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

CAA：BTK-SH3 结构域色氨酸残基的光谱分析

• 批准号: 0542288
• 负责人: Rosa Rivera-Hainaj
• 金额: $ 17.48万
• 依托单位: James Madison University
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-03-01 至 2006-12-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=0542288&HistoricalAwards=false
• 关键词: CAA; Spectroscopic; Analysis; Tryptophan; Residues

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.

SH3 结构域是约 70 个氨基酸的保守序列，通过与富含脯氨酸的肽相互作用参与信号传导途径。该职业进步奖 (CAA) 研究项目的目标是研究布鲁顿酪氨酸激酶 SH3 结构域 (BTK-SH3) 的两个保守色氨酸残基在与富含脯氨酸的肽相互作用中的具体作用。为了实现这一目标，BTK-SH3结构域野生型和单氨基酸突变体将使用圆二色性（CD）、荧光和拉曼光谱进行表征，这将阐明SH3结构域和富含脯氨酸的肽之间形成复合物时蛋白质结构的变化。拉曼光谱将确定由于 BTK-SH3 和富含脯氨酸的肽之间的相互作用而导致色氨酸附近的微环境变化。拟议的研究将在分子水平上描述 BTK-SH3 结构域的保守色氨酸残基的作用及其在 BTK-SH3 结构域与调节富含脯氨酸的肽相互作用中的重要性。这些研究的结果对于建立一个利用生物物理技术分析 SH3 领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，并有助于提高此类学生在本科阶段的保留率。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，为四年级至六年级和七至九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与该项目。学生将参加研究、研讨会、交流讲习班以及参观弗吉尼亚州的其他研究设施。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。为期一周的夏季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，学生将直接了解在实验室工作是什么，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弱势背景的学生获得生物和化学科学的本科学位。