Diffusion analysis of glucocorticoid receptor and antagonist effect in living cell nucleus
Diffusion analysis of glucocorticoid receptor and antagonist effect in living cell nucleus
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DOI：
10.1016/j.yexmp.2006.12.001
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发表时间：
2007-04-01
影响因子：
3.6
通讯作者：
Kinjo, Masataka
中科院分区：
文献类型：
--
作者：
Mikuni, Shintaro;Pack, Changi;Kinjo, Masataka
分析了hGR α在活细胞中的扩散特性。加入Dex后，hGR α与RU 486一样从胞浆转移到细胞核;但细胞核中蛋白质的布朗运动有所不同。为了分析活细胞核内的微环境，构建了四种不同的串联EGFPs。串联EGFP的扩散依赖于蛋白质在溶液中作为棒状分子的长度。我们发现两种流动性，快速扩散流动性作为一个主要组成部分，慢得多的扩散流动性作为一个主要组成部分，在活细胞核质。在此基础上，我们通过扩散常数的分布比较了在Dex或RU 486存在下hGR α在细胞核中的扩散性质。提示RU 486可激活EGFP-hGR α，并使其保持在结合辅因子、GRE和最终复合物的阶段。最后，这意味着二聚化不是与GRE缔合所必需的，尽管它是EGFP-hGR α复合物稳定所必需的。(C)2006年爱思唯尔公司All rights reserved.
The diffusion properties of hGR alpha in living cells have been analyzed. The hGR alpha translocalized from the cytosol to the nucleus after addition of Dex just as RU486; however, the Brownian motions of the proteins in nucleus were different. In order to analysis microenvironment of the nucleus of living cell, four different tandem EGFPs were constructed. Diffusion of tandem EGFP was dependent on the length of the protein as a rod-like molecule in solution. We found two kinds of mobility, fast diffusional mobility as a major component and much slower diffusional mobility as a major component in living cells nucleoplasm. On the bases of this analysis, we compared the diffusion property of hGR alpha in the nucleus at the presence of Dex or RU486 by distribution of diffusion constants. Our result may suggest that EGFP-hGR alpha is activated by RU486 and kept the stage of binding cofactor, GRE and final complex. Finally this means that dimerization is not required for association with GRE, although it is required for stabilization of a complex of EGFP-hGR alpha. (C) 2006 Elsevier Inc. All rights reserved.