Characterizing fluorescence recovery curves for nuclear proteins undergoing binding events
Characterizing fluorescence recovery curves for nuclear proteins undergoing binding events
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DOI：
10.1016/j.bulm.2004.02.005
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发表时间：
2004-11-01
影响因子：
3.5
通讯作者：
De Vries, G
中科院分区：
文献类型：
--
作者：
Carrero, G;Crawford, E;De Vries, G
光漂白后荧光恢复(FRAP)是一种用来测量细胞核内蛋白质流动性的实验技术。在对感兴趣的蛋白质进行荧光标记以进行可视化和监测后，对细胞核的一小部分进行光漂白。实验的FRAP数据是通过记录该区域随时间的荧光恢复而获得的。在这篇文章中，我们描述了具有近似空间均匀结构的扩散核蛋白经历结合事件的荧光恢复曲线。我们分析了解释FRAP实验数据的两种数学模型，即反应扩散模型和隔室模型。微扰分析清楚地解释了实验恢复曲线所表现出的两种重要的极限动力学行为类型，即(1)减弱的扩散恢复和(2)以快相和慢相为特征的两相恢复。我们展示了这两个使用不同方法描述同一类型动态的模型如何联系和共享共同点。所得结果可用于从蛋白质动力学的角度解释实验FRAP数据，并简化参数估计任务。这一结果在核肌动蛋白和组蛋白HI中得到了应用。(C)2004年数学生物学学会。爱思唯尔有限公司出版。保留所有权利。
Fluorescence recovery after photobleaching (FRAP) is an experimental technique used to measure the mobility of proteins within the cell nucleus. After proteins of interest are fluorescently tagged for their visualization and monitoring, a small region of the nucleus is photobleached. The experimental FRAP data are obtained by recording the recovery of the fluorescence in this region over time. In this paper, we characterize the fluorescence recovery curves for diffusing nuclear proteins undergoing binding events with an approximate spatially homogeneous structure. We analyze two mathematical models for interpreting the experimental FRAP data, namely a reaction-diffusion model and a compartmental model. Perturbation analysis leads to a clear explanation of two important limiting dynamical types of behavior exhibited by experimental recovery curves, namely, (1) a reduced diffusive recovery, and (2) a biphasic recovery characterized by a fast phase and a slow phase. We show how the two models, describing the same type of dynamics using different approaches, relate and share common ground. The results can be used to interpret experimental FRAP data in terms of protein dynamics and to simplify the task of parameter estimation. Application of the results is demonstrated for nuclear actin and type HI histone. (C) 2004 Society for Mathematical Biology. Published by Elsevier Ltd. All rights reserved.