A nanobuffer reporter library for fine-scale imaging and perturbation of endocytic organelles

• Published in: Nature communications Vol. 6; no. 1; p. 8524
• Main Authors: Wang, Chensu, Wang, Yiguang, Li, Yang, Bodemann, Brian, Zhao, Tian, Ma, Xinpeng, Huang, Gang, Hu, Zeping, DeBerardinis, Ralph J., White, Michael A., Gao, Jinming
• Format: Journal Article
• Language: English
• Published: London Nature Publishing Group UK 05-10-2015; Nature Publishing Group; Nature Pub. Group
• Subjects: 13/1; 13/2; 13/95; 14/19; 14/34; 631/1647/350/354; 631/80/642/1776; 631/92/320; Amino Acids - metabolism; Apoptosis; Carcinoma, Non-Small-Cell Lung - metabolism; Carcinoma, Non-Small-Cell Lung - ultrastructure; Cell Line; Cell Line, Tumor; Endocytosis; Endosomes - metabolism; Endosomes - ultrastructure; Fluorescent Dyes; HeLa Cells; Humanities and Social Sciences; Humans; Hydrogen-Ion Concentration; Lung Neoplasms - metabolism; Lung Neoplasms - ultrastructure; Lysosomes - metabolism; Lysosomes - ultrastructure; Mechanistic Target of Rapamycin Complex 1; Metabolome; Microscopy, Confocal; Microscopy, Fluorescence; multidisciplinary; Multiprotein Complexes - metabolism; Nanoparticles; Organelles - ultrastructure; Proteins - metabolism; Science; Science (multidisciplinary); TOR Serine-Threonine Kinases - metabolism; Vacuoles - ultrastructure
• ISSN: 2041-1723; 2041-1723
• DOI: 10.1038/ncomms9524

Endosomes, lysosomes and related catabolic organelles are a dynamic continuum of vacuolar structures that impact a number of cell physiological processes such as protein/lipid metabolism, nutrient sensing and cell survival. Here we develop a library of ultra-pH-sensitive fluorescent nanoparticles with chemical properties that allow fine-scale, multiplexed, spatio-temporal perturbation and quantification of catabolic organelle maturation at single organelle resolution to support quantitative investigation of these processes in living cells. Deployment in cells allows quantification of the proton accumulation rate in endosomes; illumination of previously unrecognized regulatory mechanisms coupling pH transitions to endosomal coat protein exchange; discovery of distinct pH thresholds required for mTORC1 activation by free amino acids versus proteins; broad-scale characterization of the consequence of endosomal pH transitions on cellular metabolomic profiles; and functionalization of a context-specific metabolic vulnerability in lung cancer cells. Together, these biological applications indicate the robustness and adaptability of this nanotechnology-enabled ‘detection and perturbation’ strategy.
Endocytic organelles are a dynamic continuum of vacuolar structures that impact various cell physiological processes. Here, the authors report a library of ultra-pH sensitive fluorescent nanoparticles for the controlled perturbation and quantification of catabolic organelle maturation at organelle resolution.