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Abstract：
Understanding the structure of biomolecules is vital for deciphering their roles in biological systems. Single-molecule techniques have emerged as alternatives to conventional ensemble structure analysis methods for uncovering new biology in molecular dynamics and interaction studies, yet only limited structural information could be obtained experimentally. Here, we address this challenge by introducing iMAX FRET, a one-pot method that allows ab initio 3D profiling of individual molecules using two-color FRET measurements. Through the stochastic exchange of fluorescent weak binders, iMAX FRET simultaneously assesses multiple distances on a biomolecule within a few minutes, which can then be used to reconstruct the coordinates of up to four points in each molecule, allowing structure-based inference. We demonstrate the 3D reconstruction of DNA nanostructures, protein quaternary structures, and conformational changes in proteins. With iMAX FRET, we provide a powerful approach to advance the understanding of biomolecular structure by expanding conventional FRET analysis to three dimensions.
摘要：
了解生物分子的结构对于破译它们在生物系统中的作用至关重要。单分子技术已成为传统集成结构分析方法的替代方法，可在分子动力学和相互作用研究中发现新的生物学。然而，只有有限的结构信息可以通过实验获得。这里,我们通过引入iMAXFRET来应对这一挑战，一种一锅法，允许使用双色FRET测量对单个分子进行从头开始3D分析。通过荧光弱结合剂的随机交换，iMAXFRET在几分钟内同时评估生物分子上的多个距离，然后可以用来重建每个分子中最多四个点的坐标，允许基于结构的推理。我们展示了DNA纳米结构的三维重建，蛋白质四级结构，和蛋白质的构象变化。有了iMAXFRET,通过将常规FRET分析扩展到三维，我们提供了一种强大的方法来促进对生物分子结构的理解。
