arXivDaily arXiv每日学术速递 周一至周五更新
arXiv 2107.14770physics.chem-phphysics.bio-phphysics.data-anq-bio.BMq-bio.QM
收藏

在单分子FRET实验中揭示多状态分子动力学-第一部分：FRET线理论

Unraveling multi-state molecular dynamics in single-molecule FRET experiments- Part I: Theory of FRET-Lines

Anders Barth, Oleg Opanasyuk, Thomas-Otavio Peulen, Suren Felekyan, Stanislav Kalinin, Hugo Sanabria, Claus A. M. Seidel

备注：
Main Text (47 pages, 14 figures, 3 tables) with Supporting Information (22 pages, 2 figures)
阅读 PDF
arXiv 页面 ↗HTML 原文 ↗

摘要：

生物分子的构象动力学对它们的功能具有根本的重要性。单分子Förster共振能量转移（smFRET）是一种强大的方法来告知标记分子的结构和动力学。如果动力学发生在亚毫秒的时间尺度上，捕获和量化构象动力学可能是具有挑战性的基于强度的smFRET。多参数荧光检测（MFD）通过同时记录强度和荧光寿命来解决这一挑战。平均供体荧光寿命和荧光强度共同决定了方差，而平均FRET效率则决定了构象动力学。在这里，我们提出了一个一般的框架，涉及平均荧光寿命和强度smFRET计数直方图。使用这个框架，我们展示了如何计算这些观测值的参数关系（FRET线），便于图形化解释的实验数据，可用于测试模型，识别构象状态，解决状态的连接，并可应用于非结构化系统，以推断聚合物链的属性或研究快速蛋白质折叠。为了简化复杂的动力学网络的图形分析，我们推导出一个基于矩的表示的实验数据，并显示如何解耦的运动的荧光标记的构象动力学的生物分子。

英文摘要：

Conformational dynamics of biomolecules are of fundamental importance for their function. Single-molecule Förster Resonance Energy Transfer (smFRET) is a powerful approach to inform on the structure and the dynamics of labeled molecules. If the dynamics occur on the sub-millisecond timescale, capturing and quantifying conformational dynamics can be challenging by intensity-based smFRET. Multiparameter fluorescence detection (MFD) addresses this challenge by simultaneously registering intensities and fluorescence lifetimes. Together, the mean donor fluorescence lifetime and the fluorescence intensities inform on the variance, and the mean FRET efficiency tells the conformational dynamics. Here, we present a general framework that relates average fluorescence lifetimes and intensities in smFRET counting histograms. Using this framework, we show how to compute parametric relations (FRET-lines) of these observables that facilitate a graphical interpretation of experimental data, can be used to test models, identify conformational states, resolve the connectivity of states, and can be applied to unstructured systems to infer properties of polymer chains or study fast protein folding. To simplify the graphical analysis of complex kinetic networks, we derive a moment-based representation of the experimental data and show how to decouple the motion of the fluorescence labels from the conformational dynamics of the biomolecule.

机构、总结、中文摘要均由AI分析而来，可能存在错误，请以论文正文为准。

↑