The reversion-inducing cysteine-rich protein with Kazal motifs (RECK) interacts with membrane type 1 matrix metalloproteinase and CD13/aminopeptidase n and modulates their endocytic pathways
The reversion-inducing cysteine-rich protein with Kazal motifs (RECK) interacts with membrane type 1 matrix metalloproteinase and CD13/aminopeptidase n and modulates their endocytic pathways
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DOI：
10.1074/jbc.m610948200
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发表时间：
2007-04-20
影响因子：
4.8
通讯作者：
Takahashi, Chiaki
中科院分区：
文献类型：
--
作者：
Miki, Takao;Takegami, Yujiro;Takahashi, Chiaki
具有Kazal基序的逆转诱导富含半胱氨酸的蛋白(RECK)通过糖基磷脂酰肌醇固定在细胞表面。该分子拮抗膜型基质金属蛋白酶1(MT1-MMPs)促进原MMP2成熟的功能。在这里，我们试图阐明RECK函数背后的机制。首先，我们发现RECK与MT1-基质金属蛋白酶形成复合体，并抑制其蛋白水解性。值得注意的是，在蔗糖梯度超速离心过程中，RECK增加了与抗洗涤剂膜相关的MT1-MMP量。此外，膜胆固醇的扰动显著影响了RECK抑制MT1-MMP功能的功能。这些发现表明，RECK可能通过调节其在细胞表面的行为以及通过酶作用来调节MT1-MMPs的功能；这促使我们找到另一种分子，其在抗洗剂膜上的行为受到RECK的影响。随后，我们发现RECK与CD13/氨基肽酶N相互作用。此外，我们还发现RECK以一种胆固醇扰动敏感的方式抑制CD13的蛋白分解活性。最后，我们研究了RECK在MT1-MMP和CD13从细胞表面内化的过程中是否影响它们的行为。在没有RECK的情况下，MT1-MMPs和CD13与依赖于细胞胞吞的胞膜蛋白或小窝的标志物一起内化。然而，有趣的是，在RECK存在的情况下，这些分子优先内化，内吞标记既不依赖于笼蛋白，也不依赖于小窝，这表明RECK调节MT1-MMP和CD13的内吞途径。这种调节与MT1-MMPs和CD13的加速内化和衰变有关。本研究揭示了RECK的新功能和靶向分子。
The reversion-inducing cysteine-rich protein with Kazal motifs ( RECK) is anchored to the cell surface via glycosylphosphatidylinositol. This molecule antagonizes the function of membrane type 1 matrix metalloproteinase ( MT1-MMP) to promote proMMP-2 maturation. Here, we attempt to clarify the mechanism underlying RECK functions. First, we found that RECK forms a complex with MT1-MMP and inhibits its proteolytic activity. Notably, RECK increases the amount of MT1-MMP that associates with detergent-resistant membranes during sucrose gradient ultracentrifugation. Furthermore, perturbation of membrane cholesterol significantly affected the function of RECK in suppressing MT1-MMP function. These findings indicate that RECK possibly regulates MT1-MMP function by modulating its behavior on the cell surface as well as by enzymatic action; this prompted us to find another molecule whose behavior in detergent-resistant membranes is influenced by RECK. Subsequently, we found that RECK interacts with CD13/aminopeptidase N. Further, we found that RECK inhibits the proteolytic activity of CD13 in a cholesterol perturbation-sensitive manner. Finally, we examined whether RECK influences the behavior of MT1-MMP and CD13 during their internalization from the cell surface. In the absence of RECK, MT1-MMP and CD13 were internalized along with the markers of clathrin- or caveolae-dependent endocytosis. However, interestingly, in the presence of RECK these molecules were internalized preferentially with an endocytic marker that is neither clathrin- nor caveolae-dependent, indicating that RECK modulates endocytic pathways of MT1-MMP and CD13. This modulation was correlated with the accelerated internalization and decay of MT1-MMP and CD13. This study unveils the novel function and target molecules of RECK.