可见光和近红外可激活的子胺探针用于活细胞中的化蛋白标记
1Morningside Laboratory for Chemical Biology, Department of Chemistry, The University of Hong Kong, Pokfulam Road, Hong Kong, China.
Journal of the American Chemical Society
|September 2, 2020
概括
研究人员使用可见光或近红外光开发了新型的化化合物,用于活细胞内源性蛋白质的选择性标记. 这种方法可以在没有紫外线细胞毒性的情况下精确地可视化和量化蛋白质水平和动态.
科学领域:
- 化学生物学
- 分子成像
- 细胞动力学
背景情况:
- 在活细胞中选择性化学标记是具有挑战性的.
- 现有的方法通常依赖于基因工程或紫外线,造成局限性.
研究的目的:
- 开发一种新的生物对等光辐射方法,用于对内源性蛋白质进行选择性和化标记.
- 能够精确地视觉化和量化活细胞中的蛋白质水平和动态.
主要方法:
- 使用可见 (430-490 nm) 和近红外 (800 nm) 光激活的新型二氧化化合物.
- 通过碳中间体的插入开发了化标签策略.
- 将该方法应用于各种细胞类型中的蛋白质 (CA-II,eDHFR).
- 已证明活细胞成像,流细胞计量和动态蛋白调节的检测 (低氧诱导的CA- IX积累).
- 使用双光子激发进行细胞选择性,空间控制的辐射.
主要成果:
- 实现活细胞内源性蛋白质的快速,选择性和性标记.
- 实现了精确的细胞下定位可视化和可靠的蛋白质定量.
- 成功检测出动态蛋白质变化,例如因低氧引起的CA- IX积累.
- 使用近红外光进行空间控制的细胞选择性标记.
结论:
- 开发的生物直角光辐射方法为内源性蛋白质标记提供了强大的工具.
- 这种方法可以规避紫外线细胞毒性,避免需要外部报告者.
- 开辟了化学生物学和分子成像领域的新发现和机械研究的途径.
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