Conformations and Dynamics of Modular Redox Enzymes via Site-Specific 2D Infrared Spectroscopy

蛋白质需要运动才能发挥作用，但科学家对这种运动仍然知之甚少。这项研究将使冯博士能够在自己的实验室中开发出一种现代研究工具，以确定蛋白质在执行特定功能时是如何移动的。这种授权技术目前在家庭组织中是缺乏的。由于新墨西哥大学是一所西班牙裔服务机构，该奖项将为他们代表性不足的少数族裔学生提供很少获得的一线研究机会。该项目还将为来自其他机构的优秀学生提供独特的研究机会，以确保多样化的科学队伍继续从事积极的前沿研究。破译蛋白质动力学是一个紧迫的挑战，在努力了解电子转移的潜在机制，在生物学的基本过程的最前沿。这个项目的重点是一氧化氮合酶（NOS）的构象动力学，大的，模块化的酶。一氧化氮之所以重要，是因为一氧化氮的产生在多种信号传导过程中起着关键作用。NOS的一个特点是它的多域架构具有高度灵活的系绳，允许动态，可调节的域间电子转移。NOS酶已被广泛研究，但由于其内在的复杂性，不仅涉及动力学，还涉及伴侣蛋白结合和翻译后修饰，因此仍缺乏对其如何调节的全面了解。对大尺度域运动和停靠状态下更局部的动力学进行定量分析对于提供机理细节是必要的。这个过渡项目将使PI在通过特定位置的二维红外光谱研究停靠态构象和动力学方面开辟新的方向。当结合红外探针提供的位置选择性标记的空间精度时，该方法将以高空间和时间分辨率对NOS停靠状态的构象和动力学进行前所未有的研究。这个多学科项目的目标是实现NOS对接状态结构动力学的残留特异性特征，并描述它们对功能的贡献。由于NOS是一个范式信号系统，研究结果将产生广泛的影响。所提出的方法也将为研究其他复杂的动态蛋白质系统提供蓝图，在这些系统中，其功能是通过结构域之间电子转移的构象控制来控制的。因此，该项目代表了研究多结构域蛋白结构-动力学-功能关系的一个令人兴奋的下一步。该奖项反映了美国国家科学基金会的法定使命，并通过使用基金会的知识价值和更广泛的影响审查标准进行评估，被认为值得支持。

Proteins need to move in order to function, but scientists still know very little about such motions. This research will allow Dr. Feng to develop a modern research tool in his own laboratory to identify how proteins move when performing specific functions. Such an empowering technique is currently lacking at the home organization. Since the University of New Mexico is a Hispanic Serving Institution, this award will provide frontline research opportunities that are rarely made available to their underrepresented minority students. The project will also provide unique research opportunities to outstanding students from other institutions, to ensure a diverse scientific workforce that remains engaged in active, cutting-edge research.Deciphering protein dynamics is a pressing challenge at the forefront of efforts to understand the underlying mechanisms of electron transfer, a fundamental process in biology. This project focuses on conformational dynamics of nitric oxide synthases (NOS), large, modular enzymes. NOSs are important because of the pivotal roles of nitric oxide production in diverse signaling processes. One hallmark of NOS is its multi-domain architecture with highly flexible tethers, allowing for dynamic, regulated interdomain electron transfer. The NOS enzyme has been extensively investigated, but due to an intrinsic complexity, involving not only dynamics, but also partner protein binding and posttranslational modification, a comprehensive picture of how it is regulated remains lacking. Quantitative insights into both the large-scale domain movements and the more localized dynamics in the docked state are necessary to provide mechanistic details. This Transitions project will enable the PI to forge a new direction in studying the docked state conformations and dynamics via site-specific 2D infrared (IR) spectroscopy. When combined with the spatial precision provided by site-selective labeling with IR probes, the approach will enable unprecedented study of the conformations and dynamics in the NOS docked states with high spatial and temporal resolution. The objective of this multidisciplinary project is to achieve residue-specific characterizations of structural dynamics of the NOS docked states and delineate their contributions to function. As NOS is a paradigm signaling system, the results will have a broad-reaching impact. The proposed approach will also provide a blueprint for studying other complex, dynamic protein systems in which its function is gated by conformational control of electron transfer between domains. This project thus represents an exciting next step in studying structure-dynamics-function relationships in multidomain proteins.This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.