Fluorescence-Based Measurements of Membrane-Bound Angiotensin Converting Enzyme 2 Activity Using Xenopus Laevis Oocytes
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...
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| Format: | Article |
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MDPI AG
2022-08-01
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| Series: | Biosensors |
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| Online Access: | https://www.mdpi.com/2079-6374/12/8/601 |
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| author | Luise Fast Richard Ågren Hugo Zeberg |
| author_facet | Luise Fast Richard Ågren Hugo Zeberg |
| author_sort | Luise Fast |
| collection | DOAJ |
| description | 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 <i>ACE2</i> cRNA. These results suggest that Xenopus oocytes can be used for the functional evaluation of membrane-bound enzymes, decreasing the experimental workload. |
| first_indexed | 2024-03-09T04:38:50Z |
| format | Article |
| id | doaj.art-2d57dafa3d44487c917d2be9ab3b9176 |
| institution | Directory Open Access Journal |
| issn | 2079-6374 |
| language | English |
| last_indexed | 2024-03-09T04:38:50Z |
| publishDate | 2022-08-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Biosensors |
| spelling | doaj.art-2d57dafa3d44487c917d2be9ab3b91762023-12-03T13:23:07ZengMDPI AGBiosensors2079-63742022-08-0112860110.3390/bios12080601Fluorescence-Based Measurements of Membrane-Bound Angiotensin Converting Enzyme 2 Activity Using Xenopus Laevis OocytesLuise Fast0Richard Ågren1Hugo Zeberg2Department of Evolutionary Genetics, Max Planck Institute for Evolutionary Anthropology, 04103 Leipzig, GermanyDepartment of Neuroscience, Karolinska Institutet, 17177 Stockholm, SwedenDepartment of Evolutionary Genetics, Max Planck Institute for Evolutionary Anthropology, 04103 Leipzig, GermanyFunctional 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 <i>ACE2</i> cRNA. These results suggest that Xenopus oocytes can be used for the functional evaluation of membrane-bound enzymes, decreasing the experimental workload.https://www.mdpi.com/2079-6374/12/8/601angiotensin converting enzyme 2microplate readerXenopus laevis oocytesfluorescence intensityenzymatic assay |
| spellingShingle | Luise Fast Richard Ågren Hugo Zeberg Fluorescence-Based Measurements of Membrane-Bound Angiotensin Converting Enzyme 2 Activity Using Xenopus Laevis Oocytes Biosensors angiotensin converting enzyme 2 microplate reader Xenopus laevis oocytes fluorescence intensity enzymatic assay |
| title | Fluorescence-Based Measurements of Membrane-Bound Angiotensin Converting Enzyme 2 Activity Using Xenopus Laevis Oocytes |
| title_full | Fluorescence-Based Measurements of Membrane-Bound Angiotensin Converting Enzyme 2 Activity Using Xenopus Laevis Oocytes |
| title_fullStr | Fluorescence-Based Measurements of Membrane-Bound Angiotensin Converting Enzyme 2 Activity Using Xenopus Laevis Oocytes |
| title_full_unstemmed | Fluorescence-Based Measurements of Membrane-Bound Angiotensin Converting Enzyme 2 Activity Using Xenopus Laevis Oocytes |
| title_short | Fluorescence-Based Measurements of Membrane-Bound Angiotensin Converting Enzyme 2 Activity Using Xenopus Laevis Oocytes |
| title_sort | fluorescence based measurements of membrane bound angiotensin converting enzyme 2 activity using xenopus laevis oocytes |
| topic | angiotensin converting enzyme 2 microplate reader Xenopus laevis oocytes fluorescence intensity enzymatic assay |
| url | https://www.mdpi.com/2079-6374/12/8/601 |
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