Pericellular activation of proMMP-7 (promatrilysin-1) through interaction with CD151
Pericellular activation of proMMP-7 (promatrilysin-1) through interaction with CD151
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DOI：
10.1038/labinvest.3700351
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发表时间：
2005-12-01
影响因子：
5
通讯作者：
Okada, Y
中科院分区：
文献类型：
--
作者：
Shiomi, T;Inoki, I;Okada, Y
基质金属蛋白酶-7（MMP-7）（也称为基质溶解素-1）作为酶原（proMMP-7）分泌，并且在活化后的各种细胞外基质（ECM）和非ECM分子的降解中起关键作用。为筛选与proMMP-7活化相关的结合蛋白，以proMMP-7为诱饵，利用酵母双杂交系统筛选人肺cDNA文库。我们鉴定了一个候选分子CD 151，它是跨膜4超家族的成员。通过来自CaR-1细胞（一种表达proMMP-7和CD 151的人直肠癌细胞系）和与proMMP-7孵育的CD 151稳定转染子的分子的免疫沉淀来证明proMMP-7与CD 151的复合物形成。使用proMMP-7和CD 151的缺失突变体的酵母双杂交测定表明proMMP-7的前肽和CD 151的COOH末端细胞外环之间的相互作用。用CD 151稳定转染子显示I-125标记的proMMP-7与细胞膜上的CD 151的结合活性。激光扫描共聚焦显微镜显示proMMP-7与CD 151共定位于CD 151稳定转染细胞和CaR-1细胞的细胞膜上。原位酶谱使用交联羧甲基化转铁蛋白，基质金属蛋白酶-7的底物，表现出蛋白酶的活性和周围的CD 151稳定的转染子和CaR-1细胞，而活性被废除的治疗与MMP抑制剂，抗MMP-7抗体或抗CD 151抗体。在人肺腺癌组织中，MMP-7和CD 151共定位于癌细胞上。金属蛋白酶活性存在于这些组织中，并可以抑制MMP-7或CD 151的抗体。这些数据首次证明了proMMP-7通过与CD 151相互作用在细胞膜上被捕获和活化，并表明类似于T1-MMP/MMP-2系统，MMP-7参与由CD 151介导的细胞周活化机制的可能性，这是在包括癌症侵袭和转移在内的各种病理生理条件下细胞膜上蛋白水解的关键步骤。
Matrix metalloproteinase-7 (MMP-7) ( also known as matrilysin-1) is secreted as a proenzyme (proMMP-7) and plays a key role in the degradation of various extracellular matrix (ECM) and non-ECM molecules after activation. To identify the binding proteins related to proMMP-7 activation, a human lung cDNA library was screened by yeast two-hybrid system using proMMP-7 as bait. We identified a candidate molecule CD151, which is a member of the transmembrane 4 superfamily. Complex formation of proMMP-7 with CD151 was demonstrated by immunoprecipitation of the molecules from CaR-1 cells, a human rectal carcinoma cell line, expressing both proMMP-7 and CD151, and CD151 stable transfectants incubated with proMMP-7. Yeast two-hybrid assays using deletion mutants of proMMP-7 and CD151 suggested an interaction between the propeptide of proMMP-7 and the COOH-terminal extracellular loop of CD151. The binding activity of I-125-labeled proMMP-7 to CD151 on the cell membranes was shown with CD151 stable transfectants. Laser-scanning confocal microscopy demonstrated that proMMP-7 colocalizes with CD151 on the cell membranes of CD151 stable transfectants and CaR-1 cells. In situ zymography using crosslinked carboxymethylated transferrin, a substrate of MMP-7, demonstrated proteinase activity on and around CD151 stable transfectants and CaR-1 cells, while the activity was abolished by their treatment with MMP inhibitors, anti-MMP-7 antibody or anti-CD151 antibody. In human lung adenocarcinoma tissues, colocalization of MMP-7 and CD151 was demonstrated on the carcinoma cells. Metalloproteinase activity was present in these tissues and could be inhibited by antibodies to MMP-7 or CD151. These data demonstrate for the first time that proMMP-7 is captured and activated on the cell membranes through interaction with CD151, and suggest the possibility that similar to the T1-MMP/MMP-2 system, MMP-7 is involved in the pericellular activation mechanism mediated by CD151, a crucial step in proteolysis on the cell membranes under various pathophysiological conditions including cancer invasion and metastasis.