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）的方法来研究一类被称为天然非结构化的蛋白质的结构和构象，这意味着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。通过本研究，将开发基于EPR的方法，并对两类蛋白质进行研究。第一种是基于肽的天冬氨酸蛋白酶抑制剂。这些研究的结果对于理解天然肽如何被用作酶抑制剂很重要，因此可能为合理设计新型化学品或肽基抑制剂提供信息。第二种蛋白质参与调节许多细胞通路。这项研究的结果将为蛋白质如何在不采用明确定义的结构基序的情况下调节多种途径提供见解。本科学生（特别是女性和那些来自代表性不足群体的学生）参与生物磁共振研究，这跨越了生物学、化学和物理学领域，是这个职业项目的中心主题，它将研究和教育目标联系起来。具体来说，这种集成是通过三层方法实现的。一个方面是招收学生从本科生物化学和生物物理化学实验室进入研究实验室，学生可以直接将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以内的本科生。在佛罗里达大学（UF），各种各样的项目已经为这一目的提供了学生库：来自大学少数民族指导计划（UMMP）的未被充分代表的少数民族，佛罗里达大学本科生（贝克曼学者，HHMI， REU），以及由佛罗里达大学大学预科教育和培训中心（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小组开展夏季研究交流计划，旨在为物理学和生物学的本科生提供与EPR光谱相关的跨学科培训。本项目由化学学部实验物理化学项目共同资助。

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.