Degradome of soluble ADAM10 and ADAM17 metalloproteases
Degradome of soluble ADAM10 and ADAM17 metalloproteases
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DOI：
10.1007/s00018-019-03184-4
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发表时间：
2020-01-01
影响因子：
8
通讯作者：
Becker-Pauly, Christoph
中科院分区：
文献类型：
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作者：
Scharfenberg, Franka;Helbig, Andreas;Becker-Pauly, Christoph
去整合素和金属蛋白酶(ADAMS)10和17可以通过胞外结构域脱落释放多种膜结合蛋白的胞外部分，对许多生物学功能至关重要。到目前为止，底物鉴定主要集中在膜锚定的ADAM10和ADAM17上。然而，除了已知的ADAM10的脱落外，我们还发现ADAM8是一种能够释放ADAM17胞外结构域的蛋白酶。因此，我们研究了ADAM10/17(sADAM10/17)的可溶性胞外区与其膜结合的对应物相比是否显示出改变的底物光谱。基于质谱学的N-末端组学方法确定了小鼠心肌细胞分泌组中总共134个蛋白质切割事件和45个共同的sADAM10/17底物。对这些裂解位点的分析证实了先前发现的氨基酸偏好。进一步的体外研究证实纤维连接蛋白、胱抑素C、SN-钙粘蛋白、PCPE-1以及Sapp是sADAM10和/或sADAM17的直接底物。总体而言，我们首次对sADAM10/17进行了降解组研究，从而引入了一种新的蛋白水解酶活性模式。
Disintegrin and metalloproteinases (ADAMs) 10 and 17 can release the extracellular part of a variety of membrane-bound proteins via ectodomain shedding important for many biological functions. So far, substrate identification focused exclusively on membrane-anchored ADAM10 and ADAM17. However, besides known shedding of ADAM10, we identified ADAM8 as a protease capable of releasing the ADAM17 ectodomain. Therefore, we investigated whether the soluble ectodomains of ADAM10/17 (sADAM10/17) exhibit an altered substrate spectrum compared to their membrane-bound counterparts. A mass spectrometry-based N-terminomics approach identified 134 protein cleavage events in total and 45 common substrates for sADAM10/17 within the secretome of murine cardiomyocytes. Analysis of these cleavage sites confirmed previously identified amino acid preferences. Further in vitro studies verified fibronectin, cystatin C, sN-cadherin, PCPE-1 as well as sAPP as direct substrates of sADAM10 and/or sADAM17. Overall, we present the first degradome study for sADAM10/17, thereby introducing a new mode of proteolytic activity within the protease web.