Identification of proteases involved in the proteolysis of vascular endothelium cadherin during neutrophil transmigration
Identification of proteases involved in the proteolysis of vascular endothelium cadherin during neutrophil transmigration
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DOI：
10.1074/jbc.m300351200
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发表时间：
2003-04-18
影响因子：
4.8
通讯作者：
Gulino-Debrac, D
中科院分区：
文献类型：
--
作者：
Hermant, B;Bibert, S;Gulino-Debrac, D
中性粒细胞跨内皮的迁移发生在表达血管内皮钙粘蛋白的细胞-细胞连接处。这种粘附受体先前被证明参与维持内皮完整性。我们建议，中性粒细胞穿越血管内皮细胞的迁移与弹性蛋白酶和组织蛋白酶G存在于中性粒细胞表面的VE钙粘蛋白的切割平行。这一假设得到以下证据的支持。1)当中性粒细胞粘附到内皮细胞单层上时，VE钙粘蛋白的蛋白水解片段被释放到培养基中; 2）在中性粒细胞粘附到内皮细胞单层上后获得的条件培养基切割重组表达的VE钙粘蛋白胞外结构域; 3）这些切割被弹性蛋白酶抑制剂抑制; 4）N-末端测序和质谱分析表明，条件培养基和外源性弹性蛋白酶产生的VE钙粘蛋白片段完全相同; 5）在与肿瘤坏死因子α刺激的和固定的嗜中性粒细胞孵育时产生弹性蛋白酶和组织蛋白酶G特异性VE钙粘蛋白切割模式; 6）在加入弹性蛋白酶或组织蛋白酶G时体外跨内皮渗透性增加;和7）在弹性蛋白酶和组织蛋白酶G抑制剂存在下，中性粒细胞的迁移在体外减少。我们的研究结果表明，中性粒细胞表面结合的蛋白酶切割VE钙粘蛋白诱导形成的间隙，通过该间隙中性粒细胞迁移。
Transmigration of neutrophils across the endothelium occurs at the cell-cell junctions where the vascular endothelium cadherin WE cadherin) is expressed. This adhesive receptor was previously demonstrated to be involved in the maintenance of endothelium integrity. We propose that neutrophil transmigration across the vascular endothelium goes in parallel with cleavage of VE cadherin by elastase and cathepsin G present on the surface of neutrophils. This hypothesis is supported by the following lines of evidence. 1) Proteolytic fragments of VE cadherin are released into the culture medium upon adhesion of neutrophils to endothelial cell monolayers; 2) conditioned culture medium, obtained after neutrophil adhesion to endothelial monolayers, cleaves the recombinantly expressed VE cadherin extracellular domain; 3) these cleavages are inhibited by inhibitors of elastase; 4) VE cadherin fragments produced by conditioned culture medium or by exogenously added elastase are identical as shown by N-terminal sequencing and mass spectrometry analysis; 5) both elastase- and cathepsin G-specific VE cadherin cleavage patterns are produced upon incubation with tumor necrosis factor a-stimulated and fixed neutrophils; 6) transendothelial permeability increases in vitro upon addition of either elastase or cathepsin G; and 7) neutrophil transmigration is reduced in vitro in the presence of elastase and cathepsin G inhibitors. Our results suggest that cleavage of VE cadherin by neutrophil surface-bound proteases induces formation of gaps through which neutrophils transmigrate.