Hydrogen peroxide and endothelin-1 are novel activators of betacellulin ectodomain shedding.
Hydrogen peroxide and endothelin-1 are novel activators of betacellulin ectodomain shedding.
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过氧化氢和内皮素-1 是β细胞素胞外域脱落的新型激活剂。
DOI：
10.1002/jcb.20968
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发表时间：
2006
影响因子：
4
通讯作者：
Dunbar,AndrewJ
中科院分区：
文献类型：
--
作者：
Sanderson,MichaelP;Abbott,CatherineA;Tada,Hiroko;Seno,Masaharu;Dempsey,PeterJ;Dunbar,AndrewJ
betacellulin前体（pro - BTC）是ADAM10介导的外畴脱落的一种新型底物。在本报告中，我们研究了新的生理相关刺激的能力，包括G蛋白偶联受体（GPCR）激动剂和活性氧（ROS），以促进前BTC脱落。我们发现，在过表达pro - BTC的乳腺腺癌MCF7细胞中，过氧化氢（H2O2）是一种强大的外畴脱落刺激剂。h2o2对pro - BTC脱落的刺激被广谱金属蛋白酶抑制剂TAPI‐0阻断，但在ADAM17 （TACE）缺乏的胃上皮细胞中仍然起作用，这表明有一种不同的金属蛋白酶参与。在抗氧化剂N -乙酰- L -半胱氨酸中共培养细胞可以阻断H2O2诱导的前BTC脱落，但在缺钙培养基中培养则不受影响。相比之下，钙离子载体是一种前BTC脱落的激活剂，它对钙的消耗很敏感，但不受与抗氧化剂共培养的影响，这表明了这些刺激之间的明显区别。我们发现，在过表达pro - BTC的血管平滑肌细胞中，GPCR激动剂内皮素- 1 （ET - 1）是外畴脱落的强诱导剂。这被金属蛋白酶抑制剂和催化活性不强的E385A ADAM10过表达阻断。然而，野生型ADAM10或ADAM17的过表达导致ET - 1诱导的前BTC脱落增加，这为这两种酶参与这一过程提供了证据。本研究发现ROS和ET - 1是亲BTC脱落的两种新型诱导剂，并支持在生理相关刺激控制下发生激活脱落的概念。j .细胞。生物化学学报，26(3):693 - 693,2006。©2006 Wiley‐Liss, Inc。
The betacellulin precursor (pro‐BTC) is a novel substrate for ADAM10‐mediated ectodomain shedding. In this report, we investigated the ability of novel physiologically relevant stimuli, including G‐protein coupled receptor (GPCR) agonists and reactive oxygen species (ROS), to stimulate pro‐BTC shedding. We found that in breast adenocarcinoma MCF7 cells overexpressing pro‐BTC, hydrogen peroxide (H2O2) was a powerful stimulator of ectodomain shedding. The stimulation of pro‐BTC shedding by H2O2was blocked by the broad‐spectrum metalloprotease inhibitor TAPI‐0 but was still functional in ADAM17 (TACE)‐deficient stomach epithelial cells indicating the involvement of a distinct metalloprotease. H2O2‐induced pro‐BTC shedding was blocked by co‐culturing cells in the anti‐oxidant N‐acetyl‐L‐cysteine but was unaffected by culture in calcium‐deficient media. By contrast, calcium ionophore, which is a previously characterized activator of pro‐BTC shedding, was sensitive to calcium depletion but was unaffected by co‐culture with the anti‐oxidant, identifying a clear distinction between these stimuli. We found that in vascular smooth muscle cells overexpressing pro‐BTC, the GPCR agonist endothelin‐1 (ET‐1) was a strong inducer of ectodomain shedding. This was blocked by a metalloprotease inhibitor and by overexpression of catalytically inactive E385A ADAM10. However, overexpression of wild‐type ADAM10 or ADAM17 led to an increase in ET‐1‐induced pro‐BTC shedding providing evidence for an involvement of both enzymes in this process. This study identifies ROS and ET‐1 as two novel inducers of pro‐BTC shedding and lends support to the notion of activated shedding occurring under the control of physiologically relevant stimuli. J. Cell. Biochem. 99: 609–623, 2006. © 2006 Wiley‐Liss, Inc.