Real-Time Monitoring of Alzheimer's-Related Amyloid Aggregation via Probe Enhancement-Fluorescence Correlation Spectroscopy.
Real-Time Monitoring of Alzheimer's-Related Amyloid Aggregation via Probe Enhancement-Fluorescence Correlation Spectroscopy.
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DOI：
10.1021/acschemneuro.5b00176
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发表时间：
2015-09-16
影响因子：
5
通讯作者：
Shah JV
中科院分区：
文献类型：
--
作者：
Guan Y;Cao KJ;Cantlon A;Elbel K;Theodorakis EA;Walsh DM;Yang J;Shah JV
本工作描述了使用荧光相关光谱（FCS）和一种新的淀粉样蛋白结合荧光探针ARCAM 1来监测阿尔茨海默病相关淀粉样蛋白β肽（Aβ）的聚集。ARCAM 1在与Aβ组装体结合时表现出荧光发射的大幅增加，使其成为探针增强FCS（PE-FCS）的优秀候选物。ARCAM 1结合不改变Aβ聚集动力学。当与硫磺素T（ThT）或用荧光团修饰的Aβ肽相比时，它还表现出更大的动态范围作为通过FCS报告Aβ中聚集体大小的探针。使用荧光猝发分析（通过PE-FCS）跟踪Aβ的聚集，与典型的整体荧光测量相比，我们在显著更早的时间点检测到可溶性聚集体。自相关分析揭示了这些早期Aβ组装体的大小。这些结果表明，基于PE-FCS/ARCAM 1的测定可以检测组装过程中小Aβ聚集中间体并提供其尺寸表征，这可以监测和研究在阿尔茨海默病和其他神经退行性疾病患者的生物流体中瞬时积累的这种聚集体。
This work describes the use of fluorescence correlation spectroscopy (FCS) and a novel amyloid-binding fluorescent probe, ARCAM 1, to monitor the aggregation of the Alzheimer's disease-associated amyloid β-peptide (Aβ). ARCAM 1 exhibits a large increase in fluorescence emission upon binding to Aβ assemblies, making it an excellent candidate for probe enhancement FCS (PE-FCS). ARCAM 1 binding does not change Aβ aggregation kinetics. It also exhibits greater dynamic range as a probe in reporting aggregate size by FCS in Aβ, when compared to thioflavin T (ThT) or an Aβ peptide modified with a fluorophore. Using fluorescent burst analysis (via PE-FCS) to follow aggregation of Aβ, we detected soluble aggregates at significantly earlier time points compared to typical bulk fluorescence measurements. Autocorrelation analysis revealed the size of these early Aβ assemblies. These results indicate that PE-FCS/ARCAM 1 based assays can detect and provide size characterization of small Aβ aggregation intermediates during the assembly process, which could enable monitoring and study of such aggregates that transiently accumulate in biofluids of patients with Alzheimer's and other neurodegenerative diseases.