Molecular Mechanisms Underlying Mammalian NADPH Oxidase Activation and Regulation

项目总结 维持活性氧(ROS)的动态平衡对于保持细胞的完整性和生命力至关重要 为了几乎所有生命的生存和成长。在多细胞生物中，ROS是在细胞外活跃地产生的 细胞或靠近细胞膜，以防止入侵的病原体，以及在正常的生理条件下 荷尔蒙生物合成等过程。然而，ROS通常与对 细胞内的蛋白质和DNA。过量的ROS产生会导致氧化应激，并导致 许多慢性疾病的发展，如衰老、癌症、糖尿病、心脏疾病和 神经退行性疾病。NADPH氧化酶，一个膜酶家族，其主要功能是 以产生ROS，在维持ROS动态平衡方面发挥重要作用，从而成为有效的药物靶点 用于对抗与氧化应激有关的多种疾病。NADPH氧化酶通过以下途径产生ROS 催化跨膜电子从胞内NADPH到胞外氧的转移。哺乳动物编码 7种NADPH氧化酶：DUOX1-2和NOX1-5。到目前为止，人们对其分子机制知之甚少。 管理NADPH氧化酶蛋白的激活和调节，这是一个关键的知识鸿沟。在……里面 在这项建议中，将建立一个跨学科的研究计划，以研究 NADPH氧化酶：结合尖端结构生物学技术，如单粒子低温EM和 生物化学、生物物理学和细胞生物学方法。我们的目标是解决两个基本问题 NADPH氧化酶催化活性的基础：I)NADPH氧化酶如何介导跨膜 电子转移催化ROS的产生？II)NADPH的催化作用是怎样的 氧化物酶在分子水平上被激活和调节？以DUOX1为例，我们将 建立了解NADPH氧化酶结构与功能关系的分子范式。这个 我们的研究结果将促进我们对NADPH氧化酶生物学和产卵的基本理解 为对抗氧化应激的新药开发战略奠定了基础。

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.