近日，美国圣犹达儿童研究医院教授Scott C. Blanchard及其团队的研究开发出了单分子尺度下对多种生物系统的并行停流检测。相关论文于2025年12月2日发表于国际顶尖学术期刊《自然—方法学》杂志上。

为了解决这些限制，研究人员引入了平行快速交换单分子荧光和单分子FRster共振能量转移，使各种系统的稳态和预稳态询问同时进行。利用这种方法，课题组阐明了支持β-arrestin1激活的不同构象事件的时间，揭示了抗生素诱导的对信使RNA解码保真度的影响，并证明内源性编码核糖体RNA序列变异调节抗生素敏感性。这种可推广和可扩展的方法有望扩大生物分子功能定量单分子询问的范围和可重复性。

据悉，单分子成像技术对不同生物系统的组成和构象的功能变化提供了前所未有的见解。与其他生物物理方法一样，单分子荧光和FRster共振能量转移调查通常仅限于一次检查一个样品。因此，实验吞吐量受到限制，并且引入的实验方差可以模糊密切相关系统中的功能差异。

附：英文原文

Title: Parallel stopped-flow interrogation of diverse biological systems at the single-molecule scale

Author: Kiselev, Roman, Brady, Ryan A., Modak, Arnab, Cruz-Navarrete, F. Aaron, Alejo, Jose L., Terry, Daniel S., Altman, Roger B., Asher, Wesley B., Javitch, Jonathan A., Blanchard, Scott C.

Issue&Volume: 2025-12-02

Abstract: Single-molecule imaging techniques have provided unprecedented insights into functional changes in composition and conformation across diverse biological systems. As with other biophysical methods, single-molecule fluorescence and Frster resonance energy transfer investigations are typically limited to examination of one sample at a time. Consequently, experimental throughput is restricted, and experimental variances are introduced that can obscure functional distinctions in closely related systems. Here, to address these limitations, we introduce parallel rapid exchange single-molecule fluorescence and single-molecule Frster resonance energy transfer to enable simultaneous steady-state and pre-steady-state interrogations of diverse systems. Using this approach, we elucidate the timing of distinct conformational events underpinning β-arrestin1 activation, unmask antibiotic-induced impacts on messenger RNA decoding fidelity and demonstrate that endogenously encoded ribosomal RNA sequence variation modulates antibiotic sensitivity. This generalizable and scalable method promises to broaden the scope and reproducibility of quantitative single-molecule interrogations of biomolecular function.

DOI: 10.1038/s41592-025-02944-4

Source: https://www.nature.com/articles/s41592-025-02944-4