Fluorescent Probes for Studying Copper Oxidation Biology

该计划的长期目标是在分子水平上了解金属之间的关系， 离子，氧代谢和氧化应激，以及它们对人类健康，衰老和疾病的贡献。 这项工作的核心是能够跟踪生物体内的氧化还原活性金属离子和氧代谢产物， 系统.为了满足这一需求，我们将设计用于铜和过氧化氢的荧光传感器，这两个主要的传感器 贡献者的氧化信号和压力在体内，并应用这些新的化学工具来研究 铜氧化生物学在神经退行性疾病和年龄相关疾病遗传模型中的作用。铜 是许多调节呼吸的氧加工酶所需的氧化还原活性辅因子， 抗氧化剂，结缔组织和神经递质处理，但其细胞管理不善可能引发 通过过氧化氢和相关活性氧的不受调节的产生产生的氧化应激 物种（ROS）。随着时间的推移，随后对蛋白质、脂质和核酸的氧化损伤可导致 在与年龄有关的严重人类疾病中，包括癌症， 心血管疾病和神经退行性疾病如阿尔茨海默病（AD）。H2 O2更多 而不是混杂的ROS氧化剂，然而，新出现的证据表明， 氧代谢物对于介导控制细胞增殖和/或细胞死亡的事件是关键的。的 本发明的化学传感器将为追踪特定的分子种类提供强有力的新化学工具 怀疑有助于氧化信号传导和/或应激途径。具体目标包括：（一）发展 用于成像活细胞中的不稳定Cu（I）和Cu（II）的选择性和灵敏的荧光传感器，（ii）设计 用于H2 O2的选择性活细胞成像的类似小分子试剂，（iii）以及将这些工具应用于 研究铜和过氧化物在健康和患病细胞中积累、运输和氧化还原功能， 组织模型研究金属营养素及其与氧代谢产物相互作用的基础科学， 生命系统与了解它们对严重公共卫生疾病的贡献有关。这些 努力对于年龄是风险因素的疾病特别重要，包括阿尔茨海默氏症和相关疾病。 神经退行性疾病，因为他们有金属失调和氧化应激的高发病率。

The long-term goal of this program is to understand, at the molecular level, the relationships between metal ions, oxygen metabolism, and oxidative stress, and their contributions to human health, aging, and disease. Central to this effort is the ability to track redox-active metal ions and oxygen metabolites in living biological systems. To meet this need, we will devise fluorescent sensors for copper and hydrogen peroxide, two major contributors to oxidative signaling and stress in the body, and apply these new chemical tools to study the roles of copper oxidation biology in genetic models of neurodegenerative and age-related diseases. Copper is a required redox-active cofactor for many oxygen-processing enzymes that regulate respiration, antioxidants, connective tissue, and neurotransmitter processing, but its cellular mismanagement can trigger oxidative stress through the unregulated production of hydrogen peroxide and related reactive oxygen species (ROS). Over time, subsequent oxidative damage to proteins, lipids, and nucleic acids can lead to the functional decline of tissue and organ systems in serious age-related human diseases, including cancer, cardiovascular disorders, and neurodegenerative diseases such as Alzheimer's disease (AD). H2O2 is more than a promiscuous ROS oxidant, however, as emerging evidence suggests that regulated production of this oxygen metabolite is critical for mediating events that control cell proliferation and/or cell death. The chemosensors devised here will provide powerful new chemical tools for tracking specific molecular species suspected of contributing to oxidative signaling and/or stress pathways. Specific aims include (i) developing selective and sensitive fluorescent sensors for imaging labile Cu(l) and Cu(ll) in living cells, (ii) devising analogous small-molecule reagents for selective live-cell imaging of H2O2, (iii) and applying these tools to study copper and peroxide accumulation, trafficking, and redox function in healthy and diseased cell and tissue models. Studying the basic science of metal nutrients and their interaction with oxygen metabolites in living systems is pertinent to understanding their contributions to serious diseases of public health. These efforts are of particular importance to diseases where age is a risk factor, including Alzheimer's and related neurodegenerative disorders, because they have high incidence of metal misregulation and oxidative stress.