Hydrogen peroxide and endothelin-1 are novel activators of betacellulin ectodomain shedding

Michael P. Sanderson, Catherine A. Abbott, Hiroko Tada, Masaharu Seno, Peter J. Dempsey, Andrew J. Dunbar

Research output: Contribution to journalArticle

20 Citations (Scopus)

Abstract

The betacellulin precursor (pro-BTC) is a novel substrate for ADAM10-mediated ectodomain shedding. In this report, we investigated the ability of novel physiologically relevant stimuli, including G-protein coupled receptor (GPCR) agonists and reactive oxygen species (ROS), to stimulate pro-BTC shedding. We found that in breast adenocarcinoma MCF7 cells overexpressing pro-BTC, hydrogen peroxide (H2O2) was a powerful stimulator of ectodomain shedding. The stimulation of pro-BTC shedding by H 2O2 was blocked by the broad-spectrum metalloprotease inhibitor TAPI-0 but was still functional in ADAM17 (TACE)-deficient stomach epithelial cells indicating the involvement of a distinct metalloprotease. H2O2-induced pro-BTC shedding was blocked by co-culturing cells in the anti-oxidant N-acetyl-L-cysteine but was unaffected by culture in calcium-deficient media. By contrast, calcium ionophore, which is a previously characterized activator of pro-BTC shedding, was sensitive to calcium depletion but was unaffected by co-culture with the anti-oxidant, identifying a clear distinction between these stimuli. We found that in vascular smooth muscle cells overexpressing pro-BTC, the GPCR agonist endothelin-1 (ET-1) was a strong inducer of ectodomain shedding. This was blocked by a metalloprotease inhibitor and by overexpression of catalytically inactive E385A ADAM10. However, overexpression of wild-type ADAM10 or ADAM17 led to an increase in ET-1-induced pro-BTC shedding providing evidence for an involvement of both enzymes in this process. This study identifies ROS and ET-1 as two novel inducers of pro-BTC shedding and lends support to the notion of activated shedding occurring under the control of physiologically relevant stimuli.

Original languageEnglish
Pages (from-to)609-623
Number of pages15
JournalJournal of Cellular Biochemistry
Volume99
Issue number2
DOIs
Publication statusPublished - Oct 1 2006

Fingerprint

Metalloproteases
Endothelin-1
Hydrogen Peroxide
G-Protein-Coupled Receptors
Oxidants
Reactive Oxygen Species
Calcium
Calcium Ionophores
MCF-7 Cells
Acetylcysteine
Coculture Techniques
Vascular Smooth Muscle
Smooth Muscle Myocytes
Muscle
Stomach
Adenocarcinoma
Breast
Epithelial Cells
Cells
Substrates

Keywords

  • ADAM
  • Betacellulin
  • Ectodomain shedding
  • Endothelin
  • ErbB
  • Metalloprotease
  • Reactive oxygen species

ASJC Scopus subject areas

  • Biochemistry
  • Cell Biology

Cite this

Hydrogen peroxide and endothelin-1 are novel activators of betacellulin ectodomain shedding. / Sanderson, Michael P.; Abbott, Catherine A.; Tada, Hiroko; Seno, Masaharu; Dempsey, Peter J.; Dunbar, Andrew J.

In: Journal of Cellular Biochemistry, Vol. 99, No. 2, 01.10.2006, p. 609-623.

Research output: Contribution to journalArticle

Sanderson, Michael P. ; Abbott, Catherine A. ; Tada, Hiroko ; Seno, Masaharu ; Dempsey, Peter J. ; Dunbar, Andrew J. / Hydrogen peroxide and endothelin-1 are novel activators of betacellulin ectodomain shedding. In: Journal of Cellular Biochemistry. 2006 ; Vol. 99, No. 2. pp. 609-623.
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