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

蛋白质需要移动才能发挥作用，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一种现代研究工具，以确定蛋白质在执行特定功能时如何移动。目前家庭组织缺乏这种授权技术。由于新墨西哥大学是一所西班牙裔服务机构，该奖项将提供前沿研究机会，而这些机会很少提供给其代表性不足的少数族裔学生。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，以确保多元化的科学队伍继续从事积极的前沿研究。在了解电子转移（生物学的基本过程）的潜在机制方面，破译蛋白质动力学是一项紧迫的挑战。该项目重点研究一氧化氮合酶 (NOS)（大型模块化酶）的构象动力学。 NOS 很重要，因为一氧化氮的产生在多种信号传导过程中发挥着关键作用。 NOS 的标志之一是其多域架构和高度灵活的系链，允许动态、受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，因此仍然缺乏对其调控方式的全面了解。为了提供机制细节，有必要对大规模域运动和对接状态下更局部的动态进行定量分析。该转变项目将使 PI 能够通过特定位点的二维红外 (IR) 光谱研究对接态构象和动力学开辟新方向。当与红外探针位点选择性标记提供的空间精度相结合时，该方法将能够以高空间和时间分辨率对 NOS 对接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 对接态结构动力学的残基特异性表征，并描述它们对功能的贡献。由于 NOS 是一个范例信号系统，其结果将产生广泛的影响。所提出的方法还将为研究其他复杂的动态蛋白质系统提供蓝图，其中其功能是通过域之间电子转移的构象控制来门控的。因此，该项目代表了研究多域蛋白质的结构-动力学-功能关系的一个令人兴奋的下一步。该奖项反映了 NSF 的法定使命，并通过使用基金会的智力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.