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A cell-based assay for rapid assessment of ACE2 catalytic function.
Warren M Meyers1,2, Ryan J Hong1,2, Wun Chey Sin1,2
1Djavad Mowafaghian Centre for Brain Health, University of British Columbia, Vancouver, BC, Canada.
Scientific Reports
|August 29, 2023
Summary
We developed a new assay to measure the activity of Angiotensin-converting enzyme II (ACE2) variants. This tool helps researchers understand ACE2
Area of Science:
- Biochemistry
- Molecular Biology
- Physiology
Background:
- Angiotensin-converting enzyme II (ACE2) is a key enzyme regulating blood pressure and involved in various diseases.
- ACE2 is also the primary entry receptor for SARS-CoV-2.
- Genetic variants of ACE2 are linked to diseases but remain poorly understood.
Purpose of the Study:
- To develop a high-throughput, cell-based assay for evaluating the catalytic activity of surface-expressed ACE2 variants.
- To characterize the functional impact of disease-associated ACE2 variants.
- To provide a tool for investigating ACE2's role in cardiovascular and organ homeostasis.
Main Methods:
- A fluorometric, cell-based assay was designed to measure ACE2 catalytic activity.
- The assay preserves native glycosylation and is suitable for multi-well plate analysis.
- Catalytic activity was normalized to surface ACE2 expression to account for variability.
Main Results:
- The assay successfully detected the catalysis of key ACE2 substrates: Angiotensin II, Apelin-13, and des-Arg9-bradykinin.
- The assay identified the impact of a catalytically deficient ACE2 variant.
- Normalization to surface expression improved the reliability of variant characterization.
Conclusions:
- The developed assay is a sensitive and robust method for assessing ACE2 variant activity.
- This tool facilitates the investigation of ACE2's role in cardiovascular peptide regulation and organ homeostasis.
- Understanding ACE2 variants is crucial for disease research and therapeutic development.
