Structural Studies of Proteins by Paramagnetic Solid-State NMR Spectroscopy

点击翻译按钮获取中文摘要

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 bio-molecular (supramolecular) assemblies. A wide range of such assemblies, which include amyloids (molecular assembiles responsible for neurodegenerative diseases), lipid membrane protein complexes and chromatin, play important roles in fundamental biological processes and mechanisms. In this project, the PI is developing new solid-state NMR methodologies for the determination of protein structures and interactions, which focus on measurements of long-range structural restraints and overcome some of the key challenges associated with conventional solid-state NMR techniques. 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, 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.The unique feature of the proposed novel NMR methods being developed lies in their use of covalently-attached paramagnetic tags, which enable up to ~20 Å electron-nucleus distance restraints to be simultaneously accessed for multiple protein sites via multidimensional NMR approaches. Already having recently demonstrated that it is possible to elucidate protein structures in a de novo manner by using paramagnetic solid-state NMR, the current project aims to further advance this methodology by developing compact and versatile metal binding tags that permit measurements of a broad range of paramagnetic effects, systematically exploring the limits of high-resolution protein structure determination by paramagnetic solid-state NMR, and expanding the realm of applicability of this technology beyond model globular proteins in microcrystalline phase to large assemblies of biological macromolecules.