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

• Published in: The Journal of biological chemistry Vol. 282; no. 16; pp. 12341 - 12352
• Main Authors: Miki, Takao, Takegami, Yujiro, Okawa, Katsuya, Muraguchi, Teruyuki, Noda, Makoto, Takahashi, Chiaki
• Format: Journal Article
• Language: English
• Published: United States Elsevier Inc 20.04.2007; American Society for Biochemistry and Molecular Biology
• Subjects: Aminopeptidases - chemistry; Biotinylation; CD13 Antigens - metabolism; Cell Line, Tumor; Centrifugation, Density Gradient; Clathrin - chemistry; Endocytosis; GPI-Linked Proteins; Humans; Kinetics; Matrix Metalloproteinase 14 - metabolism; Membrane Glycoproteins - chemistry; Membrane Glycoproteins - physiology; Microscopy, Fluorescence; Recombinant Proteins - chemistry; Sucrose - pharmacology; Transferrin - chemistry
• ISSN: 0021-9258; 1083-351X
• DOI: 10.1074/jbc.M610948200

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 clathrinnor 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.