Interleukin-1 stimulates ADAM17 through a mechanism independent of its cytoplasmic domain or phosphorylation at threonine 735.
Interleukin-1 stimulates ADAM17 through a mechanism independent of its cytoplasmic domain or phosphorylation at threonine 735.
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DOI：
10.1371/journal.pone.0031600
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发表时间：
2012
期刊：
影响因子：
3.7
通讯作者：
Blobel CP
中科院分区：
文献类型：
--
作者：
Hall KC;Blobel CP
ADAM17（一种崩解素和金属蛋白酶）是一种膜锚定的金属蛋白酶，调节egfr配体、TNFα和其他膜蛋白从细胞中释放。ADAM17可以通过多种信号通路快速激活，但对其潜在机制知之甚少。一些研究表明，ADAM17的胞质结构域并不需要被各种刺激快速激活，包括佛波酯、酪氨酸激酶和一些g蛋白偶联受体。然而，细胞质残基T735的磷酸化最近被报道为IL-1β和p38 MAP-kinase途径激活ADAM17的关键步骤。调和这些结果的一种可能机制是T735具有抑制作用，并且必须将其磷酸化作为ADAM17激活的先决条件，然后通过独立于其细胞质结构域的机制进行。为了验证这一假设，我们对Adam17野生型和突变型的Adam17 - / -细胞进行了修复实验。然而，这些实验表明，细胞质残基T735的失活突变（T735A）或激活突变（T735D）或ADAM17细胞质结构域的去除对IL-1β或用异霉素激活map -激酶对ADAM17的刺激没有显著影响。此外，我们发现map -激酶抑制剂SB203580阻断了IL-1β或大霉素对细胞质尾缺陷ADAM17和T735A突变体的激活，进一步支持了ADAM17的激活机制不依赖于其细胞质结构域或T735磷酸化的模型。
ADAM17 (a disintegrin and metalloproteinase) is a membrane-anchored metalloproteinase that regulates the release of EGFR-ligands, TNFα and other membrane proteins from cells. ADAM17 can be rapidly activated by a variety of signaling pathways, yet little is known about the underlying mechanism. Several studies have demonstrated that the cytoplasmic domain of ADAM17 is not required for its rapid activation by a variety of stimuli, including phorbol esters, tyrosine kinases and some G-protein coupled receptors. However, phosphorylation of cytoplasmic residue T735 was recently reported as a crucial step for activation of ADAM17 by IL-1β and by the p38 MAP-kinase pathway. One possible mechanism to reconcile these results would be that T735 has an inhibitory role and that it must be phosphorylated as a pre-requisite for the activation of ADAM17, which would then proceed via a mechanism that is independent of its cytoplasmic domain. To test this hypothesis, we performed rescue experiments of Adam17−/− cells with wild type and mutant forms of ADAM17. However, these experiments showed that an inactivating mutation (T735A) or an activating mutation (T735D) of cytoplasmic residue T735 or the removal of the cytoplasmic domain of ADAM17 did not significantly affect the stimulation of ADAM17 by IL-1β or by activation of MAP-kinase with anisomycin. Moreover, we found that the MAP-kinase inhibitor SB203580 blocked activation of cytoplasmic tail-deficient ADAM17 and of the T735A mutant by IL-1β or by anisomycin, providing further support for a model in which the activation mechanism of ADAM17 does not rely on its cytoplasmic domain or phosphorylation of T735.
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