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

职业：通过溶液态核磁共振研究蛋白质动力学的新实验

• 批准号: 0445643
• 负责人: Nikolai Skrynnikov
• 金额: --
• 依托单位: Purdue University
• 依托单位国家: 美国
• 项目类别: Continuing Grant
• 财政年份: 2005
• 资助国家: 美国
• 起止时间: 2005-03-01 至 2010-08-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=0445643&HistoricalAwards=false
• 关键词: CAREER; New; Experiments; Studying; Protein

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.

这个CAREER项目的目标是开发新的NMR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和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，将受益于对蛋白质运动的新见解。微秒毫秒的时间尺度动力学将使用掺杂有Ln 3+离子的蛋白质样品进行研究。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的直接结构表征。微秒-毫秒时间尺度的动力学将通过应用具有新的振幅（或相位）模式的射频场来探测。该技术将改进运动的采样，并提供若干技术优势（例如，关于样品加热和载体偏移效应）。皮秒-纳秒时间尺度的动力学将通过补充一套标准的15 N弛豫测量目标氘代酰胺网站，15 N-2 H。这些实验对于研究1纳秒时间尺度上的局部动力学（如部分折叠或未折叠蛋白质中的节段运动）特别有价值。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。作为这个职业生涯项目的核心部分，PI将设计和实施一个目标导向的方法来教学NMR光谱。目标将是教学生如何解决一个小蛋白质的结构，通过溶液状态NMR。本课程所涵盖的每一个理论或实验概念都将作为实现最终目标的特定步骤的一种手段。学生将参加一系列模拟现实生活中结构确定研究的实践练习。目标导向的教学将是非常有用的其他先进的光谱课程以外的领域的NMR。将作出特别努力，以满足不同的教育，社会和文化背景的学生的需求，特别是关于代表性不足的群体。将进行非正式的一对一咨询会议，以指导少数民族学生走向最佳的学习策略。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的学生加入他的研究小组。