Fluorescent Copper Probes for Intracellular Imaging

用于细胞内成像的荧光铜探针

• 批准号: 7217355
• 负责人: CHRISTOPH J FAHRNI
• 金额: $ 18.04万
• 依托单位: GEORGIA INSTITUTE OF TECHNOLOGY
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-04-01 至 2009-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7217355
• 关键词: ATP phosphohydrolase; Affinity; Alzheimer' s Disease; Amines; Amyotrophic Lateral Sclerosis; Architecture; Binding; Binding Proteins; Biological; Buffers; Cell Line; Cell Membrane Permeability; Cell membrane; Cells; Cellular Structures; Chelating Agents; Collaborations; Copper; Cytoplasm; DNA; Development; Disease; Electron Transport; Equilibrium; Evaluation; Fluorescence; Fluorescent Probes; Goals; Golgi Apparatus; Health; Human; Image; Ions; Iron; Kinetics; Laboratories; Lead; Libraries; Ligands; Link; Lipids; Localized; Measurement; Membrane Proteins; Menkes Kinky Hair Syndrome; Metabolism; Metals; Molecular; Monitor; Nature; Oxidation-Reduction; Process; Production; Property; Proteins; Reaction; Research Personnel; Respiration; Secretory Vesicles; Site; Solutions; Superoxides; Techniques; Therapeutic; Thermodynamics; Time; Toxic effect; Trace Elements; Universities; Washington; base; cofactor; extracellular; fluorophore; human disease; imaging probe; in vivo; nervous system disorder; novel diagnostics; programs; receptor; research study; tool; trafficking; uptake

DESCRIPTION (provided by applicant): Copper is an essential trace element, which is critical to human health. The redox properties of copper require a sophisticated management to avoid oxidative damage of lipids, proteins or DNA. Therefore the cell must maintain a subtle balance, such that copper is available for catalytic processes, but at the same time avoid accumulation to toxic levels. The detailed mechanisms and cellular structures responsible for maintaining control of intracellular copper levels are not well understood. This information is critical for the understanding of the molecular mechanisms of diseases caused by copper imbalance, such as Wilson's or Menkes' disease, both of which are fatal. More recently, certain neurological disorders including ALS and Alzheimer's disease have been linked to disorders in copper metabolism. One of the central hypotheses of the copper regulatory machinery suggests the presence of intracellular storage or buffer sites as a defense against deficiency, but also to protect the cell from abnormally high concentrations of copper. Golgi secretory vesicles which contain a copper ATPase membrane protein might function as intracellular compartments for copper storage. Fluorescence probes, which can permeate the plasma membrane are powerful tools for the study of intracellular metal ion distributions, yet rigorous analytical techniques for sensitive in vivo measurements of intracellular copper levels are completely lacking. The goal of the proposed project is the development of a copper specific fluorescent probe, which would not only allow to monitor vesicular labile copper concentrations, but would also provide information about the subcellular distribution in cell lines with disorders in copper metabolism. In a larger context a copper specific fluorescent probe will be of great importance for the long-term development of novel diagnostic and therapeutic tools for copper related human diseases.

说明(申请人提供)：铜是人体必需的微量元素，对人体健康至关重要。铜的氧化还原特性需要复杂的管理，以避免脂质、蛋白质或DNA的氧化损伤。因此，电池必须保持微妙的平衡，以便铜可用于催化过程，同时避免积累到有毒的水平。负责维持细胞内铜水平控制的详细机制和细胞结构尚不清楚。这些信息对于理解铜失衡引起的疾病的分子机制至关重要，例如威尔逊病或门克斯病，这两种疾病都是致命的。最近，包括肌萎缩侧索硬化症和阿尔茨海默病在内的某些神经系统疾病被认为与铜代谢障碍有关。 铜调节机制的中心假说之一表明，细胞内存储或缓冲位置的存在是对铜缺乏的一种防御，也是为了保护细胞免受异常高浓度的铜的影响。高尔基体分泌囊泡含有铜ATPase膜蛋白，可能作为细胞内储存铜的隔室。荧光探针可以穿透细胞膜，是研究细胞内金属离子分布的有力工具，但完全缺乏灵敏的在体测量细胞内铜水平的严格分析技术。拟议项目的目标是开发一种铜特定的荧光探针，它不仅可以监测囊泡中不稳定的铜浓度，而且还可以提供有关铜代谢紊乱的细胞系中亚细胞分布的信息。在更大的背景下，铜特异的荧光探针将对长期开发与铜相关的人类疾病的新诊断和治疗工具具有重要意义。