Fluorescent Probes for Studying Copper Oxidation Biology

这个项目的长期目标是在分子水平上理解金属和金属之间的关系。 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。 这项工作的核心是跟踪生物体内氧化还原活性金属离子和氧代谢物的能力。 系统。为了满足这一需求，我们将设计铜和过氧化氢的荧光传感器，这是两种主要的 有助于体内的氧化信号和压力，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处理酶所必需的氧化还原活性辅因子， 抗氧化剂、结缔组织和神经递质的处理，但其细胞管理不善可能引发 无节制地产生过氧化氢和相关的活性氧所造成的氧化应激 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能导致 与年龄相关的严重人类疾病，包括癌症，组织和器官系统的功能衰退， 心血管疾病和神经退行性疾病，如阿尔茨海默病(AD)。过氧化氢含量更高 然而，作为一种混杂的ROS氧化剂，因为新出现的证据表明，这种受调控的生产 氧代谢产物在控制细胞增殖和/或细胞死亡的中介事件中起关键作用。这个 这里设计的化学传感器将为追踪特定的分子物种提供强大的新化学工具。 被怀疑参与氧化信号和/或应激途径。具体目标包括：(一)发展 活体细胞内不稳定铜(L)和铜(11)选择性灵敏荧光传感器的研制 用于H_2O_2选择性活细胞成像的类似小分子试剂，(III)及其应用于 研究铜和过氧化氢在健康和疾病细胞中的积累、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的相互作用 生命系统与理解它们对严重的公共卫生疾病的贡献有关。这些 对于年龄是风险因素的疾病，包括阿尔茨海默氏症和相关疾病，努力尤为重要 神经退行性疾病，因为他们有很高的金属调节失调和氧化应激的发生率。

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.