Shedding of Discoidin Domain Receptor 1 by Membrane-type Matrix Metalloproteinases

• Published in: The Journal of biological chemistry 2013-04, Vol.288 (17), p.12114-12129
• Main Authors: Fu, Hsueh-Liang, Sohail, Anjum, Valiathan, Rajeshwari R., Wasinski, Benjamin D., Kumarasiri, Malika, Mahasenan, Kiran V., Bernardo, M.Margarida, Tokmina-Roszyk, Dorota, Fields, Gregg B., Mobashery, Shahriar, Fridman, Rafael
• Format: Article
• Language: English
• Subjects: Amino Acid Motifs; Animals; Cell Line, Tumor; Cell Surface Receptor; Chlorocebus aethiops; Collagen; Collagenases - genetics; Collagenases - metabolism; COS Cells; Discoidin Domain Receptor 1; Discoidin Domain Receptors; Enzymology; Extracellular Matrix - genetics; Extracellular Matrix - metabolism; Female; Humans; Matrix Metalloproteinase (MMP); Protein Structure, Tertiary; Proteolysis; Proteolytic Enzymes; Receptor Protein-Tyrosine Kinases - genetics; Receptor Protein-Tyrosine Kinases - metabolism; Shedding
• ISSN: 0021-9258; 1083-351X
• DOI: 10.1074/jbc.M112.409599

The discoidin domain receptors (DDRs) are receptor tyrosine kinases that upon binding to collagens undergo receptor phosphorylation, which in turn activates signal transduction pathways that regulate cell-collagen interactions. We report here that collagen-dependent DDR1 activation is partly regulated by the proteolytic activity of the membrane-anchored collagenases, MT1-, MT2-, and MT3-matrix metalloproteinase (MMP). These collagenases cleave DDR1 and attenuate collagen I- and IV-induced receptor phosphorylation. This effect is not due to ligand degradation, as it proceeds even when the receptor is stimulated with collagenase-resistant collagen I (r/r) or with a triple-helical peptide harboring the DDR recognition motif in collagens. Moreover, the secreted collagenases MMP-1 and MMP-13 and the glycosylphosphatidylinositol-anchored membrane-type MMPs (MT4- and MT6-MMP) have no effect on DDR1 cleavage or activation. N-terminal sequencing of the MT1-MMP-mediated cleaved products and mutational analyses show that cleavage of DDR1 takes place within the extracellular juxtamembrane region, generating a membrane-anchored C-terminal fragment. Metalloproteinase inhibitor studies show that constitutive shedding of endogenous DDR1 in breast cancer HCC1806 cells is partly mediated by MT1-MMP, which also regulates collagen-induced receptor activation. Taken together, these data suggest a role for the collagenase of membrane-type MMPs in regulation of DDR1 cleavage and activation at the cell-matrix interface. Background: DDR1 is a receptor tyrosine kinase that signals in response to collagen and regulates cell-collagen interactions. MT-MMPs are membrane-anchored proteases that accomplish pericellular collagenolysis. Results: MT-MMPs cleave DDR1 and regulate collagen-induced receptor phosphorylation. Conclusion: MT-MMPs negatively regulate DDR1 activation by promoting receptor ectodomain shedding. Significance: Cross-talk between membrane-anchored collagenases and RTKs integrates collagen-induced signaling and pericellular proteolysis.