Fluorescence-Based Measurements of Membrane-Bound Angiotensin Converting Enzyme 2 Activity Using Xenopus Laevis Oocytes.
Xenopus laevis oocytes
angiotensin converting enzyme 2
enzymatic assay
fluorescence intensity
microplate reader
Journal
Biosensors
ISSN: 2079-6374
Titre abrégé: Biosensors (Basel)
Pays: Switzerland
ID NLM: 101609191
Informations de publication
Date de publication:
04 Aug 2022
04 Aug 2022
Historique:
received:
01
07
2022
revised:
01
08
2022
accepted:
02
08
2022
entrez:
25
8
2022
pubmed:
26
8
2022
medline:
27
8
2022
Statut:
epublish
Résumé
Functional investigations of enzymes involving cellular expression systems are important for pharmacological studies. The precise control of expression is challenging in transiently transfected mammalian cell lines. Here, we explored the ability of Xenopus laevis oocytes to express a membrane-bound enzyme for functional characterization using standard 96-well plates and a fluorescence-based plate reader assay. We microinjected oocytes with cRNA encoding the angiotensin converting enzyme 2 (ACE2) and measured the enzymatic activity in single oocytes using a commercial fluorescence-based assay. The injected oocytes showed up to a 50-fold increase in fluorescence compared to uninjected oocytes. This fluorescence intensity was dose-dependent on the amount of
Identifiants
pubmed: 36004997
pii: bios12080601
doi: 10.3390/bios12080601
pmc: PMC9405939
pii:
doi:
Substances chimiques
RNA, Complementary
0
Angiotensin-Converting Enzyme 2
EC 3.4.17.23
Types de publication
Journal Article
Langues
eng
Sous-ensembles de citation
IM
Subventions
Organisme : Swedish Research Council
ID : 2021-03050
Organisme : Stiftelsen Lars Hiertas Minne
ID : FO2020-0289 and FO2021-0400
Organisme : Fredrik och Ingrid Thurings Stiftelse
ID : 2020-00625 and 2021-00683
Organisme : Magnus Bergvalls stiftelse
ID : 2020-04055 and 2021-04489
Organisme : Åhlén-stiftelsen
ID : 213066
Déclaration de conflit d'intérêts
The authors declare no conflict of interest.
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