CAREER: Oligomeric Stoichiometry of Superoxide Dismutase 1 and Neuronal Antioxidant Defense

高水平的代谢活动要求神经元有能力处理氧化应激。该项目旨在了解神经元在遇到环境氧化试剂时如何调节一线抗氧化蛋白-铜/锌超氧化物歧化酶(SOD1)的组装。该项目的结果将揭示氧化应激如何改变神经元抗氧化防御的SOD1结构。此外，PI还将启动一项新的教育计划--“定量生物物理理解”(Qbic)，教学生如何使用生物成像实验来量化蛋白质的结构状态。将特别重视与休斯顿大学学生分会合作，有效扩大妇女和代表不足的少数群体在STEM中的参与。QBIC将提供跨学科培训，现代化的化学课程，并为NSF发展强大的STEM工作力量的目标做出贡献。SOD1寡聚体状态的调节在其功能中发挥关键作用，因此对神经元抗氧化防御至关重要；然而，由于缺乏适当的分析工具来量化细胞中的蛋白质寡聚，其分子机制尚不清楚。在这项研究计划的第一部分，PI将开发一种定量的单分子方法来量化细胞核、胞浆和线粒体中的SOD1寡聚。细胞内的氧化还原传感器将被用来定量亚细胞的氧化状态，质谱学将被用来定量不同环境应激源下SOD1翻译后修饰的变化。这项工作的成功将首次回答SOD1相互作用和寡聚体分布是如何在神经元中对环境刺激做出精细调整的。这一奖项反映了NSF的法定使命，并通过使用基金会的智力优势和更广泛的影响审查标准进行评估，被认为值得支持。

High levels of metabolic activity require neurons to be competent in handling oxidative stress. This project seeks to understand how neurons modulate the assembly of Cu/Zn superoxide dismutase (SOD1), the frontline antioxidant protein, when they encounter environmental oxidative reagents. The results of this project will inform how oxidative stress changes SOD1 structures for neuronal antioxidant defense. In addition, the PI will initiate a new educational program, 'Quantitative Biophysical Comprehension' (QBiC), to teach students how to quantify the structural state of a protein using bio-imaging experiments. Particular emphasis will be placed on working with University of Houston student chapters to effectively broaden the participation of women and underrepresented minorities in STEM. QBiC will provide interdisciplinary training, modernize the chemistry curriculum and contribute to the NSF goal of developing a strong STEM workforce.Modulation of SOD1 oligomeric states plays a critical role in its function and is thus essential to neuronal antioxidant defense; however, its molecular mechanisms are poorly understood due to the lack of proper analytical tools to quantify protein oligomerization in cells. In the first part of this research program, the PI will develop a quantitative single-molecule approach to quantify SOD1 oligomerizations in the nucleus, cytosol, and mitochondria. An in-cell redox sensor will be employed to quantify the subcellular oxidative status, and mass spectrometry will be used to quantify changes in SOD1 posttranslational modifications under various environmental stressors. The success of this work will answer for the first time how SOD1 interactions and oligomeric distribution are finely tuned in neurons in response to environmental stimuli.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.