Methodology for Rapid and Precise Measurement of NMR Parameters: Development and Application to Study Protein Structure and Folding

核磁共振波谱学在科学和工程中有着广泛的应用，但由于最小测量次数随维数的急剧增加而导致的核磁共振采样问题阻碍了多维光谱信息的获取。此外，基于核磁共振的生物大分子研究受到化学位移简并的阻碍，部分原因是位移测量的精度相对较低。该项目的重点是开发快速核磁共振数据采集方法和数据采集方案，使人们能够更准确地测量核磁共振参数。具体而言，目标是实现一条新的“同时循环”或“切片选择”核磁共振实验线，以（准）同时执行翻转角循环的射频脉冲相位，并实现不含色散线分量的光谱获取方案，这些色散线分量妨碍准确测量位移。这种新方法将应用于解决结构基因组学联盟框架中的蛋白质结构，以评估对半自动数据分析和蛋白质折叠的影响。这种新的核磁共振方法将对科学界核磁共振数据采集的速度和准确性产生广泛的影响。民意调查将设立一个门户网站，以确保有效地传播结果。该项目非常适合培养基于核磁共振的结构生物学的年轻研究人员，特别是在核磁共振研究蛋白质结构和折叠的高级方法开发和应用方面。本研究将结合纽约州立大学布法罗分校化学系和结构生物系青年科学家跨学科综合教学的具体需求进行。PI将提供物理化学和核磁共振在生命科学中的应用的本科和研究生水平的课程，他将继续支持本科生在实验室、暑期学校和大学的研究培训。该项目由生物科学理事会分子与细胞生物科学部的分子生物物理学和数学与物理科学理事会化学部的实验物理化学项目共同支持。

NMR spectroscopy is widely used in science and engineering, but acquisition of multi-dimensional spectral information is hampered by the NMR sampling problem resulting from the steep increase of minimal measurement times with dimensionality. Furthermore, NMR-based studies of biological macromolecules are impeded by chemical shift degeneracy, in part resulting from comparably low accuracy of shift measurements. The focus of this project is on the development of methodology for rapid NMR data acquisition and data acquisition schemes enabling one to measure NMR parameters more accurately. Specifically, the objective is to implement a new line of "simultaneously cycled" or "slice selected" NMR experiments to (quasi) simultaneously execute radio-frequency pulse phase of flip-angle cycling, and to implement schemes to acquire spectra devoid of dispersive line components, which impede accurate measurement of shifts. This new methodology will be applied to solve protein structures in the framework of structural genomics consortia in order to assess impact on semi-automated data analysis and protein folding. This new NMR methodology will broadly impact on the speed and accuracy of NMR data acquisition in the scientific community. The PI will set up a web portal to ensure efficient dissemination of results. This project is well suited to educate young researchers in NMR-based structural biology, in particular in advanced methodology development and application of NMR to study protein structure and folding. The research will be conducted considering the specific demands for comprehensive interdisciplinary teaching of young scientists at the Departments of Chemistry and Structural Biology at the State University at New York at Buffalo. The PI will offer undergraduate and graduate level courses in Physical Chemistry and on the use of NMR in life sciences, and he will continue to support research training of undergraduate students in the laboratory, in summer schools and in colleges. This project is jointly supported by Molecular Biophysics in the Division of Molecular and Cellular Biosciences in the Directorate for Biological Sciences and the Experimental Physical Chemistry Program in the Division of Chemistry in the Mathematical and Physical Sciences Directorate.