Functional and molecular evidence for expression of the renin angiotensin system and ADAM17-mediated ACE2 shedding in COS7 cells

American Journal of Physiology. Cell Physiology
Nadja GrobeKhalid M Elased

Abstract

The renin angiotensin system (RAS) plays a vital role in the regulation of the cardiovascular and renal functions. COS7 is a robust and easily transfectable cell line derived from the kidney of the African green monkey, Cercopithecus aethiops. The aims of this study were to 1) demonstrate the presence of an endogenous and functional RAS in COS7, and 2) investigate the role of a disintegrin and metalloproteinase-17 (ADAM17) in the ectodomain shedding of angiotensin converting enzyme-2 (ACE2). Reverse transcription coupled to gene-specific polymerase chain reaction demonstrated expression of ACE, ACE2, angiotensin II type 1 receptor (AT1R), and renin at the transcript levels in total RNA cell extracts. Western blot and immunohistochemistry identified ACE (60 kDa), ACE2 (75 kDa), AT1R (43 kDa), renin (41 kDa), and ADAM17 (130 kDa) in COS7. At the functional level, a sensitive and selective mass spectrometric approach detected endogenous renin, ACE, and ACE2 activities. ANG-(1-7) formation (m/z 899) from the natural substrate ANG II (m/z 1,046) was detected in lysates and media. COS7 cells stably expressing shRNA constructs directed against endogenous ADAM17 showed reduced ACE2 shedding into the media. This is the first study demon...Continue Reading

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Citations

Mar 22, 2018·American Journal of Physiology. Renal Physiology·Sridevi GuttaKhalid M Elased
May 30, 2019·American Journal of Physiology. Renal Physiology·Vanesa PalauMarta Riera
Sep 25, 2015·World Journal of Gastroenterology : WJG·Hanne SalmenkariRiitta Korpela
Jan 20, 2017·Frontiers in Cell and Developmental Biology·Richa SinghMauricio Di Fulvio
Nov 17, 2020·Frontiers in Molecular Biosciences·Federica SaponaroRiccardo Zucchi

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