title: 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.
abstract: 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.
authors.affiliation: Department of Molecular Oncology, Kyoto University Graduate School of Medicine, Kyoto 606-8501, Japan.
authors.forename: Takao
authors.initials: T
authors.lastname: Miki
authors.forename: Yujiro
authors.initials: Y
authors.lastname: Takegami
authors.forename: Katsuya
authors.initials: K
authors.lastname: Okawa
authors.forename: Teruyuki
authors.initials: T
authors.lastname: Muraguchi
authors.forename: Makoto
authors.initials: M
authors.lastname: Noda
authors.forename: Chiaki
authors.initials: C
authors.lastname: Takahashi
mesh_terms: D000626:Aminopeptidases / Q000737:chemistry; D019921:Biotinylation; D018826:CD13 Antigens / Q000378:metabolism*; D045744:Cell Line, Tumor; D002499:Centrifugation, Density Gradient; D002966:Clathrin / Q000737:chemistry; D004705:Endocytosis; D058851:GPI-Linked Proteins; D006801:Humans; D007700:Kinetics; D053511:Matrix Metalloproteinase 14 / Q000378:metabolism*; D008562:Membrane Glycoproteins / Q000737:chemistry / Q000502:physiology*; D008856:Microscopy, Fluorescence; D011994:Recombinant Proteins / Q000737:chemistry; D013395:Sucrose / Q000494:pharmacology; D014168:Transferrin / Q000737:chemistry
chemical_list: D002966:Clathrin; D058851:GPI-Linked Proteins; D008562:Membrane Glycoproteins; C115460:RECK protein, human; D011994:Recombinant Proteins; D014168:Transferrin; D013395:Sucrose; D000626:Aminopeptidases; D018826:CD13 Antigens; D053511:Matrix Metalloproteinase 14