Supramolecular Assembly in Live Cells Mapped by Real-Time Phasor-Fluorescence Lifetime Imaging

作者：Yong Ren, Zhixuan Zhou, Konrad Maxeiner, A. Kaltbeitzel, Iain Harley, Jiaqi Xing, Yingke Wu, Manfred Wagner, K. Landfester, Ingo Lieberwirth, Tanja Weil, D. Ng · 发表于：Journal of the American Chemical Society · 年份：2024 · DOI：10.1021/jacs.4c01279 · 被引用次数：34 · 研究领域：Medicine

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.