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设计和合成一种FlAsH型Mg2+光探针,用于标记特定蛋白质

Tomohiko Fujii¹, Yutaka Shindo, Kohji Hotta

¹Department of Biosciences and Informatics, §Department of Applied Chemistry, and ∥Department of Chemistry, Faculty of Science and Technology Keio University , 3-14-1 Hiyoshi, Kohoku-ku, Yokohama 223-8522, Japan.

Journal of the American Chemical Society

|January 23, 2014

概括

研究人员开发了一种新的光探针KMG-104-AsH,以可视化细胞内的离子 (Mg2+) 变化. 这个工具专门针对蛋白质,提供了Mg2+动态和细胞过程的见解.

科学领域:

细胞生物学细胞生物学
生物化学生物化学
分子成像学分子成像学

背景情况:

离子 (Mg2+) 对于细胞功能至关重要,但其细胞内动力学仍然不太清楚.
现有的方法缺乏Mg2+调动的具体性和实时跟踪能力.

研究的目的:

开发和验证一种新的光探针,用于细胞内Mg2+度的分子成像.
为了研究活细胞内Mg2+的动态和定位.

主要方法:

合成和表征KMG-104-AsH探针,将Mg2+选择性光体与四半胺标签 (TCtag) 结合部分结合在一起.
在溶液中的光学特性分析和在HeLa细胞中使用TCtag融合蛋白进行活细胞成像的应用.
监测由FCCP诱导的线粒体中的Mg2+释放.

主要成果:

在结合Mg2+和TCtag时,KMG-104-AsH的光度增加了10倍以上,对Mg2+比Ca2+具有很高的选择性.
探针具有膜透性,并且特别与细胞质和线粒体中的TCtag标记蛋白质结合.
图像检测显示,Mg2+从线粒体释放出来,作为FCCP治疗的反应.

结论:

KMG-104-AsH是一个强大的工具,用于实时,特定地点的细胞内Mg2+成像.
探测器有助于阐明Mg2+的动态和调节机制在各种细胞区.
这项技术为研究依赖Mg2+的生理过程开辟了新的途径.

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