New methods development for structure determination of multihelical membrane proteins in their native-like lipid bilayers by solid-state NMR with applications

在这个由化学部化学测量和成像计划资助的项目中，亚历山大·涅夫佐罗夫教授正在通过固态核磁共振（NMR）光谱研究膜蛋白（MP）在其天然条件下的性质。膜蛋白执行过多的生物学功能，但其解决的结构的数量仍然相对较少。固态NMR技术使研究人员能够对在接近生理条件下难以或不可能结晶的MP进行结构-功能研究。 在本研究中开发的NMR光谱方法将该技术的能力扩展到多螺旋MPs.The主要目的是通过开发新的光谱工具，包括距离测量和拥挤光谱区域分配的方法，来改变定向膜蛋白（MPs）的固态NMR。此外，正在创建基于纳米孔和定向大圆盘的柔性脂质组合物的新膜模拟物以监测脂质诱导的MP构象变化。探讨了脂双层的动力学性质，包括膜包埋蛋白质的旋转扩散和双层粘度。潜在的未来应用包括参与阿尔茨海默病和其他神经系统疾病的人类乙酰胆碱受体的结构测定。由于该项目的广泛影响，该研究的高度跨学科性质为参与的年轻科学家提供了多方面的培训。作为外展活动，通过凯南研究员的指导，Nevzorov博士将他的研究成果推广到高中教育和STEM项目。

In this Project funded by the Chemical Measurement and Imaging program of the Chemistry Division, Professor Alexander Nevzorov is studying membrane proteins (MPs) under their native-like conditions by solid-state Nuclear Magnetic Resonance (NMR) spectroscopy. Membrane proteins perform a plethora of biological functions, yet the number of their solved structures remains relatively small. The solid-state NMR technique allows researchers to perform structure-function studies of MPs that are difficult or impossible to crystallize at nearly physiological conditions. NMR spectroscopic methods developed during this research expand capabilities of this technique to multihelical MPs.The main purpose of this project is to transform solid-state NMR of oriented membrane proteins (MPs) by developing new spectroscopic tools including methods for distance measurements and assignment of crowded spectral regions. Moreover, new membrane mimetics of flexible lipid composition based on nanopores and oriented macrodisks are being created to monitor lipid-induced conformational changes of MPs. Dynamic properties of lipid bilayers are probed including rotational diffusion of membrane-embedded proteins and the bilayer viscosity. Potential future applications include structure determination of the human Acetylcholine receptor involved in the Alzheimer's disease and other neurological disorders. As broader impacts of this project, the highly interdisciplinary nature of this research provides versatile training for the young scientists involved. As outreach activities, via mentorship of a Kenan Fellow, Dr. Nevzorov promotes the results of his research to high-school education and STEM programs.