Exploring extracellular vesicles biogenesis in hypothalamic cells through a heavy isotope pulse/trace proteomic approach
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 de...
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sg-ntu-dr.10356-1447462023-02-28T16:58:09Z Exploring extracellular vesicles biogenesis in hypothalamic cells through a heavy isotope pulse/trace proteomic approach Tan, Chee Fan Teo, Hui San Park, Jung Eun Dutta, Bamaprasad Tse, Shun Wilford Leow, Melvin Khee-Shing Wahli, Walter Sze, Siu Kwan School of Biological Sciences Lee Kong Chian School of Medicine (LKCMedicine) Interdisciplinary Graduate School (IGS) NTU Institute for Health Technologies Science::Medicine Extracellular Vesicles Extracellular Vesicles Biogenesis 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. Ministry of Education (MOE) National Medical Research Council (NMRC) Accepted version This work is in part supported by grants from the Singapore Ministry of Education (MOE2018-T1-001-078and MOE2016-T2-2-018) and the National Medical Research Council of Singapore (NMRC-OF-IRG-0003-2016) 2020-11-23T05:49:02Z 2020-11-23T05:49:02Z 2020 Journal Article 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 0092-8674 https://hdl.handle.net/10356/144746 10.3390/cells9051320 32466345 5 9 en Cells © 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 (http://creativecommons.org/licenses/by/4.0/) application/pdf |
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Science::Medicine Extracellular Vesicles Extracellular Vesicles Biogenesis Tan, Chee Fan Teo, Hui San Park, Jung Eun Dutta, Bamaprasad Tse, Shun Wilford Leow, Melvin Khee-Shing Wahli, Walter Sze, Siu Kwan Exploring extracellular vesicles biogenesis in hypothalamic cells through a heavy isotope pulse/trace proteomic approach |
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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. |
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School of Biological Sciences |
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School of Biological Sciences Tan, Chee Fan Teo, Hui San Park, Jung Eun Dutta, Bamaprasad Tse, Shun Wilford Leow, Melvin Khee-Shing Wahli, Walter Sze, Siu Kwan |
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Article |
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Tan, Chee Fan Teo, Hui San Park, Jung Eun Dutta, Bamaprasad Tse, Shun Wilford Leow, Melvin Khee-Shing Wahli, Walter Sze, Siu Kwan |
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Tan, Chee Fan |
title |
Exploring extracellular vesicles biogenesis in hypothalamic cells through a heavy isotope pulse/trace proteomic approach |
title_short |
Exploring extracellular vesicles biogenesis in hypothalamic cells through a heavy isotope pulse/trace proteomic approach |
title_full |
Exploring extracellular vesicles biogenesis in hypothalamic cells through a heavy isotope pulse/trace proteomic approach |
title_fullStr |
Exploring extracellular vesicles biogenesis in hypothalamic cells through a heavy isotope pulse/trace proteomic approach |
title_full_unstemmed |
Exploring extracellular vesicles biogenesis in hypothalamic cells through a heavy isotope pulse/trace proteomic approach |
title_sort |
exploring extracellular vesicles biogenesis in hypothalamic cells through a heavy isotope pulse/trace proteomic approach |
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2020 |
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https://hdl.handle.net/10356/144746 |
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