Structure and Dynamics of Proteins from NMR Orientational Data

建议继续探索溶液中蛋白质结构和动力学的潜在新信息来源。新的信息来自于在蛋白质的核磁共振光谱中看到的残留偶极耦合，当它们与各向异性介质（如水性液晶）相互作用或与足够高的磁场直接相互作用时，它们被诱导具有低水平的空间秩序。容易测量的耦合通过它们对平均相互作用向量长度和方向的依赖，为获取蛋白质结构和动力学信息提供了有效途径。除了对分离蛋白质的基本结构研究外，目标还包括：(i)开发和应用一种方法来定义溶液中蛋白质-蛋白质相互作用的几何形状，以及（ii）开发和应用一种策略来量化蛋白质内部运动的振幅。蛋白质-蛋白质相互作用的几何结构的定义有助于将结构和机制描述从孤立的蛋白质域转移到理解完整生命过程所需的系统范围描述。内部运动振幅的量化提供了对蛋白质功能的机械细节的深入了解，这些细节可能在结构的静态表示中被遗漏。用于测试该方法的目标系统包括许多特性良好的蛋白质，如肌红蛋白和红霉素，但也包括最近从嗜热生物中发现的红霉素的电子转移伙伴。就总体影响而言，该项目将为本科生、研究生和博士后提供培训机会。在后基因组时代，在分子水平上定义蛋白质功能的方法的开发和使用方法的培训变得越来越重要。大量的蛋白质序列信息可以对新的生物技术产品的开发和疾病的治疗产生影响，但只有在对蛋白质功能有充分的了解的情况下。

The continued exploration of a potential source of new information on protein structure and dynamics in solution is proposed. The new information comes from residual dipolar couplings seen in NMR spectra of proteins when they are induced to have low levels of spatial order by their interaction with an anisotropic medium, such as an aqueous liquid crystal, or by their direct interaction with a sufficiently high magnetic field. The easily measured couplings provide an efficient route to information about protein structure and dynamics through their dependence on average interaction vector length and orientation. In addition to basic structural studies of isolated proteins, objectives include, (i) the development and application of a method for defining the geometry of protein-protein interactions in solution, and (ii) the development and application of a strategy for quantifying the amplitudes of internal motions in proteins. The definition of the geometry of protein-protein interaction serves to move structural and mechanistic descriptions from isolated protein domains toward the system wide descriptions that are needed for understanding complete life processes. The quantification of amplitudes of internal motion provides insight into mechanistic details of protein function that might be missed in static representations of structure. The target systems for testing the methodology include a number of well characterized proteins such as myoglobin and rubredoxin, but also include recently identified electron transfer partners of a rubredoxin from a thermophilic organism.In terms of general impact, this project will provide training opportunities for undergraduate, graduate, and postdoctoral students. Development of methods and training in the use of methods that can define protein function at a molecular level is becoming increasingly important in a post-genomic era. Vast quantities of protein sequence information can have an impact on the development of new biotechnology products, and the treatment of disease, but only with an adequate understanding of protein function.