Effect of Diffusion on Resonance Energy Transfer Rate Distributions: Implications for Distance Measurements.
Effect of Diffusion on Resonance Energy Transfer Rate Distributions: Implications for Distance Measurements.
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DOI：
10.1021/acs.jpcb.5b06567
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发表时间：
2015-10-01
期刊：
影响因子：
--
通讯作者：
Hilser VJ
中科院分区：
文献类型：
--
作者：
Toptygin D;Chin AF;Hilser VJ
Intrinsically disordered protein regions and many other biopolymers lack the three-dimensional structure that could be determined by X-ray crystallography or NMR, which encourages the application of alternative experimental methods. Time-resolved resonance energy transfer data are often used to measure distances between two fluorophores attached to a flexible biopolymer. This is complicated by the rotational and translational diffusion of the fluorophores and by non-monoexponential donor decay in the absence of the acceptor. Equation is derived here, which is applicable regardless of whether ID(t) is monoexponential. ID(t) and IDA(t)are the δ-excitation donor emission decays in the absence and in the presence of the acceptor; F(t) contains information about energy transfer, donor–acceptor distance distribution, and diffusion dynamics. It is shown that in the absence of rotational and translational diffusion, F(t) is a continuous distribution of exponentials, whereas in the presence of rotational and translational diffusion, F(t) is a sum of discrete exponentials. For each case it is shown how F(t) is related to the distance distribution. Experimental data obtained with a flexible tetradecapeptide in aqueous solution clearly demonstrate that F(t) is a sum of discrete exponential terms. A partial differential equation describing resonance energy transfer in the presence of both rotational and translational diffusion of the donor and acceptor tethered to the ends of a semiflexible chain is solved in this work using a combination of analytical and numerical methods; the solution is used to fit time-resolved emission of the donor, which makes it possible to determine the model parameters: contour length, persistence length, and the end-to-end translational diffusion coefficient.
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