Decoding the pattern of photon colors in single-molecule FRET.

Decoding the pattern of photon colors in single-molecule FRET.
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DOI:
10.1021/jp903671p
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发表时间:
2009-08-06
期刊:
The journal of physical chemistry. B
影响因子:
--
通讯作者:
Szabo A
Szabo A
中科院分区:
其他
文献类型:
--
作者:
Gopich IV;Szabo A

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单个分子的构象动力学可以用Förster共振能量转移(FRET)来研究,方法是记录分子上的给体和受体染料发出的一系列光子。我们描述了一种简单而稳健的方法来估计不同构象状态之间的跃迁速率以及与这些状态相关的FRET效率。对于具有测量的光子间时间的光子轨迹,通过最大化适当的似然函数来解码颜色图案。这种方法可以用来分析扩散分子产生的光子爆发,以及固定分子产生的光子轨迹。使用对应于两态和三态分子的模拟光子轨迹来说明该过程。即使在由于高背景噪声、构象的光物理性质相似、构象和光子计数率可比而使光子颜色看起来被扰乱的情况下,该方法仍然有效。模型与数据的一致性可以通过对光子轨迹重新着色并将预测的FRET效率直方图与观测的FRET效率直方图进行比较来检查。
Conformational dynamics of a single molecule can be studied using Förster resonance energy transfer (FRET) by recording a sequence of photons emitted by a donor and an acceptor dye attached to the molecule. We describe a simple and robust method to estimate the rates of transitions between different conformational states and the FRET efficiencies associated with these states. For a photon trajectory with measured interphoton times, the pattern of colors is decoded by maximizing the appropriate likelihood function. This approach can be used to analyze bursts of photons from diffusing molecules as well as photon trajectories generated by immobilized molecules. The procedure is illustrated using simulated photon trajectories corresponding to two-state and three-state molecules. The method works even when the photon colors appear to be scrambled because of high background noise, the photophysical properties of the conformers are similar, or the conformational and photon count rates are comparable. The consistency of the model with the data can be checked by recoloring the photon trajectories and comparing the predicted and observed FRET efficiency histograms.
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