Exploring Extracellular Vesicles Biogenesis in Hypothalamic Cells through a Heavy Isotope Pulse/Trace Proteomic Approach.

Autor:	Tan CF; NTU Institute for Health Technologies, Interdisciplinary Graduate School, Nanyang Technological University, Singapore 637335 Singapore.; School of Biological Sciences, Nanyang Technological University, Singapore 637551, Singapore., Teo HS; School of Biological Sciences, Nanyang Technological University, Singapore 637551, Singapore., Park JE; School of Biological Sciences, Nanyang Technological University, Singapore 637551, Singapore., Dutta B; School of Biological Sciences, Nanyang Technological University, Singapore 637551, Singapore., Tse SW; School of Biological Sciences, Nanyang Technological University, Singapore 637551, Singapore., Leow MK; Lee Kong Chian School of Medicine, Nanyang Technological University, Singapore 636921, Singapore.; Department of Endocrinology, Tan Tock Seng Hospital, Singapore 308433, Singapore.; Cardiovascular and Metabolic Disorder Program, Duke-NUS Medical School, Singapore 169857, Singapore., Wahli W; Lee Kong Chian School of Medicine, Nanyang Technological University, Singapore 636921, Singapore.; Center for Integrative Genomics, University of Lausanne, Le Génopode, CH-1015 Lausanne, Switzerland., Sze SK; School of Biological Sciences, Nanyang Technological University, Singapore 637551, Singapore.
Jazyk:	angličtina
Zdroj:	Cells [Cells] 2020 May 25; Vol. 9 (5). Date of Electronic Publication: 2020 May 25.
DOI:	10.3390/cells9051320
Abstrakt:	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.
Databáze:	MEDLINE
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