Structural Studies of Proteins by Paramagnetic Solid-State NMR Spectroscopy

通过顺磁固态核磁共振波谱法研究蛋白质的结构

• 批准号: 2303862
• 负责人: Christopher Jaroniec
• 金额: $ 93.21万
• 依托单位: Ohio State University
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-07-01 至 2027-06-30
• 项目状态: 未结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=2303862&HistoricalAwards=false
• 关键词: Structural; Studies; Proteins; Paramagnetic; Solid

Solid-state nuclear magnetic resonance (NMR) is a spectroscopic technique, which enables atomistic studies of structure, dynamics, and interactions to be performed for proteins and nucleic acids in the context of large complexes and assemblies. Such systems, which include membrane-associated proteins, oligomeric molecular machines, amyloids, and chromatin, play important roles in fundamental biological processes and mechanisms. In this project, the PI is developing and applying new solid-state NMR methods for the determination of protein structures and interactions, with focus on measurements of long-range structural restraints to overcome some of the major challenges associated with conventional solid-state NMR approaches. The research aims of the project are integrated with education and outreach activities involving diverse groups of graduate, undergraduate and high-school students. These activities, which include intensive, full-time summer research internship programs for undergraduate and local high-school students organized annually in the PI's laboratory, aim to directly impact students in the earliest stages of their educational experience by introducing them to cutting edge interdisciplinary research at the interface of chemistry, biology, and physics. The feature of the solid-state NMR methodology in this project lies in its use of covalently attached paramagnetic tags, which enable electron-nucleus distance restraints up to ~20 Å to be simultaneously accessed for multiple protein sites. Specifically, the project aims to advance this paramagnetic solid-state NMR methodology for protein structure refinement in systems of increasing complexity including amyloid fibrils and oligomeric protein complexes, probing functionally-important interactions between dynamically disordered and structured protein domains in large biomacromolecular assemblies, and exploring measurements of structural restraints in proteins by coupling paramagnetic solid-state NMR with low-temperature dynamic nuclear polarization, a general tool for dramatically enhancing the sensitivity of solid-state NMR spectra. This project is supported by the Molecular Biophysics Cluster of the Molecular and Cellular Biosciences Division.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.

固态核磁共振（NMR）是一种光谱技术，它可以在大型复合物和组装的背景下对蛋白质和核酸进行结构、动力学和相互作用的原子研究。这些系统包括膜相关蛋白、寡聚分子机器、淀粉样蛋白和染色质，在基本的生物过程和机制中发挥重要作用。在这个项目中，PI正在开发和应用新的固态核磁共振方法来确定蛋白质结构和相互作用，重点是远程结构约束的测量，以克服传统固态核磁共振方法相关的一些主要挑战。该项目的研究目标与涉及研究生、本科生和高中生的不同群体的教育和外展活动相结合。这些活动，包括每年在PI实验室为本科生和当地高中生组织的密集的全日制暑期研究实习项目，旨在通过向学生介绍化学，生物和物理界面的前沿跨学科研究，直接影响他们教育经历的早期阶段。固态核磁共振方法在这个项目中的特点在于它使用共价连接的顺磁标签，这使得电子-核距离限制高达~20 Å可以同时访问多个蛋白质位点。具体来说，该项目旨在推进这种顺磁固态核磁共振方法，用于在包括淀粉样蛋白原纤维和寡聚蛋白复合物在内的日益复杂的系统中改进蛋白质结构，探测大型生物大分子组装中动态无序和结构化蛋白质结构域之间的功能重要相互作用。以及通过将顺磁固态核磁共振与低温动态核极化（一种显著提高固态核磁共振光谱灵敏度的通用工具）耦合来探索蛋白质结构约束的测量。本项目由分子与细胞生物科学部分子生物物理组支持。该奖项反映了美国国家科学基金会的法定使命，并通过使用基金会的知识价值和更广泛的影响审查标准进行评估，被认为值得支持。