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

该项目支持与杭州浙江大学化学系利用快速魔角旋转-核磁共振（MAS-NMR）尖端技术研究材料的专家孔学谦教授合作进行研究，以开发生物固态NMR领域的创新方法。这些新方法可以显著提高生物NMR样品的光谱灵敏度。在为期8周的资助期间，我们将专注于研究复杂细胞环境中的蛋白质动力学。目前的固态NMR技术的灵敏度很差，因为它只能读出低伽马核（例如碳-13）的信息。这可以通过使用灵敏度增强技术，动态核极化（DNP）NMR来解决，但这必须在极冷的温度（100 K）下进行，这会消除动态运动。目前的合作项目是学习和应用快速MAS-NMR，在室温下进行，报告更高的伽马核（例如氢-1），以研究淀粉样蛋白原纤维在其天然细胞环境中的蛋白质动力学。该项目的更广泛影响包括将这些方法应用于研究由蛋白质形成的纤维，如与阿尔茨海默病有关的tau蛋白。该项目将包括一种平衡的方法，使用在国内实验室开发的创新样品制备方法和完善的方法和仪器，在浙江大学的EAPSI研究中证明生物分子NMR的灵敏度显着增强。这项工作将导致两个主要的发现i）如果快速MAS-NMR可以用于研究蛋白质在内源性水平和ii）确定细胞环境是否影响蛋白质动力学，特别是内在无序区域。所提出的研究的智力价值在于建立快速MAS-NMR作为DNP-NMR的补充技术，用于在复杂的细胞环境中研究内源性水平的蛋白质。其更广泛的影响在于开发替代方法，以进一步研究产生通常与疾病状态有关的纤维的内在无序蛋白质。该奖项是东亚和太平洋夏季研究所项目下的一项美国研究生夏季研究，由NSF和中国科技部共同资助。

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.