Organic & Medicinal Chemistry Division , Indian Institute of Chemical Biology (CSIR-IICB) , Kolkata 700032 , India.
Structural Biology and Bioinformatics Division , Indian Institute of Chemical Biology (CSIR-IICB) , 4 Raja S. C. Mullick Road , Kolkata 700032 , India.
Biochemistry. 2019 Feb 26;58(8):1109-1119. doi: 10.1021/acs.biochem.8b01161. Epub 2019 Feb 6.
In this study, we have designed and synthesized a new hybrid ligand (SCG) that can selectively detect cysteine in the free and protein-bound states within minutes at the subnanomolar level. Photoinduced electron transfer was responsible for the visible color change as well as a large increase in steady state fluorescence. This detection was validated by using multiple model protein systems with differing cysteine environments and spatial arrangements. SCG was able to monitor the early events of the folding/aggregation kinetics of α-synuclein, a protein involved in the pathology of Parkinson's disease. The early events consisted of conformational fluctuations between different forms of the protein and oligomer formation. SCG was found to be effective in detecting early isomers of α-syn in vitro and in live cell environments.
在这项研究中,我们设计并合成了一种新的混合配体(SCG),它可以在亚纳摩尔水平下在数分钟内选择性地检测游离态和蛋白质结合态的半胱氨酸。光诱导电子转移负责可见颜色变化以及稳态荧光的大幅增加。通过使用具有不同半胱氨酸环境和空间排列的多种模型蛋白系统,验证了这种检测。SCG 能够监测涉及帕金森病病理学的α-突触核蛋白的折叠/聚集动力学的早期事件。早期事件包括不同形式的蛋白质之间的构象波动和寡聚体形成。发现 SCG 可有效检测体外和活细胞环境中α-突触核蛋白的早期异构体。