Tools for the Detection of Ethylene

7.项目总结/摘要 虽然乙烯长期以来一直被认为是一种重要的植物激素， 在哺乳动物体内产生的氧化应激是许多疾病的标志。在 特别的乙烯是由脂质过氧化物和/或它们的中间体的自由基断裂产生的。 阵脂质过氧化物的形成是活性氧物质的结果，其涉及 在许多疾病中起着压力或信号作用，包括癌症，心血管疾病， 神经退行性疾病等。虽然有一些复杂的光谱方法 为了灵敏地测量呼出气中的生物标志物乙烯，这些方法是必要的。 受空间分辨率和样本复杂度的限制。最近，我们的团队开发了一个 荧光化学剂量计能够检测活细胞中的乙烯;然而， 检测内源性乙烯的灵敏度不足。 因此，本探索性研究提案的总体目标是确定结构和 靶向修饰可以提供必要的灵敏度，以研究内源性 乙烯和任何潜在的信号作用。为了实现这一目标，我们的目标是（1）结构优化 配体的钌和调整附加的荧光团的电子物理性质;和（2） 为了使用探针的亚细胞靶向至预期乙烯存在于细胞中的细胞结构域， 更高的浓度。 预计所提出的研究将导致探针具有显著改善的限制。 检测（~2个数量级）的范围内，可能是预期的内源性乙烯水平， 疾病状态。这将主要通过合成修饰的探针来实现， 所得性质的表征和活细胞中的定位研究。这项探索性研究是 本研究的目的是评估内源乙烯检测的可行性，为进一步研究内源乙烯的含量奠定基础。 进一步研究乙烯在细胞水平上的产生。检测乙烯的能力 使用当前方法无法获得的空间分辨率将提供一种广泛适用的工具， 报告脂质过氧化反应。 1

7. Project Summary/Abstract While ethylene has long been known as an important plant hormone it has also been demonstrated to be produced in mammals as a result of oxidative stress that is hallmark to numerous diseases. In particular ethylene arises from the radical fragmentation of lipid peroxides and/or intermediates in their formation. The formation of lipid peroxides is a result of reactive oxygen species, which are implicated as playing stress or signaling roles in numerous diseases including cancer, cardiovascular disease, and neurodegenerative diseases amongst others. While there are some sophisticated spectroscopic methods for sensitively measuring the biomarker ethylene in exhaled breath, these approaches are necessarily limited in spatial resolution and complexity of sample. Recently, our group has developed a profluorescent chemodosimeter that is capable of detecting ethylene in live cells; however the current sensitivity is insufficient for detection of endogenous ethylene. Therefore the overall goal of this exploratory research proposal is to determine if structural and targeting modifications can provide the necessary sensitivity to study the endogenous production of ethylene and any potential signaling roles. To achieve this goal we aim to (1) structurally optimize the ligands about the ruthenium and tune the photophysical properties of the appended fluorophore; and (2) to use subcellular targeting of the probe to cellular domains where ethylene is expected to be present in higher concentrations. It is expected that the proposed research will result in probes with significantly improved limit of detection (~2 orders of magnitude) in the range that might be expected for endogenous ethylene levels in disease states. This will primarily be accomplished through synthesis of modified probes, characterization of resulting properties, and localization studies in live cells. This exploratory research is expected to evaluate the feasibility of detecting endogenous ethylene levels and lay the groundwork for further investigations into ethylene's production at the cellular level. The ability to detect ethylene with spatial resolution not available using current approaches would provide a broadly applicable tool capable of reporting lipid peroxidation. 1