Investigation into the Mechanism of Homo- and Heterodimerization of Angiotensin-Converting Enzyme

J Albert Abrie, Wessel J A Moolman, Gyles E Cozier, Sylva L U Schwager, K Ravi Acharya, Edward D Sturrock

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Abstract

Angiotensin-converting enzyme (ACE) plays a central role in the renin-angiotensin system (RAS), which is primarily responsible for blood pressure homeostasis. Studies have shown that ACE inhibitors yield cardiovascular benefits that cannot be entirely attributed to the inhibition of ACE catalytic activity. It is possible that these benefits are due to interactions between ACE and RAS receptors that mediate the protective arm of the RAS, such as the angiotensin-II receptor type 2 (AT2R) and the receptor MAS. Therefore, in this study we investigated the molecular interactions of ACE, including ACE homodimerization and heterodimerization with AT2R and MAS respectively. Molecular interactions were assessed by fluorescence resonance energy transfer and bi-molecular fluorescence complementation in human embryonic kidney-293 cells and Chinese hamster ovary-K1 cells transfected with vectors encoding fluorophore-tagged proteins. The specificity of dimerization was verified by competition experiments using untagged proteins. These techniques were used to study several potential requirements for testis ACE (tACE) dimerization as well as the effect of ACE inhibitors on both somatic ACE (sACE) and tACE dimerization. We demonstrated constitutive homodimerization of somatic ACE and of both its domains separately, as well as heterodimerization of both sACE and tACE with AT2R, but not MAS. Additionally, we investigated both soluble sACE and sACE N-domain using size exclusion chromatography small-angle X-ray scattering and we observed dimers in solution for both forms of the enzyme. Our results suggest that ACE homo- and heterodimerization does occur under physiological conditions.

LanguageEnglish
Pages344-354
JournalMolecular Pharmacology
Volume93
Issue number4
Early online date25 Jan 2018
DOIs
StatusPublished - 1 Apr 2018

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Hominidae
Peptidyl-Dipeptidase A
Dimerization
Renin-Angiotensin System
Testis
Angiotensin-Converting Enzyme Inhibitors
Angiotensin Type 2 Receptor
Fluorescence Resonance Energy Transfer
Angiotensin Receptors
Cricetulus
Gel Chromatography
Ovary
Proteins
Homeostasis
Fluorescence
X-Rays
Blood Pressure
Kidney
Enzymes

Keywords

  • Journal Article

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Investigation into the Mechanism of Homo- and Heterodimerization of Angiotensin-Converting Enzyme. / Abrie, J Albert; Moolman, Wessel J A; Cozier, Gyles E; Schwager, Sylva L U; Acharya, K Ravi; Sturrock, Edward D.

In: Molecular Pharmacology, Vol. 93, No. 4, 01.04.2018, p. 344-354.

Research output: Contribution to journalArticle

Abrie, J Albert ; Moolman, Wessel J A ; Cozier, Gyles E ; Schwager, Sylva L U ; Acharya, K Ravi ; Sturrock, Edward D. / Investigation into the Mechanism of Homo- and Heterodimerization of Angiotensin-Converting Enzyme. In: Molecular Pharmacology. 2018 ; Vol. 93, No. 4. pp. 344-354.
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