Membrane-bound Gaussia luciferase as a tool to track shedding of membrane proteins from the surface of extracellular vesicles.
Journal
Scientific reports
ISSN: 2045-2322
Titre abrégé: Sci Rep
Pays: England
ID NLM: 101563288
Informations de publication
Date de publication:
22 11 2019
22 11 2019
Historique:
received:
16
08
2019
accepted:
25
10
2019
entrez:
24
11
2019
pubmed:
24
11
2019
medline:
1
12
2020
Statut:
epublish
Résumé
Extracellular vesicles (EVs) released by cells play a role in intercellular communication. Reporter and targeting proteins can be modified and exposed on the surface of EVs to investigate their half-life and biodistribution. A characterization of membrane-bound Gaussia luciferase (mbGluc) revealed that its signal was detected also in a form smaller than common EVs (<70 nm). We demonstrated that mbGluc initially exposed on the surface of EVs, likely undergoes proteolytic cleavage and processed fragments of the protein are released into the extracellular space in active form. Based on this observation, we developed a new assay to quantitatively track shedding of membrane proteins from the surface of EVs. We used this assay to show that ectodomain shedding in EVs is continuous and is mediated by specific proteases, e.g. metalloproteinases. Here, we present a novel tool to study membrane protein cleavage and release using both in vitro and in vivo models.
Identifiants
pubmed: 31758005
doi: 10.1038/s41598-019-53554-y
pii: 10.1038/s41598-019-53554-y
pmc: PMC6874653
doi:
Substances chimiques
Membrane Proteins
0
Recombinant Proteins
0
Luciferases
EC 1.13.12.-
Types de publication
Journal Article
Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
17387Subventions
Organisme : U.S. Department of Health & Human Services | National Institutes of Health (NIH)
ID : R01CA229777
Pays : International
Organisme : U.S. Department of Health & Human Services | National Institutes of Health (NIH)
ID : U01CA233360
Pays : International
Organisme : NCI NIH HHS
ID : R35 CA232103
Pays : United States
Organisme : U.S. Department of Health & Human Services | National Institutes of Health (NIH)
ID : P01CA069246
Pays : International
Organisme : NCI NIH HHS
ID : R01 CA229777
Pays : United States
Organisme : NHLBI NIH HHS
ID : R01 HL127173
Pays : United States
Organisme : NHLBI NIH HHS
ID : R01 HL128996
Pays : United States
Organisme : NINDS NIH HHS
ID : P30 NS045776
Pays : United States
Organisme : NHLBI NIH HHS
ID : P01 HL116263
Pays : United States
Organisme : NCI NIH HHS
ID : U19 CA179563
Pays : United States
Organisme : NCI NIH HHS
ID : U01 CA233360
Pays : United States
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