CAREER: Structure-Specific Fluorescence Spectroscopy to Dissect Conformational Heterogeneity in Macromolecules

职业：结构特异性荧光光谱分析大分子的构象异质性

• 批准号: 2338251
• 负责人: Julia Widom
• 金额: $ 66.2万
• 依托单位: University of Oregon Eugene
• 依托单位国家: 美国
• 项目类别: Continuing Grant
• 财政年份: 2024
• 资助国家: 美国
• 起止时间: 2024-06-01 至 2029-05-31
• 项目状态: 未结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=2338251&HistoricalAwards=false
• 关键词: CAREER; Structure; Specific; Fluorescence; Spectroscopy

With support from the Chemical Measurement and Imaging Program in the Division of Chemistry, and partial co-funding from the Molecular Biophysics Cluster in the Division of Molecular and Cellular Biosciences, Dr. Julia Widom and her research group at the University of Oregon (UO) are developing measurement and analysis methods to resolve with high spatial and temporal resolution the diverse structures adopted by biological macromolecules. Biological macromolecules frequently transition across multiple conformational structures, and the nature and prevalence of these structures is of great significance to both their intrinsic biological functions, and their potential applications as drug targets, biomarkers, and building blocks in nanostructures. With a focus on RNA and DNA, the Widom Lab is establishing methods based on high-resolution laser spectroscopy that extract distinct signatures from macromolecules with different structures, and is expanding the toolbox of well-characterized molecular probes that can be used for such measurements. In addition, Dr. Widom is partnering with the UO undergraduate Chemistry Club to engage in outreach to middle- and high-school students in rural Oregon communities through remote, interactive chemistry activities, site visits to UO by rural students, and collaboration with the chemistry program at Eastern Oregon University.Most existing spectroscopic techniques are sensitive to either the local (Angstrom length-scale) or global (nanometer length-scale) structure of the system being measured. To overcome this limitation, Dr. Widom’s research group is developing a method based on measurement of fluorescence spectra on an ultrafast timescale, utilizing excited-state energy transfer between two probes to separate signals from tightly and loosely folded conformational subpopulations. Global fitting of the resulting time-resolved emission spectra connects the local environments of the two probes to the global conformations of the macromolecule within which they reside. Structured DNA and RNA oligomers with at least two well-defined global conformations will be used as model systems, followed by expansion of the technique to study the binding of small molecules to RNA. The Widom Lab will also investigate the suitability of different fluorescent probes for applications involving nucleic acids, analyzing their photophysical properties and their impacts on RNA structure. If successful, this project has the potential to provide new tools for the analysis of structural heterogeneity in complex chemical, biological and material systems and thus have far-reaching scientific broader impact.This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.

在化学系化学测量和成像项目的支持下，以及分子和细胞生物科学系分子生物物理学集群的部分共同资助下，俄勒冈大学 (UO) Julia Widom 博士和她的研究小组正在开发测量和分析方法，以高空间和时间分辨率解析生物大分子采用的多种结构。生物大分子经常跨越多种构象结构，并且这些结构的性质和普遍性非常重要。 Widom 实验室以 RNA 和 DNA 为重点，正在建立基于高分辨率激光光谱的方法，以提取不同的特征。 Widom 博士正在与俄勒冈大学本科生化学俱乐部合作，向中学生和高中生进行推广。通过远程互动化学活动、农村学生对俄勒冈大学的实地考察以及与东俄勒冈大学化学项目的合作，在俄勒冈州农村社区进行了光谱技术的研究。大多数现有的光谱技术对局部（埃长度尺度）或全球（纳米）敏感为了克服这一限制，Widom 博士的研究小组正在开发一种基于超快时间尺度荧光光谱测量的方法，利用两个探针之间的激发态能量转移来紧密分离信号。和松松地折叠所得到的时间分辨发射光谱的全局拟合将两个探针的局部环境与它们所在的大分子的全局构象连接起来，将使用具有至少两个明确的全局构象的结构化DNA和RNA寡聚物。作为模型系统，随后扩展研究小分子与 RNA 结合的技术，Widom 实验室还将研究不同荧光探针在涉及核酸的应用中的适用性，分析其光物理特性及其对 RNA 的影响。如果成功，该项目有可能为复杂化学、生物和材料系统中的结构异质性分析提供新的工具，从而产生深远的科学影响。该奖项是 NSF 的法定使命，并被认为值得支持。通过使用基金会的智力优点和更广泛的影响审查标准进行评估。