Diverse cell surface protein ectodomains are shed by a system sensitive to metalloprotease inhibitors
Diverse cell surface protein ectodomains are shed by a system sensitive to metalloprotease inhibitors
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DOI：
10.1074/jbc.271.19.11376
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发表时间：
1996-05-10
影响因子：
4.8
通讯作者：
Massague, J
中科院分区：
文献类型：
--
作者：
Arribas, J;Coodly, L;Massague, J
一组不同的膜蛋白的胞外结构域响应于蛋白激酶C激活剂如佛波醇12-肉豆蔻酸酯13-乙酸酯（PMA）而脱落。在切割位点中缺乏序列相似性表明在该过程中涉及许多不同特异性的蛋白酶。然而，最近分离出的一种突变型中国仓鼠卵巢细胞系在PMA激活的膜锚定生长因子转化生长因子α前体（proTGF-α）脱落中存在缺陷，同时在许多其他不相关的膜蛋白脱落中也存在缺陷。在这里，我们表明，独立的诱变和选择实验产生脱落突变体具有相同的隐性表型，属于同一遗传互补组。此外，已知抑制金属蛋白酶的两种结构不同的药剂TAPI-2和1，10-菲咯啉阻断PMA激活的proTGF-α、细胞粘附受体L-选择素、白细胞介素6受体α亚基、β-淀粉样前体蛋白和一整套匿名中国仓鼠卵巢细胞表面蛋白的脱落。某些丝氨酸蛋白酶抑制剂通过干扰这些蛋白质的成熟和转运到细胞表面来防止这些蛋白质的释放，但不抑制胞外域从细胞表面脱落。结果表明，存在一个共同的系统膜蛋白胞外域脱落涉及一个或几个蛋白水解活性敏感的金属蛋白酶抑制剂，其作用能力可以被破坏的隐性突变在一个单一的基因。
The extracellular domains of a diverse group of membrane proteins are shed in response to protein kinase C activators such as phorbol 12-myristate 13-acetate (PMA). The lack of sequence similarity in the cleavage sites suggests the involvement-of many proteases of diverse specificity in this process. However, a mutant Chinese hamster ovary cell line recently isolated for being defective in PMA-activated shedding of the membrane-anchored growth factor transforming growth factor alpha precursor (proTGF-alpha) is concomitantly defective in the shedding of many other unrelated membrane proteins. Here we show that independent mutagenesis and selection experiments yield shedding mutants having the same recessive phenotype and belonging to the same genetic complementation group. Furthermore, two structurally distinct agents, TAPI-2 and 1,10-phenanthroline, which are known to inhibit metalloproteases, block PMA-activated shedding of proTGF-alpha, cell adhesion receptor L-selectin, interleukin 6 receptor alpha subunit, beta-amyloid precursor protein, and an entire set of anonymous Chinese hamster ovary cell surface proteins. Certain serine protease inhibitors prevent release of these proteins by interfering with their maturation and transport to the cell surface but do not inhibit ectodomain shedding hom the cell surface. The results suggest the existence of a common system for membrane protein ectodomain shedding involving one or several proteolytic activities sensitive to metalloprotease inhibitors, whose ability to act can be disrupted by recessive mutations in a single gene.