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

Chensu Wang, Yiguang Wang, Yang Li, Brian Bodemann, Tian Zhao, Xinpeng Ma, Gang Huang, Zeping Hu, Ralph J. DeBerardinis, Michael A. White () and Jinming Gao ()
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Chensu Wang: Simmons Comprehensive Cancer Center, University of Texas Southwestern Medical Center
Yiguang Wang: Simmons Comprehensive Cancer Center, University of Texas Southwestern Medical Center
Yang Li: Simmons Comprehensive Cancer Center, University of Texas Southwestern Medical Center
Brian Bodemann: University of Texas Southwestern Medical Center
Tian Zhao: Simmons Comprehensive Cancer Center, University of Texas Southwestern Medical Center
Xinpeng Ma: Simmons Comprehensive Cancer Center, University of Texas Southwestern Medical Center
Gang Huang: Simmons Comprehensive Cancer Center, University of Texas Southwestern Medical Center
Zeping Hu: Children’s Medical Center Research Institute, University of Texas Southwestern Medical Center
Ralph J. DeBerardinis: Children’s Medical Center Research Institute, University of Texas Southwestern Medical Center
Michael A. White: University of Texas Southwestern Medical Center
Jinming Gao: Simmons Comprehensive Cancer Center, University of Texas Southwestern Medical Center

Nature Communications, 2015, vol. 6, issue 1, 1-11

Abstract: Abstract 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.

Date: 2015
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DOI: 10.1038/ncomms9524

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