定量 FRET 研究和综合建模揭示了生物分子系统的结构和动态。
Quantitative FRET studies and integrative modeling unravel the structure and dynamics of biomolecular systems.
机构信息
Chair for Molecular Physical Chemistry, Heinrich Heine University Düsseldorf, 40225 Düsseldorf, Germany; Institute for Pharmaceutical and Medicinal Chemistry, Heinrich Heine University Düsseldorf, 40225 Düsseldorf, Germany.
Chair for Molecular Physical Chemistry, Heinrich Heine University Düsseldorf, 40225 Düsseldorf, Germany.
出版信息
Curr Opin Struct Biol. 2016 Oct;40:163-185. doi: 10.1016/j.sbi.2016.11.012. Epub 2016 Dec 7.
Förster 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 30μs and 10ms, with accuracies of 1-3Å RMSD versus the target structure. Incorporation of data from other spectroscopies and imaging is also discussed.
Förster 共振能量转移（FRET）与单分子光谱学相结合，可探测大分子结构和动态，并确定共存的构象状态。我们综述了通过满足 FRET 对网络（杂交-FRET）的空间约束来进行综合结构建模的最新方法学进展。我们讨论了结合先验结构知识和获得最佳距离网络的程序。最后，提出了一个杂交-FRET 的工作流程，该流程可以实现综合结构建模和实验规划的自动化，从而使杂交-FRET 得以实施。为了测试这个工作流程，我们模拟了现实的单分子实验，并以相对于目标结构的 RMSD 为 1-3Å 的精度，解析了以 30μs 和 10ms 交换的三种蛋白质构象。我们还讨论了结合其他光谱学和成像数据的方法。
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