Structural Studies of Proteins by Paramagnetic Solid-State NMR Spectroscopy

固态核磁共振（NMR）是一种光谱技术，其使得能够在大型复合物和组装体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NMR方法来确定蛋白质结构和相互作用，重点是测量长程结构约束，以克服与传统固态NMR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、本科生和高中生不同群体的教育和外联活动相结合。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计划，旨在通过向学生介绍化学，生物学和物理学接口的前沿跨学科研究，直接影响学生在教育经历的最早阶段。 该项目中固态NMR方法的特点在于其使用共价连接的顺磁性标签，这使得多个蛋白质位点能够同时获得高达~20 μ m的电子-核距离限制。具体而言，该项目旨在推进这种顺磁性固态NMR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进行蛋白质结构细化，探测大型生物大分子组装体中动态无序和结构化蛋白质结构域之间的重要功能相互作用，以及通过将顺磁固态NMR与低温动态核极化耦合来探索蛋白质中结构限制的测量，一个通用的工具，显着提高灵敏度的固态核磁共振谱。该项目由分子和细胞生物科学部的分子生物物理学小组支持。该奖项反映了NSF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。

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.