PDF(Pubmed)
Abstract：
Thioflavin T (ThT) informed microviscosity changes can be used to monitor protein aggregation. Steady-state, time-resolved and lasing spectroscopy were used to detect transient states in α-synuclein - a protein associated with Parkinson\'s disease. The major focus was on the nucleation phase, where conventional ThT fluorescence assay lacks appropriate sensitivity to detect early stage oligomers. Instead, lasing spectroscopy and lasing threshold parameters, in particular, were sensitive to detecting protein oligomers. Through lasing spectroscopy, a change in microviscosity correlating with the stages of protein aggregation was observed at two wavelengths 405 and 440 nm. The two wavelengths are associated with free dye molecules and β-sheet bound ThT molecules. This provides a perspective on elucidating the early formed protein aggregation, a critical aspect in understanding the pathogenesis of neurodegenerative diseases. The insights from the presented study shows the potential of using lasing spectroscopy as a sensitive tool in studying protein aggregation dynamics.
摘要：
硫黄素T(ThT)告知微粘度变化可用于监测蛋白质聚集。稳态,时间分辨和激光光谱用于检测α-突触核蛋白的瞬时状态-一种与帕金森病相关的蛋白质。主要关注的是成核阶段，其中常规ThT荧光测定缺乏检测早期寡聚体的适当灵敏度。相反，激光光谱和激光阈值参数，特别是,对检测蛋白质寡聚体敏感。通过激光光谱，在405和440nm两个波长处观察到与蛋白质聚集阶段相关的微粘度变化。这两个波长与游离染料分子和β-折叠结合的ThT分子相关。这为阐明早期形成的蛋白质聚集提供了一个视角，理解神经退行性疾病发病机制的一个关键方面。所提出的研究的见解表明，使用激光光谱法作为研究蛋白质聚集动力学的敏感工具的潜力。
