Protein diffusion in mammalian cell cytoplasm.
Protein diffusion in mammalian cell cytoplasm.
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DOI：
10.1371/journal.pone.0022962
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发表时间：
2011
期刊：
影响因子：
3.7
通讯作者：
Timonen J
中科院分区：
文献类型：
--
作者：
Kühn T;Ihalainen TO;Hyväluoma J;Dross N;Willman SF;Langowski J;Vihinen-Ranta M;Timonen J
我们引入了一种对整个细胞中蛋白质扩散进行介观建模的新方法。该方法基于根据共焦显微镜数据构建三维数字模型单元。模型细胞被分为细胞质、细胞核、质膜和核膜，其中环境蛋白质运动通过完全数值介观方法进行建模。成像技术无法解析的更精细的细胞结构会显着影响蛋白质运动，在此方法中通过为细胞分配有效的、位置依赖性的孔隙率来解释这种结构。该孔隙率也可以通过共聚焦显微镜使用非结合荧光蛋白的平衡分布来确定。现在可以在该方法中区分细胞液相（细胞溶胶/核溶胶）和细胞质/核质中的扩散。在这里，我们应用该方法来分析光漂白（FRAP）实验后的荧光恢复，其中确定了两种不同细胞系的自由扩散模型蛋白质的扩散系数，并解释了传统 FRAP 结果与荧光相关光谱（FCS）结果之间通常观察到的明显差异。在 FRAP 实验中发现细胞质/核质中的扩散和细胞质/核仁中的扩散之间存在很大差异，所有这些都确定了扩散系数。发现细胞质结果与 FCS 的结果非常一致。
We introduce a new method for mesoscopic modeling of protein diffusion in an entire cell. This method is based on the construction of a three-dimensional digital model cell from confocal microscopy data. The model cell is segmented into the cytoplasm, nucleus, plasma membrane, and nuclear envelope, in which environment protein motion is modeled by fully numerical mesoscopic methods. Finer cellular structures that cannot be resolved with the imaging technique, which significantly affect protein motion, are accounted for in this method by assigning an effective, position-dependent porosity to the cell. This porosity can also be determined by confocal microscopy using the equilibrium distribution of a non-binding fluorescent protein. Distinction can now be made within this method between diffusion in the liquid phase of the cell (cytosol/nucleosol) and the cytoplasm/nucleoplasm. Here we applied the method to analyze fluorescence recovery after photobleach (FRAP) experiments in which the diffusion coefficient of a freely-diffusing model protein was determined for two different cell lines, and to explain the clear difference typically observed between conventional FRAP results and those of fluorescence correlation spectroscopy (FCS). A large difference was found in the FRAP experiments between diffusion in the cytoplasm/nucleoplasm and in the cytosol/nucleosol, for all of which the diffusion coefficients were determined. The cytosol results were found to be in very good agreement with those by FCS.
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影响因子：
3.7
作者：
Dross N;Spriet C;Zwerger M;Müller G;Waldeck W;Langowski J
通讯作者：
Langowski J
DOI：
10.1073/pnas.1006731107
发表时间：
2010-09-21
影响因子：
11.1
作者：
Hinde, Elizabeth;Cardarelli, Francesco;Gratton, Enrico
通讯作者：
Gratton, Enrico
DOI：
10.1073/pnas.051631998
发表时间：
2001-02-27
影响因子：
11.1
作者：
Marsh, BJ;Mastronarde, DN;McIntosh, JR
通讯作者：
McIntosh, JR
影响因子：
64.8
作者：
Phair, RD;Misteli, T
通讯作者：
Misteli, T
影响因子：
3.4
作者：
Beaudouin, JL;Mora-Bermúdez, F;Ellenberg, J
通讯作者：
Ellenberg, J