中心(CPET)组织的学生科学培训计划(SSTP)的高中生。第三层是与美国东南部的其他EPR小组开展暑期研究交流计划，旨在为本科生提供与EPR波谱相关的物理和生物学方面的跨学科培训。这个项目得到了化学部实验物理化学项目的共同资助。

The research project aims to utilize an electron paramagnetic resonance (EPR) based methodology to study the structure and conformation of a class of proteins that are termed natively unstructured, meaning they have no stable three dimensional structures. Nearly one third of proteins are predicted to function as unstructured motifs, and very few biophysical methods can be utilized to investigate this class of proteins. Through this research, the EPR based methods will be developed, and two classes of proteins will be investigated. The first is a peptide-based inhibitor of an aspartic protease. Results from these studies can be important for understanding how natural peptides are utilized as enzymatic inhibitors, thus potentially providing information about the rational design of new classes of chemicals or peptide-based inhibitors. The second protein is involved in regulating many cellular pathways. Results from this study will provide insights on how proteins can regulate multiple pathways without adopting a well defined structural motif.The participation of undergraduate students (particularly women and those from underrepresented groups) in research in biological magnetic resonance, which spans the fields of biology, chemistry and physics, serves as the central theme of this CAREER project that links research and educational objectives. Specifically, this integration is obtained by a three-tiered approach. One aspect is the recruiting of students into the research laboratory from undergraduate biochemistry and biophysical chemistry labs, where students can directly apply the theory and experimental skills learned in the classroom to a real research setting. The second aspect is the targeting undergraduate students within their first year of college. At The University of Florida (UF), a variety of programs are already in place that provide pools of students for this purpose: underrepresented minorities from the University Minorities Mentoring Program (UMMP), UF undergraduate students (Beckman Scholars, HHMI, REU), and high-school students from the Student Science Training Program (SSTP), organized by UF's Center for Pre-collegiate Education and Training (CPET). The third tier is the development of a summer research exchange program, with other EPR groups within the Southeastern USA, that aims to provide cross-discipline training for undergraduates in both physics and biology as related to EPR spectroscopy. This project is receiving co-funding from the Experimental Physical Chemistry Program of the Chemistry Division.