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

通过位点特异性二维红外光谱研究模块化氧化还原酶的构象和动力学

• 批准号: 2041692
• 负责人: Changjian Feng
• 金额: $ 65.15万
• 依托单位: University of New Mexico Health Sciences Center
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-07-01 至 2025-06-30
• 项目状态: 未结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=2041692&HistoricalAwards=false
• 关键词: Conformations; Dynamics; Modular; Redox; Enzymes

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.

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。这样一种授权技术目前在家庭组织中是缺乏的。由于新墨西哥州的大学是一个西班牙裔服务机构，这个奖项将提供一线的研究机会，很少提供给他们代表性不足的少数民族学生。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，以确保一支多元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理解电子转移潜在机制的最前沿的紧迫挑战，电子转移是生物学中的一个基本过程。这个项目的重点是一氧化氮合酶（NOS），大，模块酶的构象动力学。NOS是重要的，因为一氧化氮的产生在不同的信号传导过程中的关键作用。NOS的一个标志是其具有高度灵活的系链的多结构域结构，允许动态的、受调节的结构域间电子转移。NOS酶已被广泛研究，但由于内在的复杂性，不仅涉及动力学，而且还涉及伴侣蛋白结合和翻译后修饰，它是如何调节的一个全面的图片仍然缺乏。定量的洞察到大规模的域运动和更本地化的动态在停靠状态是必要的，以提供机械细节。这个过渡项目将使PI能够通过特定位置的二维红外（IR）光谱在研究对接状态构象和动力学方面开辟新的方向。当与IR探针的位点选择性标记所提供的空间精度相结合时，该方法将使具有高空间和时间分辨率的NOS对接状态的构象和动力学的前所未有的研究成为可能。这个多学科项目的目标是实现NOS对接状态的结构动态的特定残留特征，并描述它们对功能的贡献。由于NOS是一种范式信号系统，因此其结果将产生广泛的影响。所提出的方法也将提供一个蓝图，研究其他复杂的，动态的蛋白质系统，其功能是门控结构域之间的电子转移的构象控制。因此，该项目代表了研究多结构域蛋白质结构-动力学-功能关系的令人兴奋的下一步。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。

项目成果

Interdomain Interactions Modulate the Active Site Dynamics of Human Inducible Nitric Oxide Synthase.

• DOI: 10.1021/acs.jpcb.2c04091
• 发表时间: 2022-09-15
• 期刊: JOURNAL OF PHYSICAL CHEMISTRY B
• 影响因子: 3.3
• 作者: Tumbic, Goran W.;Li, Jinghui;Jiang, Ting;Hossan, Md Yeathad;Feng, Changjian;Thielges, Megan C.
• 通讯作者: Thielges, Megan C.

Probing calmodulin-NO synthase interactions via site-specific infrared spectroscopy: an introductory investigation

• DOI: 10.1007/s00775-024-02046-0
• 发表时间: 2024-04-05
• 期刊: JOURNAL OF BIOLOGICAL INORGANIC CHEMISTRY
• 影响因子: 3
• 作者: Singh，Swapnil;Gyawali，Yadav Prasad;Feng，Changjian
• 通讯作者: Feng，Changjian

Probing Protein Dynamics in Neuronal Nitric Oxide Synthase by Quantitative Cross-Linking Mass Spectrometry.

• DOI: 10.1021/acs.biochem.3c00245
• 发表时间: 2023-08-01
• 期刊: BIOCHEMISTRY
• 影响因子: 2.9
• 作者: Jiang, Ting;Wan, Guanghua;Zhang, Haikun;Gyawali, Yadav Prasad;Underbakke, Eric S.;Feng, Changjian
• 通讯作者: Feng, Changjian