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Title: Exploring extracellular vesicles biogenesis in hypothalamic cells through a heavy isotope pulse/trace proteomic approach
Authors: Tan, Chee Fan
Teo, Hui San
Park, Jung Eun
Dutta, Bamaprasad
Tse, Shun Wilford
Leow, Melvin Khee-Shing
Wahli, Walter
Sze, Siu Kwan
Keywords: Science::Medicine
Issue Date: 2020
Source: Tan, C. F., Teo, H. S., Park, J. E., Dutta, B., Tse, S. W., Leow, M. K.-S., . . . Sze, S. K. (2020). Exploring Extracellular Vesicles Biogenesis in Hypothalamic Cells through a Heavy Isotope Pulse/Trace Proteomic Approach. Cells, 9(5), 1320-. doi:10.3390/cells9051320
Journal: Cells
Abstract: Studies have shown that the process of extracellular vesicles (EVs) secretion and lysosome status are linked. When the lysosome is under stress, the cells would secrete more EVs to maintain cellular homeostasis. However, the process that governs lysosomal activity and EVs secretion remains poorly defined and we postulated that certain proteins essential for EVs biogenesis are constantly synthesized and preferentially sorted to the EVs rather than the lysosome. A pulsed stable isotope labelling of amino acids in cell culture (pSILAC) based quantitative proteomics methodology was employed to study the preferential localization of the newly synthesized proteins into the EVs over lysosome in mHypoA 2/28 hypothalamic cell line. Through proteomic analysis, we found numerous newly synthesized lysosomal enzymes-such as the cathepsin proteins-that preferentially localize into the EVs over the lysosome. Chemical inhibition against cathepsin D promoted EVs secretion and a change in the EVs protein composition and therefore indicates its involvement in EVs biogenesis. In conclusion, we applied a heavy isotope pulse/trace proteomic approach to study EVs biogenesis in hypothalamic cells. The results demonstrated the regulation of EVs secretion by the cathepsin proteins that may serve as a potential therapeutic target for a range of neurological disorder associated with energy homeostasis.
ISSN: 0092-8674
DOI: 10.3390/cells9051320
Rights: © 2020 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open accessarticle distributed under the terms and conditions of the Creative Commons Attribution(CC BY) license (
Fulltext Permission: open
Fulltext Availability: With Fulltext
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