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

Details

Serval ID
serval:BIB_38F5E271C09D
Type
Article: article from journal or magazin.
Collection
Publications
Institution
Title
Exploring Extracellular Vesicles Biogenesis in Hypothalamic Cells through a Heavy Isotope Pulse/Trace Proteomic Approach.
Journal
Cells
Author(s)
Tan C.F., Teo H.S., Park J.E., Dutta B., Tse S.W., Leow M.K., Wahli W., Sze S.K.
ISSN
2073-4409 (Electronic)
ISSN-L
2073-4409
Publication state
Published
Issued date
25/05/2020
Peer-reviewed
Oui
Volume
9
Number
5
Pages
1320.
Language
english
Notes
Publication types: Journal Article ; Research Support, Non-U.S. Gov't
Publication Status: epublish
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.
Keywords
Animals, Cathepsins/metabolism, Cell Line, Cluster Analysis, Endosomal Sorting Complexes Required for Transport/metabolism, Extracellular Vesicles/metabolism, Extracellular Vesicles/ultrastructure, Gene Ontology, Hypothalamus/cytology, Isotopes/metabolism, Lysosomes/metabolism, Mice, Protein Biosynthesis, Proteome/metabolism, Proteomics/methods, cathepsin, energy homeostasis, extracellular vesicles, extracellular vesicles biogenesis, hypothalamus, pulsed-SILAC
Pubmed
Web of science
Open Access
Yes
Create date
10/06/2020 22:28
Last modification date
28/05/2021 6:36
Usage data