Structural studies of proteins by paramagnetic solid-state NMR spectroscopy

在这个项目中，PI正在开发新的多维核磁共振波谱技术，用于确定固相中的三维蛋白质结构。这些核磁共振方法依赖于共价连接的顺磁标签的使用，并利用大的电子-核超精细相互作用来同时在多个蛋白质位置获得长达~20？的电子-核距离限制。该项目的主要研究目标是：开发以增强灵敏度的基于顺磁的核磁共振结构限制的收集方法，开发用于探测大型蛋白质组件的高阶分子间结构的方法，以及设计紧凑的顺磁标签以提高使用该方法得出的蛋白质结构的质量。这个项目产生的顺磁固体核磁共振技术有望直接应用于各种大分子组件的结构分析，这些组件是基本生物过程的核心，但不容易使用传统的高分辨率结构工具进行表征。该项目的研究目标是与教育和推广活动相结合。小学教育活动将包括每年在PI的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。这些项目的招生将特别关注STEM学科中传统上代表性不足的群体的学生。这些活动的主要目的是通过向学生介绍化学、生物和物理相结合的尖端跨学科研究，建立他们对自己和他们的科学能力的信心，并鼓励他们以一种广泛的、基于发现的方式来思考科学，从而在他们教育经历的早期阶段直接影响他们。最终目标是增加学生在STEM领域的参与度和留存率。该项目由分子和细胞生物科学部的分子生物物理组和化学部的生命过程化学项目共同支持。

In this project the PI is developing new multidimensional NMR spectroscopy techniques for the determination of three-dimensional protein structures in the solid phase. These NMR methods rely on the use of covalently-attached paramagnetic tags and exploit the large electron-nucleus hyperfine interactions to simultaneously obtain long, up to ~20 Å, electron-nucleus distance restraints at multiple protein sites. The principal research objectives of the project are: to develop methodology for the collection of paramagnetic-based NMR structural restraints with enhanced sensitivity, to develop methodology for probing the higher order intermolecular architecture of large protein assemblies, and to design compact paramagnetic tags to improve the quality of protein structures derived using this methodology. The paramagnetic solid-state NMR techniques emerging from this project are expected to be directly applicable to the structural analysis of a variety of large macromolecular assemblies that are central to fundamental biological processes yet do not readily lend themselves to characterization using traditional high-resolution structural tools.The research aims of the project are integrated with education and outreach activities. The primary educational activity will consist of intensive, full-time summer research internship programs for undergraduate and local high-school students organized annually in the PI's laboratory. Student recruitment for these programs will focus particularly on students from groups traditionally under-represented in the STEM disciplines. The main objective of these activities is 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, building their confidence in themselves and their scientific abilities, and encouraging them to think of science in a broad, discovery-based manner. The ultimate goal is to increase student participation and retention in the STEM fields. This project is jointly supported by the Molecular Biophysics Cluster in the Division of Molecular and Cellular Biosciences and by the Chemistry of Life Processes Program in the Chemistry Division.