通过能量转移测量的全局分析同时测定分子内距离分布和构象动力学。
Simultaneous determination of intramolecular distance distributions and conformational dynamics by global analysis of energy transfer measurements.
作者信息
Beechem J M, Haas E
机构信息
University of Illinois Urbana-Champaign, Department of Physics 61801.
出版信息
Biophys J. 1989 Jun;55(6):1225-36. doi: 10.1016/S0006-3495(89)82918-2.
Abstract
Fluorescence energy transfer is widely used for determination of intramolecular distances in macromolecules. The time dependence of the rate of energy transfer is a function of the donor/acceptor distance distribution and fluctuations between the various conformations which may occur during the lifetime of the excited state. Previous attempts to recover both distance distributions and segmental diffusion from time-resolved experiments have been unsuccessful due to the extreme correlation between fitting parameters. A method has been developed, based on global analysis of both donor and acceptor fluorescence decay curves, which overcomes this extreme cross-correlation and allows the parameters of the equilibrium distance distributions and intramolecular diffusion constants to be recovered with high statistical significance and accuracy. Simulation studies of typical intramolecular energy transfer experiments reveal that both static and dynamic conformational distribution information can thus be obtained at a single temperature and viscosity.
摘要
荧光能量转移被广泛用于测定大分子中的分子内距离。能量转移速率的时间依赖性是供体/受体距离分布以及在激发态寿命期间可能出现的各种构象之间波动的函数。由于拟合参数之间存在极强的相关性，以往从时间分辨实验中恢复距离分布和片段扩散的尝试均未成功。基于对供体和受体荧光衰减曲线的全局分析，已开发出一种方法，该方法克服了这种极强的交叉相关性，并能以高统计显著性和准确性恢复平衡距离分布和分子内扩散常数的参数。对典型分子内能量转移实验的模拟研究表明，在单一温度和粘度下即可获得静态和动态构象分布信息。
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