尼尔,基因编码的分支链氨基酸指标基于mNeon绿色光蛋白和LIVBP蛋白
Aysilu N Asanova1, Oksana M Subach1, Sofya A Myachina1
1Complex of NBICS Technologies, National Research Center "Kurchatov Institute", Moscow 123182, Russia.
ACS sensors
|October 14, 2024
概括
研究人员开发了NeIle,这是一种用于分支链氨基酸 (BCAA) 的新型光传感器. 这种新工具为可视化细胞中的BCAA提供了更广泛的动态范围,推动了代谢研究.
科学领域:
- 生物化学 生物化学
- 分子生物学分子生物学
- 细胞生物学 细胞生物学
背景情况:
- 分支链氨基酸 (BCAA) 对哺乳动物细胞和中枢神经系统的功能至关重要.
- 现有的基因编码的BCAA指标具有局限性,特别是有限的动态范围.
研究的目的:
- 开发一种新的,基于单一光蛋白的BCAA传感器,具有改进的动态范围.
- 描述传感器的特性,并可视化活细胞中的BCAA动态.
主要方法:
- 尼尔传感器的开发是通过将循环 permuted mNeonGreen 与氨酸-单氨酸-氨酸结合蛋白 (LIVBP) 融合而成的.
- 在溶液和活哺乳动物细胞中对NeIle的生物化学和光谱表征.
- 进行X射线晶体学以确定NeIle在异黄素结合状态中的结构.
主要成果:
- 尼尔表现出对高动态范围 (27,19和11倍) 的异黄素,素和素的阳性光反应.
- 传感器对白氨酸 (2.9 mM) 和异氨酸 (5.0 mM) 呈现出高度亲和力.
- 尼尔成功地可视化了包括神经元在内的活哺乳动物细胞内的各种器官中的异黄素动力学.
结论:
- 尼尔 (NeIle) 是一种对BCAA的敏感和多功能光传感器,克服了以前指标的局限性.
- 传感器的结构导向设计为其响应机制和选择性提供了洞察力.
- 尼尔能够实时监测细胞和亚细胞区内的BCAA动态.
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