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A biosensor for the activity of the “sheddase” TACE (ADAM17) reveals novel and cell type–specific mechanisms of TACE activation

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Sci. Signal.  24 Feb 2015:
Vol. 8, Issue 365, pp. rs1
DOI: 10.1126/scisignal.2005680

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Detecting TACE activity

The ability to visually monitor enzymatic activity in live cells and in real time provides insight on a protein’s spatiotemporal function. Through cleavage, the protease TACE (tumor necrosis factor–α–converting enzyme) releases growth factors, cytokines, and other proteins from the cell surface during processes such as inflammation. Chapnick et al. developed a fluorescence-based sensor called “TSen” that contained the TACE cleavage site, and reported where and when TACE was active. They used TSen to explore the regulation of TACE in several epithelial cell lines. In addition to identifying known activators of TACE, TSen revealed that disruption of the cell’s cytoskeleton also activated TACE. TSen is a valuable tool for investigating the spatial and temporal dynamics of TACE-mediated protein shedding in cells and tissues.


Diverse environmental conditions stimulate protein “shedding” from the cell surface through proteolytic cleavage. The protease TACE [tumor necrosis factor–α (TNFα)–converting enzyme, encoded by ADAM17] mediates protein shedding, thereby regulating the maturation and release of various extracellular substrates, such as growth factors and cytokines, that induce diverse cellular responses. We developed a FRET (fluorescence resonance energy transfer)–based biosensor called TSen that quantitatively reports the kinetics of TACE activity in live cells. In combination with chemical biology approaches, we used TSen to probe the dependence of TACE activation on the induction of the kinases p38 and ERK (extracellular signal–regulated kinase) in various epithelial cell lines. Using TSen, we found that disruption of the actin cytoskeleton in keratinocytes induced rapid and robust TSen cleavage and the accumulation of TACE at the plasma membrane. Cytoskeletal disruption also increased the cleavage of endogenous TACE substrates, including transforming growth factor–α. Thus, TSen is a useful tool for unraveling the mechanisms underlying the spatiotemporal activation of TACE in live cells.

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