Pressure apparatus for nuclear magnetic resonance studies of protein excited states

我们要求资金能够在1巴和2000巴之间的压力进行核磁共振（NMR）研究。多伦多大学的这种独一无二的仪器将被安装在现有的蛋白质核磁共振设备中，供各种研究人员使用。温度和压力是大分子和蛋白质结构研究中非常有用的参数。我们的主要研究方向包括光开关蛋白和膜蛋白。在这两种情况下，我们都可以用基态和一个或多个低处激发态来描述蛋白质。激发态和激发态中间体通常与蛋白质的活性形式有关，并且通常是最难以理解的。一种叫做核磁共振色散光谱的技术的应用使我们能够梳理出与蛋白质及其激发态相关的能量格局。通过重复这些测量作为压力的函数，我们可以了解激发态是如何构建的基本信息。该装置使我们在目前的研究中取得了巨大的飞跃（例如激发态的比体积和可压缩性，折叠蛋白质的水渗透，关于光开关蛋白质的新信息），并通过高分辨率核磁共振研究压力依赖性现象。我们也相信，该机构将建立新的合作关系，并为申请人建立新的研究领域。

We request funds to be able to perform Nuclear Magnetic Resonance (NMR) studies at pressures between 1 bar and 2000 bar. The one-of-a-kind apparatus at the University of Toronto would be installed in an existing protein NMR facility and used by various researchers. Temperature and pressure are critically useful parameters in structure studies of macromolecules and proteins. Our main research interests include photo-switchable proteins and membrane proteins. In both cases, we may describe the proteins in terms of a ground state and one or more low-lying excited states. The excited states and excited state intermediates are often associated with the active form of the protein and are often the most elusive to understand. The application of a technique called NMR dispersion spectroscopy allows us to tease out the energy landscape associated with the protein and its excited state(s). By repeating these measurements as a function of pressure we can learn fundamental information about how the excited states are constructed. The apparatus permits us to make great leaps in our current research (e.g. specific volumes and compressibilities of excited states, water penetration of folded proteins, new information on photoswitchable proteins) and study pressure-dependent phenomena by high resolution NMR. We also believe that that the apparatus will forge new collaborations and establish new research niches for the applicants.