Fluorescence Lifetime of Fluorescent Proteins as an Intracellular Environment Probe Sensing the Cell Cycle Progression
Fluorescence Lifetime of Fluorescent Proteins as an Intracellular Environment Probe Sensing the Cell Cycle Progression
复制标题
DOI：
10.1021/cb300065w
复制
发表时间：
2012-08-01
影响因子：
4
通讯作者：
Prasad, Paras N.
中科院分区：
文献类型：
--
作者：
Pliss, Artem;Zhao, Lingling;Prasad, Paras N.
荧光蛋白的荧光寿命受介质中溶质浓度的影响，与局部折射率成反比。在本文中，我们介绍了使用这种依赖性来探测细胞分子环境及其在细胞过程中的转换的概念。我们利用培养细胞中表达的绿色荧光蛋白和tdTomato荧光蛋白的荧光寿命，并探测细胞周期进程中局部分子环境的变化。我们报告说，最长的荧光寿命发生在有丝分裂。随着细胞分裂，这些蛋白质的荧光寿命迅速缩短。tdTomato在核质中的荧光寿命在整个S期内逐渐增加，并一直持续到间期结束。我们解释所观察到的荧光寿命的变化来自整个细胞周期进程中的细胞内部的大分子溶质的浓度变化。
The fluorescence lifetime of fluorescent proteins is affected by the concentration of solutes in a medium, in inverse correlation with local refractive index. In this paper, we introduce the concept of using this dependence to probe cellular molecular environment and its transformation during cellular processes. We employ the fluorescence lifetime of Green Fluorescent Protein and tdTomato Fluorescent Protein expressed in cultured cells and probe the changes in the local molecular environment during the cell cycle progression. We report that the longest fluorescence lifetimes occurred during mitosis. Following the cell division, the fluorescence lifetimes of these proteins were rapidly shortened. Furthermore the fluorescence lifetime of tdTomato in the nucleoplasm gradually increased throughout the span of S-phase and remained constantly long until the end of interphase. We interpret the observed fluorescence lifetime changes to be derived from changes in concentration of macromolecular solutes in the cell interior throughout cell cycle progression.