EAPSI: Understanding How a Complex Cellular Environment Influences Amyloid Fibril Protein Dynamics

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This project supports research in a collaboration with Prof. Xueqian Kong, an expert on studying materials using the cutting-edge technique of fast magic angle spinning-nuclear magnetic resonance (MAS-NMR), in the department of Chemistry, Zhejiang University in Hangzhou to develop innovative methodology in the field of biological solid-state NMR. These new methods could significantly enhance the spectral sensitivity in biological NMR samples. Over the 8-week funding period, we will focus on studying protein dynamics in complex cellular environments. Current solid-state NMR techniques suffers from the poor sensitivity because it reads out information on low gamma nuclei (e.g. Carbon-13). This can be solved by using a sensitivity enhancing technique, dynamic nuclear polarization (DNP) NMR, but this must be done at extremely cold temperatures (100 K), which eliminate dynamic motions. The current collaborative project is to learn and apply fast MAS-NMR which reports on higher gamma nuclei (e.g. Hydrogen-1), performed at room temperature, to study the protein dynamics of amyloid fibrils in their native cellular environments. The broader impact of this project includes applying these methods to study fibrils formed by proteins such as tau, which is implicated in Alzheimer's disease.This project will encompass a balanced approach that uses the innovative sample preparation methods developed in the home lab and sound methodology and instrumentation to demonstrate significant sensitivity enhancement in biomolecular NMR during the EAPSI research at Zhejiang University. This work will result two major findings i) if fast MAS-NMR can be used to study proteins at endogenous levels and ii) determine if the cellular environment affect protein dynamics, particularly of intrinsically disordered regions. The intellectual merits of the proposed research lie in establishing fast MAS-NMR as a complementary technique to DNP-NMR for studying proteins at endogenous levels in complex cellular environments. Its broader impacts lie in developing alternative methods that will allow for the further study of intrinsically disordered proteins that make fibrils that are often implicated in disease states.This award, under the East Asia and Pacific Summer Institutes program, supports summer research by a U.S. graduate student and is jointly funded by NSF and the Chinese Ministry of Science and Technology.