Fluorescent N-arylaminonaphthalene sulfonate probes for amyloid aggregation of α-synuclein
Fluorescent N-arylaminonaphthalene sulfonate probes for amyloid aggregation of α-synuclein
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DOI：
10.1529/biophysj.107.125211
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发表时间：
2008-06-15
影响因子：
3.4
通讯作者：
Jovin, Thomas M.
中科院分区：
文献类型：
--
作者：
Celej, M. Soledad;Jares-Erijman, Elizabeth A.;Jovin, Thomas M.
纤维状结构（淀粉样蛋白）的沉积是包括阿尔茨海默病和帕金森病在内的病理状况的特征。蛋白质沉积物的检测及其聚集动力学的评价通常基于荧光探针如硫磺素T和刚果红。在寻找用于研究淀粉样蛋白形成的改进的荧光工具中，我们探索了N-芳基氨基萘磺酸盐（NAS）衍生物作为α-突触核蛋白（AS）纤维化的非共价探针的能力，该纤维化是与帕金森病和其他神经退行性疾病相关的过程。这些化合物以微摩尔K（d）s与纤维状AS结合，并表现出荧光增强、增色和高各向异性。我们的结论是，探针经历的疏水环境和/或限制运动在极性区域。时间和光谱分辨的发射强度和各向异性提供了进一步的信息，关于蛋白质的结构特征和溶剂松弛的动力学。稳态和时间分辨参数在聚集过程中发生变化。与硫磺素T相比，NAS衍生物构成更敏感和通用的AS聚集探针，并在双NAS的情况下检测寡聚体以及纤维状物种。它们可以在方便的连续测定中发挥作用，从而为研究淀粉样蛋白形成的机制和高通量筛选抑制和/或逆转神经退行性疾病中蛋白质聚集的因子提供有用的工具。
The deposition of fibrillar structures (amyloids) is characteristic of pathological conditions including Alzheimer's and Parkinson's diseases. The detection of protein deposits and the evaluation of their kinetics of aggregation are generally based on fluorescent probes such as thioflavin T and Congo red. In a search for improved fluorescence tools for studying amyloid formation, we explored the ability of N-arylaminonaphthalene sulfonate (NAS) derivatives to act as noncovalent probes of a-synuclein (AS) fibrillation, a process linked to Parkinson's disease and other neurodegenerative disorders. The compounds bound to fibrillar AS with micromolar K(d)s, and exhibited fluorescence enhancement, hyperchromism, and high anisotropy. We conclude that the probes experience a hydrophobic environment and/or restricted motion in a polar region. Time- and spectrally resolved emission intensity and anisotropy provided further information regarding structural features of the protein and the dynamics of solvent relaxation. The steady-state and time-resolved parameters changed during the course of aggregation. Compared with thioflavin T, NAS derivatives constitute more sensitive and versatile probes for AS aggregation, and in the case of bis-NAS detect oligomeric as well as fibrillar species. They can function in convenient, continuous assays, thereby providing useful tools for studying the mechanisms of amyloid formation and for high-throughput screening of factors inhibiting and/or reversing protein aggregation in neurodegenerative diseases.