Jia Ben, Jia Lan, Zhu Jingxin
College of Material Science and Engineering, Taiyuan University of Technology, Taiyuan, 030024, P. R. China.
J Fluoresc. 2022 Jan;32(1):389-395. doi: 10.1007/s10895-021-02841-x. Epub 2021 Dec 2.
In this work, the potential application of the fluorescence dye Thioflavin-T (ThT), which can specifically bind to amyloid, as a powerful tool for monitoring secondary structural transitions of silk fibroin (SF) induced by pH in low solution concentrations was examined. Results showed that ThT emission intensities substantially increased when pH decreased from 6.8 to 4.8. This increase may be ascribed to conformational transitions from random coil to β-sheet. The morphology and secondary structure of SF were also investigated via TEM, AFM and circular dichroism spectroscopy. The information obtained herein can be utilized not only for the development of convenient and efficient noninvasive method for monitoring the assembly behavior of SF in aqueous solution but also for in vitro fluorescence imaging.
在这项工作中，研究了荧光染料硫黄素-T（ThT）作为一种强大工具的潜在应用，该染料可特异性结合淀粉样蛋白，用于监测低溶液浓度下pH诱导的丝素蛋白（SF）二级结构转变。结果表明，当pH从6.8降至4.8时，ThT发射强度大幅增加。这种增加可能归因于从无规卷曲到β-折叠的构象转变。还通过透射电子显微镜（TEM）、原子力显微镜（AFM）和圆二色光谱对SF的形态和二级结构进行了研究。本文获得的信息不仅可用于开发方便、高效的非侵入性方法来监测SF在水溶液中的组装行为，还可用于体外荧光成像。
