Immobilization of Recombinant Fluorescent Biosensors Permits Imaging of Extracellular Ion Signals.
FRET
GEPII 1.0
biotin-traptavidin
cell surface immobilization
extracellular ion measurements
pH-Lemon
recombinant fluorescent biosensors
Journal
ACS sensors
ISSN: 2379-3694
Titre abrégé: ACS Sens
Pays: United States
ID NLM: 101669031
Informations de publication
Date de publication:
26 11 2021
26 11 2021
Historique:
pubmed:
10
11
2021
medline:
15
12
2021
entrez:
9
11
2021
Statut:
ppublish
Résumé
Given the importance of ion gradients and fluxes in biology, monitoring ions locally at the exterior of the plasma membrane of intact cells in a noninvasive manner is highly desirable but challenging. Classical targeting of genetically encoded biosensors at the exterior of cell surfaces would be a suitable approach; however, it often leads to intracellular accumulation of the tools in vesicular structures and adverse modifications, possibly impairing sensor functionality. To tackle these issues, we generated recombinant fluorescent ion biosensors fused to traptavidin (TAv) specifically coupled to a biotinylated AviTag expressed on the outer cell surface of cells. We show that purified chimeras of TAv and pH-Lemon or GEPII 1.0, Förster resonance energy transfer-based pH and K
Identifiants
pubmed: 34752056
doi: 10.1021/acssensors.1c01369
pmc: PMC8630794
doi:
Substances chimiques
Ions
0
Types de publication
Journal Article
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
3994-4000Subventions
Organisme : Austrian Science Fund FWF
ID : I 3716
Pays : Austria
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