Click here to close
Hello! We notice that you are using Internet Explorer, which is not supported by Xenbase and may cause the site to display incorrectly.
We suggest using a current version of Chrome,
FireFox, or Safari.
Biosensors (Basel)
2022 Aug 04;128:. doi: 10.3390/bios12080601.
Show Gene links
Show Anatomy links
Fluorescence-Based Measurements of Membrane-Bound Angiotensin Converting Enzyme 2 Activity Using Xenopus Laevis Oocytes.
Fast L
,
Ågren R
,
Zeberg H
.
???displayArticle.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.
2021-03050 Swedish Research Council, FO2020-0289 and FO2021-0400 Stiftelsen Lars Hiertas Minne, 2020-00625 and 2021-00683 Fredrik och Ingrid Thurings Stiftelse, 2020-04055 and 2021-04489 Magnus Bergvalls stiftelse, 213066 Åhlén-stiftelsen
Boland,
Extractive purification of enzymes from animal tissue using aqueous two phase systems: pilot scale studies.
1991, Pubmed
Boland,
Extractive purification of enzymes from animal tissue using aqueous two phase systems: pilot scale studies.
1991,
Pubmed
Bröer,
Xenopus laevis Oocytes.
2010,
Pubmed
,
Xenbase
Dascal,
The use of Xenopus oocytes for the study of ion channels.
1987,
Pubmed
,
Xenbase
Graf,
Fully automated microinjection system for Xenopus laevis oocytes with integrated sorting and collection.
2011,
Pubmed
,
Xenbase
Jespersen,
Dual-function vector for protein expression in both mammalian cells and Xenopus laevis oocytes.
2002,
Pubmed
,
Xenbase
Kim,
Mammalian cell transfection: the present and the future.
2010,
Pubmed
Lee,
Methodological improvements for fluorescence recordings in Xenopus laevis oocytes.
2019,
Pubmed
,
Xenbase
McDonald,
ExplorEnz: the primary source of the IUBMB enzyme list.
2009,
Pubmed
Ooi,
A Guide to Transient Expression of Membrane Proteins in HEK-293 Cells for Functional Characterization.
2016,
Pubmed
Ou,
Methods of Measuring Enzyme Activity Ex Vivo and In Vivo.
2018,
Pubmed
Paucar,
V374A KCND3 Pathogenic Variant Associated With Paroxysmal Ataxia Exacerbations.
2021,
Pubmed
,
Xenbase
Rice,
Evaluation of angiotensin-converting enzyme (ACE), its homologue ACE2 and neprilysin in angiotensin peptide metabolism.
2004,
Pubmed
Sahlholm,
Agonist-specific voltage sensitivity at the dopamine D2S receptor--molecular determinants and relevance to therapeutic ligands.
2011,
Pubmed
,
Xenbase
Touret,
In vitro screening of a FDA approved chemical library reveals potential inhibitors of SARS-CoV-2 replication.
2020,
Pubmed
Vickers,
Hydrolysis of biological peptides by human angiotensin-converting enzyme-related carboxypeptidase.
2002,
Pubmed
Yan,
Structural basis for the recognition of SARS-CoV-2 by full-length human ACE2.
2020,
Pubmed
Ågren,
The Beta-Arrestin-Biased Dopamine D2 Receptor Ligand, UNC9994, Is a Partial Agonist at G-Protein-Mediated Potassium Channel Activation.
2018,
Pubmed
,
Xenbase