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J Biol Chem, Vol. 275, Issue 19, 14598-14607, May 12, 2000
From the Department of Research Administration, Immunex
Corporation, Seattle, Washington 98101
The extracellular domains of many proteins,
including growth factors, cytokines, receptors, and adhesion molecules,
are proteolytically released from cells, a process termed
"shedding." Tumor necrosis factor-
Stimulation-induced Down-regulation of Tumor Necrosis Factor-
Converting Enzyme*
and
converting enzyme
(TACE/ADAM-17) is a metalloprotease-disintegrin that sheds tumor
necrosis factor-
and other proteins. To study the regulation of
TACE-mediated shedding, we examined the effects of stimulation of cells
on TACE localization and expression. Immunofluorescence microscopy
revealed a punctate distribution of TACE on the surface of untreated
cells, and stimulation of monocytic cells with lipopolysaccharide did
not affect TACE staining. Phorbol 12-myristate 13-acetate (PMA), a
potent inducer of shedding, decreased cell-surface staining for TACE.
Surface biotinylation experiments confirmed and extended this
observation; PMA decreased the half-life of surface-biotinylated TACE
without increasing the turnover of total cell-surface proteins. Soluble
fragments of TACE were not detected in the medium of cells that had
down-regulated TACE, and TACE was not down-regulated when endocytosis
was inhibited. Antibody uptake experiments suggested that cell-surface
TACE was internalized in response to PMA. Surprisingly, a
metalloprotease inhibitor prevented the PMA-induced turnover of TACE.
Thus, PMA activates shedding and causes the down-regulation of a major
"sheddase," suggesting that induced shedding may be regulated by a
mechanism that decreases the amount of active TACE on the cell surface.
*
The costs of publication of this
article were defrayed in part by the
payment of page charges. The article
must therefore be hereby marked
"advertisement" in
accordance with 18 U.S.C. Section
1734 solely to indicate this fact.
To whom correspondence should be addressed: Immunex Corp., 51 University St., Seattle, WA 98101. Tel.: 206-587-0430; Fax: 206-621-7977; E-mail: doedensj@immunex.com.
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