A Selective Extracellular Matrix Proteomics Approach Identifies Fibronectin Proteolysis by A Disintegrin-like and Metalloprotease Domain with Thrombospondin Type 1 Motifs (ADAMTS16) and Its Impact on Spheroid Morphogenesis.
A Selective Extracellular Matrix Proteomics Approach Identifies Fibronectin Proteolysis by A Disintegrin-like and Metalloprotease Domain with Thrombospondin Type 1 Motifs (ADAMTS16) and Its Impact on Spheroid Morphogenesis.
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选择性细胞外基质蛋白质组学方法通过具有血小板反应蛋白 1 型基序 (ADAMTS16) 的解整合素样和金属蛋白酶结构域 (ADAMTS16) 鉴定纤连蛋白的蛋白水解作用及其对球体形态发生的影响。
DOI：
10.1074/mcp.ra118.000676
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发表时间：
2018
期刊：
影响因子：
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通讯作者：
Chiquet-Ehrismann,Ruth
中科院分区：
文献类型：
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作者：
Schnellmann,Rahel;Sack,Ragna;Hess,Daniel;Annis,DouglasS;Mosher,DeaneF;Apte,SuneelS;Chiquet-Ehrismann,Ruth
分泌蛋白酶和细胞表面蛋白酶是细胞外基质（ECM）转化的主要介质，但其机制和调控作用尚不清楚。我们开发了一种质谱方法，使用体外生产的无细胞ECM来鉴定纤维连接蛋白（FN）作为分泌金属蛋白酶ADAMTS16的新底物。ADAMTS16在其(I)5和(I)6模块之间切割FN，释放FN自组装必需的n端30 kDa肝素结合结构域。ADAMTS16损害FN纤维形成以及纤维蛋白1和腱蛋白c的组装，从而抑制培养成纤维细胞形成成熟的ECM。此外，ADAMTS16对肾小管来源的MDCKI细胞的球体形成具有显著的形态发生影响。ADAMTS16释放的n端FN结构域上调MMP3，使FN的(I)5-(I)6连接子与ADAMTS16相似，从而形成蛋白水解前馈机制。因此，FN蛋白水解不仅调节FN周转，还调节FN组装，对ECM组装和形态发生具有潜在的长期影响。
Secreted and cell-surface proteases are major mediators of extracellular matrix (ECM) turnover, but their mechanisms and regulatory impact are poorly understood. We developed a mass spectrometry approach using a cell-free ECM producedin vitroto identify fibronectin (FN) as a novel substrate of the secreted metalloprotease ADAMTS16. ADAMTS16 cleaves FN between its (I)5and (I)6modules, releasing the N-terminal 30 kDa heparin-binding domain essential for FN self-assembly. ADAMTS16 impairs FN fibrillogenesis as well as fibrillin-1 and tenascin-C assembly, thus inhibiting formation of a mature ECM by cultured fibroblasts. Furthermore ADAMTS16 has a marked morphogenetic impact on spheroid formation by renal tubule-derived MDCKI cells. The N-terminal FN domain released by ADAMTS16 up-regulates MMP3, which cleaves the (I)5-(I)6linker of FN similar to ADAMTS16, therefore creating a proteolytic feed-forward mechanism. Thus, FN proteolysis not only regulates FN turnover, but also FN assembly, with potential long-term consequences for ECM assembly and morphogenesis.