Molecular Mechanisms Underlying Mammalian NADPH Oxidase Activation and Regulation

项目摘要 维持活性氧（ROS）稳态对于保持细胞完整性和生命力至关重要。 几乎所有生命的生存和成长。在多细胞生物中，ROS在细胞外活跃地产生。 细胞或细胞膜附近，以保护免受入侵的病原体以及在正常的生理 如激素生物合成过程。然而，ROS通常与引起损伤有关， 细胞内的蛋白质和DNA。过量的ROS产生导致氧化应激，并有助于 许多慢性疾病的发展，如衰老、癌症、糖尿病、心脏病和 神经退行性疾病NADPH氧化酶是膜酶家族，其主要功能是 产生活性氧，在维持活性氧稳态中起重要作用，因此可作为有效的药物靶点 用于对抗与氧化应激相关的多种疾病。NADPH氧化酶通过以下途径产生ROS： 催化从胞质NADPH到细胞外氧的跨膜电子转移。哺乳动物编码 七种NADPH氧化酶：DUOX 1 -2和NOX 1 -5。到目前为止，人们对这一分子机制知之甚少， 控制NADPH氧化酶蛋白的激活和调节，代表了一个关键的知识空白。在 为此，将建立一个跨学科的研究计划，研究 NADPH氧化酶通过结合尖端的结构生物学技术，如单粒子cryoEM与 生物化学、生物物理和细胞生物学方法。我们的目标是解决两个基本问题 NADPH氧化酶的催化活性的基础：i）NADPH氧化酶如何介导跨膜 电子转移催化活性氧的产生？NADPH的催化功能如何 在分子水平上激活和调节氧化酶？以DUOX 1为例，我们将 为理解NADPH氧化酶的结构-功能关系建立了分子范式。的 我们的研究成果将促进我们对NADPH氧化酶生物学的基本理解， 对抗氧化应激的新药开发策略的基础。

PROJECT SUMMARY Maintenance of the reactive oxygen species (ROS) homeostasis is essential to preserve cell integrity and vital for the survival and growth of almost all life. In multicellular organisms, ROS is actively generated outside of the cell or near the cell membrane to protect against invading pathogens as well as in normal physiological processes such as hormone biosynthesis. However, ROS are generally associated with causing damage to proteins and DNA within cells. Excessive ROS production leads to oxidative stress and contributes to the development of many chronic conditions such as aging, cancer, diabetes, cardiac disorders, and neurodegenerative diseases. The NADPH oxidases, a family of membrane enzymes whose primary function is to produce ROS, play an essential role in maintaining ROS homeostasis and thus serve as valid drug targets for combatting numerous diseases associated with oxidative stress. NADPH oxidases generate ROS by catalyzing cross-membrane electron transfer from cytosolic NADPH to extracellular oxygen. Mammals encode seven NADPH oxidases: DUOX1-2 and NOX1-5. To date, little is known about the molecular mechanism governing the activation and regulation of NADPH oxidase proteins, representing a critical knowledge gap. In this proposal, an interdisciplinary research program will be established to study the working mechanism of NADPH oxidases by combining cutting-edge structural biology techniques such as single-particle cryoEM with biochemical, biophysical, and cell biology approaches. We aim to address the two fundamental questions underlying the catalytic activity of NADPH oxidases: i) how do NADPH oxidases mediate cross-membrane electron transfer to catalyze the production of ROS? And ii) how is the catalytic function of NADPH oxidases activated and regulated at the molecular level? Using the DUOX1 as an example, we will establish a molecular paradigm for understanding the structure-function relationship of NADPH oxidases. The outcomes of our studies will advance our fundamental understanding of the NADPH oxidase biology and lay the foundation for novel drug development strategies to combat oxidative stress.