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 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.

The β1-Integrin–Dependent Function of RECK in Physiologic and Tumor Angiogenesis

“…Tumor cells were cultured in DMEM supplemented with 10% FCS. Transfection of HT1080 cells was described (16).…”

“…The product of this gene can directly bind to a series of metalloendopeptidases, including MMP-9 (9), MMP-2 (15), membrane type 1 (MT1)-MMP, CD13/ APN (16), and ADAM10 (17), and attenuate their proteolytic activity competitively in most of cases. Two metalloprotease substrate-like domains recently discovered in RECK may provide a structural basis for its biochemical actions.…”

“…Because of this structural feature, RECK controls metalloendopeptidase activities in highly limited distance from the membrane surface (perimembrane area), and moreover, it exerts unique functions by interacting with membrane-tethered metalloendopeptidases. Indeed, molecular interaction between RECK and MT1-MMP enables MT1-MMP to translocate to the lipid raft fraction, which consequently changes the route of MT1-MMP endocytosis to one that is specific to that of glycosylphosphatidylinositol-anchored proteins; this results in a shorter half-life of MT1-MMP (16). Furthermore, RECK was found to stabilize the interaction between membrane-bound Notch ligands and Notch receptors by protecting Notch ligands from ADAM10-mediated ectodomain shedding, which critically contributes to the maintenance of the self-renewal property of neural stem cells (17).…”

Involvement of the SKP2–p27KIP1 pathway in suppression of cancer cell proliferation by RECK

“…RECK was first isolated as a transformation-suppressor gene by cDNA expression cloning (Noda et al, 1989;Takahashi et al, 1998) and found to encode a membraneanchored regulator of several extracellular metalloproteinases, including MMP2, MMP7, MMP9, MT1-MMP, ADAM10 and CD13 (Takahashi et al, 1998;Oh et al, 2001;Miki et al, 2007;Muraguchi et al, 2007;Omura et al, 2009). These proteases have been implicated in tissue remodeling and cleavage of various extracellular molecules under normal and pathological conditions (Sternlicht and Werb, 2001).…”

Expression of RECK in endothelial cells of glioma: comparison with CD34 and VEGF expressions

“…Reversion-inducing cysteine-rich protein with Kazal motifs (RECK) acts as a membrane-anchored metalloproteinase regulator [1][2][3][4] which functions as a regulator of extracellular matrix integrity in normal condition, contributing to tumor and metastasis suppressing agent. However, recent study found abundant expression of RECK in the cells associated with blood vessels undergoing rapid remodeling in the mouse implantation chambers, suggesting that RECK has a role in vascular remodeling which may involve non-sprouting mechanisms such as intussusception and pruning.…”