Cleavage of syndecan-1 by membrane type matrix metalloproteinase-1 stimulates cell migration
Cleavage of syndecan-1 by membrane type matrix metalloproteinase-1 stimulates cell migration
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DOI：
10.1074/jbc.m306736200
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发表时间：
2003-10-17
影响因子：
4.8
通讯作者：
Sato, H
中科院分区：
文献类型：
--
作者：
Endo, K;Takino, T;Sato, H
通过表达克隆方法从人胎盘cDNA文库中鉴定出跨膜硫酸乙酰肝素蛋白聚糖syndecan-1，它是与膜型基质金属蛋白酶-1（MT 1-MMP）相互作用的基因产物。MT 1-MMP与syndecan-1在HEK 293 T细胞中的共表达促进syndecan-1的脱落，并且细胞相关的syndecan-1的浓度降低。用MMP抑制剂BB-94或MMP的组织抑制剂（TIMP）-2而不是TIMP-1处理细胞干扰由MT 1-MMP表达促进的多配体蛋白聚糖-1脱落。相反，由12-O-十四烷酰基佛波醇-13-乙酸酯处理诱导的多配体蛋白聚糖-1脱落被BB-94抑制，但不被TIMP-1或TIMP-2抑制。多配体蛋白聚糖-1的脱落也被MT3-MMP诱导，但不被其他MT-MMP诱导。重组Syndecan-1核心蛋白被重组MT 1-MMP或MT3-MMP优先切割Gly（245）-Leu（246）肽键。用syndecan-1 cDNA稳定转染的HT 1080纤维肉瘤细胞（HT 1080/SDC）表达内源性MT 1-MMP，自发地脱落syndecan-1。HT 1080/ SDC细胞在胶原包被的培养皿上的迁移明显慢于对照HT 1080细胞。用BB-94或TIMP-2处理HT 1080/ SDC细胞可诱导syndecan-1在细胞表面积聚，同时进一步阻滞细胞迁移。用Leu取代syndecan-1的Gly（245）显著减少了HT 1080/ SDC细胞的脱落和细胞迁移。这些结果表明，MT 1-MMP通过优先切割Gly（245）-Leu（246）肽键促进syndecan-1的脱落，从而刺激细胞迁移。
The transmembrane heparan sulfate proteoglycan syndecan-1 was identified from a human placenta cDNA library by the expression cloning method as a gene product that interacts with membrane type matrix metalloproteinase-1 (MT1-MMP). Co-expression of MT1-MMP with syndecan-1 in HEK293T cells promoted syndecan-1 shedding, and concentration of cell-associated syndecan-1 was reduced. Treatment of cells with MMP inhibitor BB-94 or tissue inhibitor of MMP (TIMP)-2 but not TIMP-1 interfered with the syndecan-1 shedding promoted by MT1-MMP expression. In contrast, syndecan-1 shedding induced by 12-O-tetradecanoylphorbol-13-acetate treatment was inhibited by BB-94 but not by either TIMP-1 or TIMP-2. Shedding of syndecan-1 was also induced by MT3-MMP but not by other MT-MMPs. Recombinant syndecan-1 core protein was shown to be cleaved by recombinant MT1-MMP or MT3-MMP preferentially at the Gly(245)-Leu(246) peptide bond. HT1080 fibrosarcoma cells stably transfected with the syndecan-1 cDNA (HT1080/SDC), which express endogenous MT1-MMP, spontaneously shed syndecan-1. Migration of HT1080/ SDC cells on collagen-coated dishes was significantly slower than that of control HT1080 cells. Treatment of HT1080/ SDC cells with BB-94 or TIMP-2 induced accumulation of syndecan-1 on the cell surface, concomitant with further retardation of cell migration. Substitution of Gly(245) of syndecan-1 with Leu significantly reduced shedding from HT1080/ SDC cells and cell migration. These results suggest that the shedding of syndecan-1 promoted by MT1-MMP through the preferential cleavage of Gly(245)-Leu(246) peptide bond stimulates cell migration.