RECK-mediated suppression of tumor cell invasion is regulated by glycosylation in human tumor cell lines.
RECK, a glycosylphosphatidylinositol (GPI)-anchored glycoprotein, negatively regulates matrix metalloproteinases (MMP), such as MMP-9, and inhibits tumor invasion and metastasis. The predicted amino acid sequence of human RECK includes five putative N-glycosylation sites; however, the precise biochemical role of glycosylated RECK remains unknown. In this study, we examined the link between glycosylation and the function of RECK in human tumor cell lines. RECK protein was glycosylated at Asn86, Asn200, Asn297, and Asn352 residues but not at the Asn39 residue in HT1080 cells. Although the glycosylation of these asparagine sites did not play a role in the cell surface localization of RECK as a GPI-anchored protein, the glycosylation of RECK Asn297 residue was involved in the suppression of MMP-9 secretion and Asn352 residue was necessary to inhibit MMP-2 activation. Moreover, RECK-suppressed tumor cell invasion was reversed by inhibiting glycosylation at Asn86, Asn297, and Asn352 residues of RECK. Thus, these findings indicate that glycosylation mediates RECK suppression of tumor cell invasion by multiple mechanisms such as suppressing MMP-9 secretion and inhibiting MMP-2 activation.
Simizu
Siro
S
Antibiotics Laboratory, Discovery Research Institute, RIKEN and Graduate School of Science and Engineering, Saitama University, Japan.
Takagi
Satoshi
S
Tamura
Yuki
Y
Osada
Hiroyuki
H
D001216:Asparagine / Q000378:metabolism; D045744:Cell Line, Tumor; D002462:Cell Membrane / Q000378:metabolism; D042783:Endothelial Cells / Q000378:metabolism; D004789:Enzyme Activation; D058851:GPI-Linked Proteins; D006031:Glycosylation; D017261:Glycosylphosphatidylinositols / Q000096:biosynthesis / Q000378:metabolism; D006801:Humans; D020778:Matrix Metalloproteinase 2 / Q000378:metabolism; D020780:Matrix Metalloproteinase 9 / Q000378:metabolism; D061965:Matrix Metalloproteinase Inhibitors; D008562:Membrane Glycoproteins / Q000096:biosynthesis / Q000378:metabolism*; D009361:Neoplasm Invasiveness
D058851:GPI-Linked Proteins; D017261:Glycosylphosphatidylinositols; D061965:Matrix Metalloproteinase Inhibitors; D008562:Membrane Glycoproteins; C115460:RECK protein, human; D001216:Asparagine; D020778:Matrix Metalloproteinase 2; D020780:Matrix Metalloproteinase 9