Department of Chemistry, Indian Institute of Science Education and Research Bhopal, Bhopal Bypass Road, Bhauri 462066, MP India.
Department of Biosciences and Bioengineering, Indian Institute of Technology Bombay, Powai, Mumbai 400076, India.
Anal Chem. 2020 Aug 4;92(15):10336-10341. doi: 10.1021/acs.analchem.0c00379. Epub 2020 Jul 22.
The misfolding and aggregation of proteins leading to amyloid formation has been linked to numerous diseases, necessitating the development of tools to monitor the fibrillation process. Here, we report an intramolecular charge transfer (ICT) dye, DMNDC, as an alternative to thioflavin-T (ThT), most commonly used for monitoring amyloid fibrils. Using insulin as a model protein, we show that DMNDC efficiently reports on the kinetics of fibril formation. An approximately 70 nm hypsochromic shift along with a large enhancement in emission intensity was observed upon binding of DMNDC to protein fibrils. The aggregation kinetics of insulin were not significantly affected in the presence of DMNDC, suggesting that DMNDC does not inhibit insulin aggregation. Additionally, the efficient cellular internalization and low toxicity of DMNDC make it highly suited for sensing and imaging of amyloid fibrils in the complex biological milieu.
蛋白质错误折叠和聚集导致淀粉样形成与许多疾病有关，因此需要开发监测纤维形成过程的工具。在这里，我们报告了一种分子内电荷转移（ICT）染料 DMNDC，它可以替代最常用于监测淀粉样纤维的硫黄素 T（ThT）。我们使用胰岛素作为模型蛋白，表明 DMNDC 可以有效地报告纤维形成的动力学。当 DMNDC 与蛋白质纤维结合时，观察到大约 70nm 的蓝移以及发射强度的大幅增强。在 DMNDC 的存在下，胰岛素的聚集动力学没有受到显著影响，这表明 DMNDC 不会抑制胰岛素聚集。此外，DMNDC 具有高效的细胞内化和低毒性，非常适合在复杂的生物环境中感应和成像淀粉样纤维。
