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Efficient Detection of Early Events of alpha-Synuclein Aggregation Using a Cysteine Specific Hybrid Scaffold

机译:使用半胱氨酸特定杂交支架有效地检测α-突触核蛋白聚集的早期事件

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摘要

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 alpha-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 alpha-syn in vitro and in live cell environments.
机译:在该研究中,我们设计并合成了一种新的杂交配体(SCG),其可以在亚甲醛水平的分钟内选择性地检测自由和蛋白质结合状态的半胱氨酸。 光抑制电子转移负责可见的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布置的多种模型蛋白质系统来验证该检测。 SCG能够监测α-突触核蛋白的折叠/聚集动力学的早期事件,这是帕金森病病理学的蛋白质。 早期事件包括不同形式的蛋白质和低聚物形成之间的构象波动。 发现SCG有效地检测体外和活细胞环境中α-SYN的早期异构体。

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  • 来源
    《Biochemistry》 |2019年第8期|共11页
  • 作者单位

    Indian Inst Chem Biol CSIR IICB Organ &

    Med Chem Div Kolkata 700032 India;

    Indian Inst Chem Biol CSIR IICB Struct Biol &

    Bioinformat Div 4 Raja SC Mullick Rd Kolkata 700032 India;

    Indian Inst Chem Biol CSIR IICB Canc Biol &

    Inflammatory Disorder Div 4 Raja SC Mullick Rd Kolkata 700032 India;

    Indian Inst Chem Biol CSIR IICB Canc Biol &

    Inflammatory Disorder Div 4 Raja SC Mullick Rd Kolkata 700032 India;

    Indian Inst Chem Biol CSIR IICB Organ &

    Med Chem Div Kolkata 700032 India;

    Indian Inst Chem Biol CSIR IICB Struct Biol &

    Bioinformat Div 4 Raja SC Mullick Rd Kolkata 700032 India;

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  • 原文格式 PDF
  • 正文语种 eng
  • 中图分类 生物化学;
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