1. Identification of ADAM10 as a major TNF sheddase in ADAM17-deficient fibroblasts.
- Author
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Mezyk-Kopeć R, Bzowska M, Stalińska K, Chełmicki T, Podkalicki M, Jucha J, Kowalczyk K, Mak P, and Bereta J
- Subjects
- ADAM Proteins genetics, ADAM10 Protein, ADAM17 Protein, Amyloid Precursor Protein Secretases genetics, Animals, Cells, Cultured, Fibroblasts cytology, Humans, Membrane Proteins genetics, Mice, Mice, Knockout, ADAM Proteins metabolism, Amyloid Precursor Protein Secretases metabolism, Fibroblasts physiology, Membrane Proteins metabolism, Tumor Necrosis Factor-alpha metabolism
- Abstract
ADAM17 (a disintegrin and metalloprotease)-deficient murine fibroblasts stably transfected with proTNF cDNA release significant amounts of biologically active soluble TNF. The enzyme responsible for this activity is a membrane protein that hydrolyzes the peptide bond Ala(76):Val(77) within proTNF. Its activity is inhibited by 1,10-phenantroline and GM6001, insusceptible to TIMP-2 (tissue inhibitor of metalloproteinases-2), and stimulated by ionomycin. These characteristics match ADAM10. The moderate silencing of ADAM10 by shRNA resulted in a significant inhibition of TNF shedding. There was no correlation between the level of ADAM10 expression and the presence of active ADAM17. Our results indicate that ADAM10 may function as the TNF sheddase in cells which lack ADAM17 activity.
- Published
- 2009
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