Quantitative FRET studies and integrative modeling unravel the structure and dynamics of biomolecular systems
Quantitative FRET studies and integrative modeling unravel the structure and dynamics of biomolecular systems
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DOI：
10.1016/j.sbi.2016.11.012
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发表时间：
2016-10-01
影响因子：
6.8
通讯作者：
Seidel, Claus A. M.
中科院分区：
文献类型：
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作者：
Dimura, Mykola;Peulen, Thomas O.;Seidel, Claus A. M.
福斯特共振能量转移 (FRET) 与单分子光谱相结合，可探测大分子结构和动力学，并识别共存的构象状态。我们通过满足 FRET 对网络（混合 FRET）的空间限制来回顾集成结构建模的最新方法发展。我们讨论结合先验结构知识并获得最佳距离网络的程序。最后，提出了混合 FRET 的工作流程，可自动执行集成结构建模和实验规划，以将混合 FRET 置于轨道上。为了测试这个工作流程，我们模拟了真实的单分子实验并解析了三种蛋白质构象异构体，以 30 μs 和 10 ms 的速度进行交换，与目标结构相比，RMSD 的准确度为 1-3 埃。还讨论了其他光谱和成像数据的合并。
Forster Resonance Energy Transfer (FRET) combined with single-molecule spectroscopy probes macromolecular structure and dynamics and identifies coexisting conformational states. We review recent methodological developments in integrative structural modeling by satisfying spatial restraints on networks of FRET pairs (hybrid-FRET). We discuss procedures to incorporate prior structural knowledge and to obtain optimal distance networks. Finally, a workflow for hybrid-FRET is presented that automates integrative structural modeling and experiment planning to put hybrid-FRET on rails. To test this workflow, we simulate realistic single-molecule experiments and resolve three protein conformers, exchanging at 30 mu s and 10 ms, with accuracies of 1-3 angstrom RMSD versus the target structure. Incorporation of data from other spectroscopies and imaging is also discussed.