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

本CAREER项目的目标是开发新的核磁共振弛豫方法，以获得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与生物功能之间的重要关系。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。将使用掺杂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的数量，更重要的是，允许对许多蛋白质的生物功能至关重要的瞬时构象物种进行直接的结构表征。微秒-毫秒时间尺度动力学将通过应用新的振幅（或相位）模式的射频场来探测。这种技术将改善运动的采样，并提供几个技术优势（例如，关于样品加热和载流子偏移效应）。皮秒-纳秒时间尺度动力学将通过补充针对氘化酰胺位点15N- 2h的标准15N弛豫测量集进行研究。这些实验对于研究约1纳秒时间尺度的局部动力学，如部分折叠或未折叠蛋白质的片段运动，将特别有价值。虽然研究工具的开发非常重要，但教育和培训将应用这些工具的未来研究人员同样重要。作为CAREER项目的核心部分，PI将设计并实施一种目标导向的核磁共振波谱学教学方法。我们的目标是教学生如何通过溶液态核磁共振来解析小蛋白质的结构。讲座课程中涉及的每一个理论或实验概念都将作为实现最终目标的某一特定步骤的手段来介绍。学生将参与一系列模拟现实生活中的结构确定研究的实践练习。这种目标导向的教学方法对核磁共振以外的其他高级光谱学课程有很大的借鉴意义。将作出特别努力，满足具有不同教育、社会和文化背景的学生的需要，特别是对代表性不足的群体的需要。非正式的一对一咨询会议将进行，以指导少数民族学生的最佳学习策略。PI将通过课堂互动和与少数民族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.