A cell-based assay for rapid assessment of ACE2 catalytic function.

Autor: Meyers WM; Djavad Mowafaghian Centre for Brain Health, University of British Columbia, Vancouver, BC, Canada.; Department of Cellular & Physiological Sciences, University of British Columbia, Vancouver, BC, Canada., Hong RJ; Djavad Mowafaghian Centre for Brain Health, University of British Columbia, Vancouver, BC, Canada.; Department of Cellular & Physiological Sciences, University of British Columbia, Vancouver, BC, Canada., Sin WC; Djavad Mowafaghian Centre for Brain Health, University of British Columbia, Vancouver, BC, Canada.; Department of Cellular & Physiological Sciences, University of British Columbia, Vancouver, BC, Canada., Kim CS; Djavad Mowafaghian Centre for Brain Health, University of British Columbia, Vancouver, BC, Canada.; Department of Cellular & Physiological Sciences, University of British Columbia, Vancouver, BC, Canada., Haas K; Djavad Mowafaghian Centre for Brain Health, University of British Columbia, Vancouver, BC, Canada. kurt.haas@ubc.ca.; Department of Cellular & Physiological Sciences, University of British Columbia, Vancouver, BC, Canada. kurt.haas@ubc.ca.; School of Biomedical Engineering, University of British Columbia, Vancouver, BC, Canada. kurt.haas@ubc.ca.
Jazyk: angličtina
Zdroj: Scientific reports [Sci Rep] 2023 Aug 29; Vol. 13 (1), pp. 14123. Date of Electronic Publication: 2023 Aug 29.
DOI: 10.1038/s41598-023-41389-7
Abstrakt: Angiotensin-converting enzyme II (ACE2) is a monocarboxypeptidase expressed throughout multiple tissues and its catalysis of bioactive peptides regulates the renin-angiotensin system mediating blood pressure homeostasis. ACE2 is implicated in a variety of diseases, including obesity, diabetes, and cardiovascular diseases, and is the obligate entry receptor for SARS-CoV-2 infection. Disease-associated genetic variants of ACE2 are increasingly being identified but are poorly characterized. To aid this problem, we introduce a fluorometric cell-based assay for evaluating surface-expressed ACE2 catalytic activity that preserves the native glycosylation of the host environment and is amenable to high-throughput analysis of ACE2 variants in multi-well plates. We demonstrate sensitivity to detecting catalysis of the key ACE2 substrates, Angiotensin II, Apelin-13, and des-Arg 9 -bradykinin, and impact of a catalytically-deficient ACE2 variant. Normalizing catalytic measures to surface ACE2 expression accounts for variability in ACE2 variant transfection, surface delivery or stability. This assay provides a convenient and powerful approach for investigating the catalytic characteristics of ACE2 variants involved in cardiovascular peptide cascades and homeostasis of multiple organs.
(© 2023. Springer Nature Limited.)
Databáze: MEDLINE
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