Max Planck Institute for Polymer Research, 55128 Mainz, Germany.
J Am Chem Soc. 2024 May 1;146(17):11991-11999. doi: 10.1021/jacs.4c01279. Epub 2024 Apr 19.
The complex dynamics and transience of assembly pathways in living systems complicate the understanding of these molecular to nanoscale processes. Current technologies are unable to track the molecular events leading to the onset of assembly, where real-time information is imperative to correlate their rich biology. Using a chemically designed pro-assembling molecule, we map its transformation into nanofibers and their fusion with endosomes to form hollow fiber clusters. Tracked by phasor-fluorescence lifetime imaging (phasor-FLIM) in epithelial cells (L929, A549, MDA-MB 231) and correlative light-electron microscopy and tomography (CLEM), spatiotemporal splicing of the assembly events shows time-correlated metabolic dysfunction. The biological impact begins with assembly-induced endosomal disruption that reduces glucose transport into the cells, which, in turn, stymies mitochondrial respiration.
在生命系统中，组装途径的复杂动态和短暂性使得理解这些从分子到纳米尺度的过程变得复杂。目前的技术无法跟踪导致组装开始的分子事件，而实时信息对于将它们丰富的生物学联系起来至关重要。我们使用一种化学设计的预组装分子，将其转化为纳米纤维，并将其与内体融合形成空心纤维簇。通过在上皮细胞（L929、A549、MDA-MB 231）中进行相位荧光寿命成像（phasor-FLIM）和相关的光电子显微镜和断层扫描（CLEM）进行跟踪，组装事件的时空拼接显示出与时间相关的代谢功能障碍。生物影响始于组装诱导的内体破坏，从而减少葡萄糖向细胞内的转运，进而抑制线粒体呼吸。
