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XB-ART-59289
Biosensors (Basel) 2022 Aug 04;128:. doi: 10.3390/bios12080601.
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Fluorescence-Based Measurements of Membrane-Bound Angiotensin Converting Enzyme 2 Activity Using Xenopus Laevis Oocytes.

Fast L , Ågren R , Zeberg H .


Abstract
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 ACE2 cRNA. These results suggest that Xenopus oocytes can be used for the functional evaluation of membrane-bound enzymes, decreasing the experimental workload.

PubMed ID: 36004997
PMC ID: PMC9405939
Article link: Biosensors (Basel)
Grant support: [+]

Species referenced: Xenopus laevis
Genes referenced: ace2

References [+] :
Boland, Extractive purification of enzymes from animal tissue using aqueous two phase systems: pilot scale studies. 1991, Pubmed