Exploring Extracellular Vesicles Biogenesis in Hypothalamic Cells through a Heavy Isotope Pulse/Trace Proteomic Approach.
Animals
Cathepsins
/ metabolism
Cell Line
Cluster Analysis
Endosomal Sorting Complexes Required for Transport
/ metabolism
Extracellular Vesicles
/ metabolism
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
Journal
Cells
ISSN: 2073-4409
Titre abrégé: Cells
Pays: Switzerland
ID NLM: 101600052
Informations de publication
Date de publication:
25 05 2020
25 05 2020
Historique:
received:
24
03
2020
revised:
13
05
2020
accepted:
21
05
2020
entrez:
30
5
2020
pubmed:
30
5
2020
medline:
25
2
2021
Statut:
epublish
Résumé
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.
Identifiants
pubmed: 32466345
pii: cells9051320
doi: 10.3390/cells9051320
pmc: PMC7291124
pii:
doi:
Substances chimiques
Endosomal Sorting Complexes Required for Transport
0
Isotopes
0
Proteome
0
Cathepsins
EC 3.4.-
Types de publication
Journal Article
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
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