Structural Studies of Proteins by Paramagnetic Solid-State NMR Spectroscopy

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.