CAREER: New Experiments for Studying Protein Dynamics by Solution-state NMR

这个职业项目的目标是开发新的核磁共振弛豫方法，以获得关于溶液中蛋白质动力学的新信息。最终，这些研究将有助于阐明蛋白质动力学和生物功能之间至关重要的关系。许多领域，如蛋白质工程和蛋白质的计算机建模，都将受益于对蛋白质运动的新见解。我们将使用掺入Ln3离子的蛋白质样品来研究微秒-毫秒时间尺度的动力学。这种方法将增加报告局部运动的自旋数量，更重要的是，允许直接表征对许多蛋白质的生物功能至关重要的瞬时构象物种的结构。微秒-毫秒时间尺度的动力学将通过应用具有新的幅度(或相位)模式的射频场来探测。这项技术将改进运动的采样，并提供几个技术优势(例如，在样品加热和载波补偿效果方面)。皮秒-纳秒的时间尺度动力学将通过补充一组标准的15N弛豫测量来研究，该测量目标是15N-2H。这些实验对于在1纳秒时间尺度上研究局部动力学特别有价值，例如部分折叠或展开蛋白质的节段性运动。虽然研究工具的开发非常重要，但教育和培训未来将应用这些工具的研究人员也同样重要。作为这一职业项目的核心部分，PI将设计和实施一种面向目标的方法来教授核磁共振光谱学。目标是教学生如何通过溶液状态核磁共振来解决小蛋白质的结构问题。课程中所涵盖的每个理论或实验概念都将被介绍为实现最终目标的特定步骤的一种手段。学生们将参加一系列的实践练习，模拟现实生活中的结构确定研究。这种以目标为导向的教学将对核磁共振以外的其他高级光谱学课程非常有用。将特别努力满足具有不同教育、社会和文化背景的学生的需要，特别是在代表人数不足的群体方面。将进行非正式的一对一咨询会议，以指导少数族裔学生采取最佳学习策略。国际学生联合会将通过课堂互动和与少数族裔本科院校的积极接触，积极从代表性不足的群体中招募更多学生加入他的研究小组。

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.