Minority Research Planning Grant - Spectroscopic Studies of the Interaction of the c-Src SH3 Domain With Proline-Rich Peptides

该项目的目标是确定c-Src蛋白的SH3结构域与富含Pro的多肽之间的相互作用所引起的结构变化。为了实现这些目标，将使用荧光共振能量转移(FRET)和傅里叶变换红外(FTIR)光谱。FRET实验将提供有关配体与SH3结构域疏水沟槽结合的距离的信息。SH3结构域结合槽包含一个保守的色氨酸残基，这将是拟议的实验的重点。FTIR实验将使我们能够研究蛋白质和多肽在形成络合物时的构象变化。这些研究的结合将有助于描绘SH3结构域和富含Pro的多肽之间的分子相互作用。更广泛的影响：拟议中的工作特别适合詹姆斯·麦迪逊大学的环境。P.I.在蛋白质和蛋白质表征方面拥有丰富的经验。化学系有许多本科专业的学生，他们对在学年和暑期进行研究非常感兴趣。此外，含有SH3结构域的蛋白质适合本科生使用。它们是可溶于水的蛋白质，从表达载体中纯化可产生大量蛋白质。本科生将学习蛋白质纯化和蛋白质工程技术。此外，学生还将获得光谱技术方面的经验，包括荧光和FTIR。接触不同的研究技术以及逻辑问题解决方面的培训将使学生获得新的科学职业道路。此外，一些建议的实验将成为一门普通生物化学实验室课程活动时间表的一部分。

The goal of this project is to identify structural changes induced by the interaction between the SH3 domain of the c-Src protein and proline-rich peptides. To achieve these goals, flourescence resonance energy transfer (FRET) and fourier transformed infrared (FTIR) spectroscopies will be used. FRET experiments will provide information about distances involved in ligand binding to the SH3 domain hydrophobic groove. The SH3 domain-binding groove contains a conserved tryptophan residue, which will be the focus of the experiments proposed. FTIR experiments will allow us to study conformational changes in both the protein and the peptide upon complex formation. The combination of these studies will allow delineation of molecular interactions between SH3 domains and proline-rich peptides. Broader Impact: The proposed work is particularly well suited for the environment at the James Madison University. The P.I. has extensive experience working with proteins and protein characterization. The Chemistry Department has a number of undergraduate majors who are very interested in pursuing research during the academic year as well as the summer months. Moreover, the SH3 domain-containing proteins are suitable for use by undergraduate students. They are water-soluble proteins and their purification from the expression plasmids yield large amounts of protein. The undergraduate students will learn protein purification and protein engineering techniques. Additionally, students will gain experience in spectroscopic techniques, including fluorescence and FTIR. The exposure to different research techniques as well as training in logical problem solving will give students access to new science career paths. Additionally, some of the proposed experiments will form part of the schedule of activities for a general biochemistry laboratory course.