Development of protein molecular probes for detection of alpha synuclein oligomeric assemblies

 描述(申请人提供)：总体研究目标是开发一种新的分子探针，用于快速、定量、灵敏和特异地检测有毒寡聚形式 阿尔法突触核蛋白(αS)，与帕金森氏症有关。快速、定量、灵敏和特异的αS低聚物检测是首选的，因为(1)准确探测结构不稳定的αS低聚物，(2)实施用于制药应用的高通量分析，以及(3)正确地描绘αS低聚物状态。目前可用的化合物或方法要么不能灵敏地区分具有不同聚集状态的αS形式，要么是不适当的 由于需要额外的样品制备和培养步骤，因此可用于快速、定量检测。由于有毒αS低聚物分子结构的不确定性，要想出一个简单的设计原则来创建高度特异和灵敏的探针是极其困难的。相反，为了开发具有所需性能的新型探针，迫切需要对探针原型的物理化学性质进行详尽的变化，并对这种变化进行快速和可靠的评估。在提交的项目中，PI建议构建一个新的分子平台，人们可以在其中合成和测试大量探针变体，这些变体显示了与αS低聚物结合和后续产生的光学信号相关的广泛物理化学性质。在特定的目标中，将创建和快速测试一组相关的探针结构，以确定在与αS的寡聚形式结合时产生特定光学信号的探针。我们的长期目标是开发一组能够检测不同形式的αS的探针。这样一组探针将能够对治疗剂进行快速可靠的测试，进行早期诊断，并更好地了解αS的寡聚过程。

 DESCRIPTION (provided by applicant): The overall research objective is to develop a novel molecular probe for rapid, quantitative, sensitive and specific detection of toxic oligomeric forms of alpha synuclein (αS), implicated in Parkinson's disease. Rapid, quantitative, sensitive and specific detection of αS oligomers is preferred for (1) accurate probing of structurally unstable αS oligomers, (2) implementation of high-throughput assays for pharmaceutical applications and (3) correct profiling of αS oligomeric states. The currently available compounds or methods either do not sensitively distinguish αS forms in distinct aggregation states or are inappropriate for rapid, quantitative detection due to the requirement of additional sample preparation and incubation steps. Due to the uncertainty associated with the molecular structures of toxic αS oligomers, it is extremely difficult to come up with a simple design principle leading to the creation of highly specific and sensitive probes. Instead, an exhaustive variation of the physico-chemical nature of a probe prototype, and rapid and reliable evaluation of such variation are strongly required to develop novel probes with desired properties. In the submitted project, the PI proposes to construct a new molecular platform where one can synthesize and test a large array of probe variants displaying a wide range of physico-chemical natures associated with binding to αS oligomers and subsequent generation of optical signals. In specific aims, a pool of related probe structures will be created and rapidly tested to identify probes that generate specific optical signals upon binding to oligomeric forms of αS. Our long term goal is to develop a pool of probes capable of detecting distinct forms of αS. Such a panel of probes will allow for rapid and reliable testing of therapeutic agents, developing early diagnostics, and a better understanding of αS oligomerization processes.