Tools for Probing Conformational Dynamics of Membrane Proteins

生物分子的运动被称为动力学，是许多生物功能的必要条件，包括酶催化、分子识别、蛋白质-蛋白质相互作用和底物运输。虽然利用x射线晶体学在确定膜蛋白离散构象的结构方面取得了进展，但对蛋白质动力学在功能中的作用知之甚少。本项目的总体目标是开发新的实验方法来灵敏地测量膜蛋白所需的功能相关运动。该项目将吸引高中生、本科生和研究生参与研究，进一步传播生物物理化学、化学生物学、生物化学和结构生物学等跨学科领域的知识，培养面向生物产业和经济的STEM人才。本课题将重点研究定向样品核磁共振（OS-NMR）实验的开发与应用。这些方法将应用于三种不同大小和复杂程度的膜蛋白转运体，以确定这些方法揭示功能重要的膜蛋白动力学的可行性。该项目将开发方法来克服研究膜蛋白动力学的主要挑战。具体而言，将开发涉及底物运输的膜蛋白构象运动的生物物理学见解和固态核磁共振技术开发中的光谱创新，以探测动力学。改善50多个成员院校的本科课程和科学培训。该项目得到了生物科学理事会分子和细胞生物科学部分子生物物理集群的支持。该奖项反映了美国国家科学基金会的法定使命，并通过使用基金会的知识价值和更广泛的影响审查标准进行评估，被认为值得支持。

Motion in biological molecules, termed dynamics, is a requirement for many biological functions including enzyme catalysis, molecular recognition, protein-protein interactions, and substrate transport. While progress has been made to determine structures of membrane proteins in discrete conformations using X-ray crystallography, much less is known about the role of protein dynamics in function. The overall goal of this project is to develop novel experimental methods to sensitively measure functionally-relevant motions required of membrane proteins. This project will involve high school, undergraduate, and graduate students in research, which will further disseminate knowledge in the interdisciplinary fields of biophysical chemistry, chemical biology, biochemistry, and structural biology and develop STEM workforce for bio-industry and -economy. This research will focus on the development and application of oriented sample nuclear magnetic resonance (OS-NMR) experiments. These methods will be applied to three membrane protein transporters of varying size and complexity to establish the feasibility of the methods to reveal functionally important membrane protein dynamics. This project will develop methods to overcome the major challenges in studying membrane protein dynamics. Specifically, biophysical insight into conformational motions of membrane proteins involved in substrate transport and spectroscopic innovation in the development of solid-state NMR techniques to probe dynamics will be developed. to improve undergraduate curricula and scientific training across the 50+ member institutions. This project is supported by the Molecular Biophysics Cluster of the Molecular and Cellular Biosciences Division in the Biological Sciences Directorate.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.