Evaluation of the contribution of different ADAMs to tumor necrosis factor α (TNFα) shedding and of the function of the TNFα ectodomain in ensuring selective stimulated shedding by the TNFα convertase (TACE/ADAM17) Journal Article
| Authors: | Zheng, Y.; Saftig, P.; Hartmann, D.; Blobel, C. |
| Article Title: | Evaluation of the contribution of different ADAMs to tumor necrosis factor α (TNFα) shedding and of the function of the TNFα ectodomain in ensuring selective stimulated shedding by the TNFα convertase (TACE/ADAM17) |
| Abstract: | Tumor necrosis factor-α (TNFα), a potent pro-inflammatory cytokine, is released from cells by proteolytic cleavage of a membrane-anchored precursor. The TNF-α converting enzyme (TACE; a disintegrin and metalloprotease17; ADAM17) is known to have a key role in the ectodomain shedding of TNFα in several cell types. However, because purified ADAMs 9, 10, and 19 can also cleave a peptide corresponding to the TNFα cleavage site in vitro, these enzymes are considered to be candidate TNFα sheddases as well. In this study we used cells lacking ADAMs 9, 10, 17 (TACE), or 19 to address the relative contribution of these ADAMs to TNFα shedding in cell-based assays. Our results corroborate that ADAM17, but not ADAM9, -10, or -19, is critical for phorbol ester- and pervanadate-stimulated release of TNFα in mouse embryonic fibroblasts. However, overexpression of ADAM19 increased the constitutive release of TNFα, whereas overexpression of ADAM9 or ADAM10 did not. This suggests that ADAM19 may contribute to TNFα shedding, especially in cells or tissues where it is highly expressed. Furthermore, we used mutagenesis of TNFα to explore which domains are important for its stimulated processing by ADARI17. We found that the cleavage site of TNFα is necessary and sufficient for cleavage by ADAM17. In addition, the ectodomain of TNFα makes an unexpected contribution to the selective cleavage of TNFα by ADAM17: it prevents one or more other enzymes from cleaving TNFα following PMA stimulation. Thus, selective stimulated processing of TNFα by ADAMI7 in cells depends on the presence of an appropriate cleavage site as well as the inhibitory role of the TNF ectodomain toward other enzymes that can process this site. |
| Keywords: | controlled study; unclassified drug; mutation; nonhuman; protein domain; animal cell; mouse; animals; mice; cells, cultured; gene overexpression; embryo; protein degradation; protein protein interaction; protein binding; membrane proteins; peptide; transfection; cell type; animalia; genetic vectors; blotting, western; cytokine; tumor necrosis factor alpha; tumor necrosis factor-alpha; evaluation; tumors; peptides; immunoprecipitation; fibroblasts; protein structure, tertiary; binding sites; cytokine release; biochemistry; mutagenesis; enzymes; adam proteins; esters; phorbol ester; enzyme; phorbol 13 acetate 12 myristate; cho cells; cricetinae; cells; protein adam 10; tumor necrosis factor alpha converting enzyme; living systems studies; protein precursor; alcohols; disintegrins; metalloendopeptidases; priority journal; article; converting enzymes; proteolytic cleavage; tumor necrosis factor (tnf); pervanadate; protein adam 19; protein adam 9 |
| Journal Title: | Journal of Biological Chemistry |
| Volume: | 279 |
| Issue: | 41 |
| ISSN: | 0021-9258 |
| Publisher: | American Society for Biochemistry and Molecular Biology |
| Date Published: | 2004-10-08 |
| Start Page: | 42898 |
| End Page: | 42906 |
| Language: | English |
| DOI: | 10.1074/jbc.M403193200 |
| PROVIDER: | scopus |
| PUBMED: | 15292243 |
| DOI/URL: | |
| Notes: | J. Biol. Chem. -- Cited By (since 1996):107 -- Export Date: 16 June 2014 -- CODEN: JBCHA -- Source: Scopus |
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