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

SH 3结构域是一个保守序列，约70个氨基酸，通过与富含脯氨酸的肽相互作用参与信号传导途径。这个职业发展奖（CAA）的研究项目的目标是调查两个保守的色氨酸残基的布鲁顿的酪氨酸激酶SH 3结构域（BTK-SH 3）在与脯氨酸丰富的肽相互作用的具体作用。为了实现这一目标，将使用圆二色性（CD）、荧光和拉曼光谱来表征BTK-SH 3结构域野生型和单个氨基酸突变体，这将阐明SH 3结构域和富含脯氨酸的肽之间形成复合物后的蛋白质结构变化。拉曼光谱将确定由于BTK-SH 3和富含脯氨酸的肽之间的相互作用而引起的色氨酸附近的微环境变化。这些研究将在分子水平上阐明BTK-SH 3结构域保守色氨酸残基的作用及其在BTK-SH 3结构域与富含脯氨酸的调节肽相互作用中的意义。这些研究的结果对于建立一个使用生物物理技术分析SH 3结构域的广泛而长期的研究项目至关重要。更广泛的影响：该项目将为代表性不足的学生打开实验室研究的大门，并有助于提高这些学生的保留率本科水平。为了完成这项任务，将开展两项活动：为少数民族大学生提供暑期科学体验，为4 - 6年级和7 - 9年级学生提供为期一周的科学体验。大学生的经验将使他们能够在夏季和学年期间参加这个项目。学生将参加研究，研讨会，交流和参观弗吉尼亚州内的其他研究设施讲习班。学生将在地方，区域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，学生将知道第一手资料是什么，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.