ABEL-FRET: tether-free single-molecule FRET with hydrodynamic profiling.
ABEL-FRET: tether-free single-molecule FRET with hydrodynamic profiling.
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ABEL-FRET：具有流体动力学分析的无束缚单分子 FRET。
DOI：
10.1038/s41592-021-01173-9
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发表时间：
2021
期刊：
影响因子：
48
通讯作者：
Wang,Quan
中科院分区：
文献类型：
--
作者：
Wilson,Hugh;Wang,Quan
单分子Förster共振能量转移（smFRET）已成为一种通用和广泛的方法来探测纳米级构象和动力学。然而，目前的实验方式往往诉诸分子固定长时间的观察，并不总是接近基于FRET的纳米尺度计量的分辨率极限。在这里，我们提出了ABEL-FRET，一个无固定化的smFRET测量平台，具有很高的FRET效率分辨能力。重要的是，单分子的扩散率是用来提供额外的大小和形状信息的单个分子的流体动力学分析，这与同时测量的分子内构象，通过FRET，使生物分子及其复合物的整体和动态视图。
Single-molecule Förster resonance energy transfer (smFRET) has become a versatile and widespread method to probe nanoscale conformation and dynamics. However, current experimental modalities often resort to molecule immobilization for long observation times and do not always approach the resolution limit of FRET-based nanoscale metrology. Here we present ABEL-FRET, an immobilization-free platform for smFRET measurements with ultrahigh resolving power in FRET efficiency. Importantly, single-molecule diffusivity is used to provide additional size and shape information for hydrodynamic profiling of individual molecules, which, together with the concurrently measured intramolecular conformation through FRET, enables a holistic and dynamic view of biomolecules and their complexes.