Ectodomain shedding of TNF-α is enhanced by nardilysin via activation of ADAM proteases
- 作者：Yoshinori Hiraoka ; Kazuhiro Yoshida ; Mikiko Ohno ; Tatsuhiko Matsuoka ; Toru Kita ; Eiichiro Nishi
- 关键词：Nardilysin ; Ectodomain shedding ; Tumor necrosis factor-
//www.sciencedirect.com/scidirimg/entities/204e.gif" alt="greek small letter alpha" title="greek small letter alpha" border="0"> (TNF-
- 刊名：Biochemical and Biophysical Research Communications
- 出版年：2008
- 期刊代码：159_0006291x
- 类别：bio
- 出版时间：23 May 2008
- 卷：370
- 期：1
- 页码：154-158
- 文件大小：469 K
摘要
Tumor necrosis factor-
(TNF-) is released from cells by proteolytic cleavage of a membrane-anchored precursor. The TNF--converting enzyme (TACE/ADAM17) is the major sheddase for ectodomain shedding of TNF-. At present, however, it is poorly understood how its catalytic activity is regulated. Here, we show that nardilysin (N-arginine dibasic convertase; NRDc) enhanced TNF- shedding. In a cell-based shedding assay, expression of NRDc synergistically enhanced TACE-induced TNF- shedding. A peptide cleavage assay in vitro showed that recombinant NRDc enhances the cleavage of TNF- induced by TACE. Notably, co-incubation of NRDc completely reversed the inhibitory effect of a physiological concentration of salt on TACE’s activity in vitro. Overexpression of NRDc in TACE-deficient fibroblasts resulted in an increase in the amount of TNF- released. Co-expression of NRDc with ADAM10 promoted the release compared with the sole expression of ADAM10. These results suggested that NRDc enhances TNF- shedding through activation of not only TACE but ADAM10. Our results indicate the involvement of NRDc in ectodomain shedding of TNF-, which may be a novel target for anti-inflammatory therapies.
(TNF-) is released from cells by proteolytic cleavage of a membrane-anchored precursor. The TNF--converting enzyme (TACE/ADAM17) is the major sheddase for ectodomain shedding of TNF-. At present, however, it is poorly understood how its catalytic activity is regulated. Here, we show that nardilysin (N-arginine dibasic convertase; NRDc) enhanced TNF- shedding. In a cell-based shedding assay, expression of NRDc synergistically enhanced TACE-induced TNF- shedding. A peptide cleavage assay in vitro showed that recombinant NRDc enhances the cleavage of TNF- induced by TACE. Notably, co-incubation of NRDc completely reversed the inhibitory effect of a physiological concentration of salt on TACE’s activity in vitro. Overexpression of NRDc in TACE-deficient fibroblasts resulted in an increase in the amount of TNF- released. Co-expression of NRDc with ADAM10 promoted the release compared with the sole expression of ADAM10. These results suggested that NRDc enhances TNF- shedding through activation of not only TACE but ADAM10. Our results indicate the involvement of NRDc in ectodomain shedding of TNF-, which may be a novel target for anti-inflammatory therapies.