Kinetics of NF-κB nucleocytoplasmic transport probed by single-cell screening without imaging.
Kinetics of NF-κB nucleocytoplasmic transport probed by single-cell screening without imaging.
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DOI：
10.1039/c0lc00094a
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发表时间：
2010-11-07
期刊：
影响因子：
6.1
通讯作者：
Lu C
中科院分区：
文献类型：
--
作者：
Wang J;Fei B;Zhan Y;Geahlen RL;Lu C
蛋白质和RNA在细胞核和细胞质之间的运输(核质运输)对细胞的各种功能是至关重要的。由于缺乏在单个细胞水平上测量细胞群体中细胞内蛋白质的核和胞质部分的定量工具，阻碍了对这一过程中涉及的动力学的研究。在这份报告中，我们描述了一种新的方法，微流控电穿孔流式细胞术，研究了重要的转录因子NF-κB的核质转运动力学。利用从单细胞收集的数据，我们定量地描述了群体平均的动力学参数，如核转录因子-κB转运的速率常数和表观激活势垒。我们的数据表明，NF-κB核质转运符合一级动力学，是一个受平衡热力学支配的相当可逆的过程。
Transport of protein and RNA cargoes between the nucleus and cytoplasm (nucleocytoplasmic transport) is vital for a variety of cellular functions. The studies of kinetics involved in such processes have been hindered by the lack of quantitative tools for measurement of the nuclear and cytosolic fractions of an intracellular protein at the single cell level for a cell population. In this report, we describe using a novel method, microfluidic electroporative flow cytometry, to study kinetics of nucleocytoplasmic transport of an important transcription factor NF-κB. With data collected from single cells, we quantitatively characterize the population-averaged kinetic parameters such as the rate constants and apparent activation barrier for NF-κB transport. Our data demonstrate that NF-κB nucleocytoplasmic transport fits first-order kinetics very well and is a fairly reversible process governed by equilibrium thermodynamics.
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影响因子：
4.4
作者：
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通讯作者：
Fitzgerald-Bocarsly, Patricia
影响因子：
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