The objective of this CAREER project is to develop new NMR relaxation methods to obtain new information on protein dynamics in solution. Ultimately, these studies will help to elucidate the all-important relationship between protein dynamics and biological function. A number of fields, such as protein engineering and computer modeling of proteins, stand to benefit from new insights into protein motions. Microsecond-millisecond time-scale dynamics will be investigated using protein samples doped with Ln3+ ions. This approach will increase the number of spins reporting on local motions and, more importantly, permit direct structural characterization of transient conformational species which are critical for biological function of many proteins. Microsecond-millisecond time-scale dynamics will be probed by application of rf fields with new amplitude (or phase) patterns. This technique will improve the sampling of motion and offer several technical advantages (e.g. with regard to sample heating and carrier offset effects). Picosecond-nanosecond time-scale dynamics will be investigated by complementing the set of standard 15N relaxation measurements targeting deuterated amide sites, 15N-2H. These experiments will be particularly valuable for studying local dynamics on a ~1 nanosecond time scale such as segmental motions in partially folded or unfolded proteins.
While the development of research tools is very important, it is equally important to educate and train future researchers who will be applying these tools. As a core part of this CAREER project, the PI will design and implement a goal-oriented approach to teaching NMR spectroscopy. The goal will be to teach students how to solve the structure of a small protein by means of solution-state NMR. Each theoretical or experimental concept covered in the lecture course will be introduced as a means to accomplish certain specific step toward the final goal. The students will participate in a series of practical exercises modeling a real-life structure determination study. The goal-oriented teaching will be very useful for other advanced spectroscopy courses outside the realm of NMR. A special effort will be made to meet the needs of students with diverse educational, societal, and cultural backgrounds, in particular with regard to underrepresented groups. Informal one-on-one counseling sessions will be conducted in order to direct minority students toward an optimal learning strategy. The PI will aggressively recruit more students from underrepresented groups to his research group, both through classroom interaction and active contacts with minority undergraduate institutions.
