CAA: Spectroscopic Analysis of the Tryptophan Residues of the BTK-SH3 Domain

SH3结构域是一个大约70个氨基酸的保守序列，通过与富含脯氨酸的多肽相互作用参与信号转导。这个职业进步奖(CAA)研究项目的目标是研究Bruton酪氨酸激酶SH3结构域的两个保守的色氨酸残基(BTK-SH3)在与富含脯氨酸的多肽相互作用中的具体作用。为了实现这一目标，将利用圆二色谱(CD)、荧光和拉曼光谱对BTK-SH3结构域野生型和单一氨基酸突变体进行表征，以阐明SH3结构域与富含Pro的多肽形成复合体时蛋白质结构的变化。拉曼光谱将确定由于BTK-SH3与富含Pro的多肽之间的相互作用而导致的色氨酸附近的微环境变化。这些研究将描述BTK-SH3结构域保守的色氨酸残基在分子水平上的作用及其在BTK-SH3结构域与调节富含Pro的多肽相互作用中的意义。这些研究的结果对于建立使用生物物理技术分析SH3领域的广泛和长期的研究项目将是至关重要的。广泛的影响：该项目将向代表性不足的学生打开实验室研究的大门，并帮助增加此类学生在本科水平的留存率。为了完成这项任务，将开展两项活动：面向少数民族大学生的暑期科学体验活动，以及面向4-6年级和7-9年级学生的为期一周的科学体验活动。为大学生提供的经验将允许他们在暑期和学年参与这个项目。学生将参加有关沟通的研究、研讨会、工作坊，并参观弗吉尼亚州的其他研究机构。学生将在地方、地区和国家科学会议上展示他们的研究成果。为期一周的暑期科学体验的参与者将享受适合他们年龄的课堂和实验室活动。学生将提高他们的数学和科学知识和技能。通过这些活动，学生将直接了解什么是在实验室工作，并将接触到科学的不同方面。这些活动的最终目标是吸引来自代表性不足的背景的学生获得生物和化学科学的本科学位。

The SH3 domain is a conserved sequence ~70 amino acids involved in signaling pathways through interactions with proline-rich peptides. The goal of this Career Advancement Award (CAA) research project is to investigate the specific roles of two conserved tryptophan residues of the Bruton's tyrosine kinase SH3 domain (BTK-SH3) in the interaction with proline rich peptides. To achieve this goal, BTK-SH3 domain wild type and single amino acid mutants will be characterized using circular dichroism (CD), fluorescence and Raman spectroscopies, which will elucidate protein structural changes upon complex formation between the SH3 domain and the proline-rich peptides. Raman spectroscopy will determine microenvironmental changes in the tryptophan vicinity due to the interaction between BTK-SH3 and the proline-rich peptides. The proposed studies will delineate the role of the conserved tryptophan residues of the BTK-SH3 domain at the molecular level and their significance in the interaction of the BTK-SH3 domain with regulating proline-rich peptides. The results from these studies will be critical in building a broad and long-term research project in analysis of SH3 domains using biophysical techniques.Broader impacts: This project will open the doors of laboratory research to underrepresented students and aid in increasing the retention of such students at the undergraduate level. To accomplish this task, two activities will be undertaken: a summer experience in science for college minority students and a weeklong science experience for 4th-6th and 7th-9th graders. The experience for the college students will allow them to participate in this project during the summer and academic year. Students will participate in research, seminars, workshops on communication and visits to other research facilities within Virginia. Students will present their research results in local, regional and national scientific meetings. The participants of the weeklong summer science experience will enjoy classroom and laboratory activities appropriate for their age. Students will enhance their mathematical and science knowledge and skills. Through these activities the students will know first hand what it is to work in a laboratory and will be exposed to different aspects of science. The end goal of these activities is to attract students from underrepresented backgrounds to obtain undergraduate degrees in biological and chemical sciences.