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

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.