Many membrane-bound proteins, including cytokines, receptors, and growth factors, are proteolytically cleaved to release a soluble form of their extracellular domain. The tumor necrosis factor (TNF)-α converting enzyme (TACE/ADAM-17) is a transmembrane metalloproteinase responsible for the proteolytic release or 'shedding' of several cell-surface proteins, including TNF and p75 TNFR. We established a TACE-reconstitution system using TACE- deficient cells co-transfected with TACE and substrate cDNAs to study TACE function and regulation. Using the TACE-reconstitution system, we identified two additional substrates of TACE, interleukin (IL)-1R-II and p55 TNFR. Using truncations and chimeric constructs of TACE and another ADAM family member, ADAM-10, we studied the function of the different domains of TACE in three shedding activities. We found that TACE must be expressed with its membrane- anchoring domain for phorbol ester-stimulated shedding of TNF, p75 TNFR, and IL-1R-II, but that the cytoplasmic domain is not required for the shedding of these substrates. The catalytic domain of ADAM-10 could not be functionally substituted for that of TACE. IL-1R-II shedding required the cysteine-rich domain of TACE as well as the catalytic domain, whereas TNF and p75 TNFR shedding required only the tethered TACE catalytic domain.
CITATION STYLE
Reddy, P., Slack, J. L., Davis, R., Cerretti, D. P., Kozlosky, C. J., Blanton, R. A., … Black, R. A. (2000). Functional analysis of the domain structure of tumor necrosis factor-α converting enzyme. Journal of Biological Chemistry, 275(19), 14608–14614. https://doi.org/10.1074/jbc.275.19.14608
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