Investigation of the role of Endo180/urokinase-type plasminogen activator receptor-associated protein as a collagenase 3 (matrix metalloproteinase 13) receptor.
Procollagenase 3 can be activated by interaction with and cleavage by the cell-associated membrane type 1 metalloproteinase (MT1 MMP; MMP 14). It has also been shown to bind to a specific receptor, and is subsequently internalized via the low-density lipoprotein-related receptor by osteoblast cell lines. The receptor was identified as a recycling glycoprotein of the macrophage mannose receptor family, Endo180. In order to ascertain whether there is a relationship between Endo180 binding and procollagenase 3 activation, we have compared procollagenase 3 activation by an HT1080 fibrosarcoma cell line overexpressing MT1 MMP, without and with overexpression of Endo180. No difference in procollagenase 3 activation was observed, and neither was the enzyme bound to the cells or internalized. In contrast, the osteoblast cell lines, MG63 and UMR-106, both bound and internalized procollagenase 3. However, immunolocalization studies showed that the Endo180 abundantly expressed by these cells did not co-localize with the procollagenase 3. In further biochemical studies we confirmed that procollagenase 3 did not bind to Endo180, using both ligand- blotting and immunoprecipitation techniques. We conclude that Endo180 is unlikely to be a receptor for collagenase 3 in relation to either its activation or cell binding and internalization, and that other interaction partners must be sought.
Bailey
Louise
L
School of Biological Sciences, University of East Anglia, Norwich NR4 7TJ, UK. louise.bailey@uea.ac.uk
Wienke
Dirk
D
Howard
Matthew
M
Knäuper
Vera
V
Isacke
Clare M
CM
Murphy
Gillian
G
D015153:Blotting, Western; D002460:Cell Line; D017364:Collagenases / Q000378:metabolism*; D018076:DNA, Complementary / Q000378:metabolism; D006801:Humans; D008024:Ligands; D053509:Matrix Metalloproteinase 13; D008856:Microscopy, Fluorescence; D010006:Osteoblasts / Q000378:metabolism; D011233:Precipitin Tests; D011485:Protein Binding; D012333:RNA, Messenger / Q000378:metabolism; D011956:Receptors, Cell Surface / Q000737:chemistry / Q000378:metabolism*; D011975:Receptors, Mitogen / Q000737:chemistry / Q000378:metabolism*; D055293:Receptors, Urokinase Plasminogen Activator; D013997:Time Factors; D014162:Transfection; D014407:Tumor Cells, Cultured
D018076:DNA, Complementary; C408294:Endo180; D008024:Ligands; C524898:PLAUR protein, human; D012333:RNA, Messenger; D011956:Receptors, Cell Surface; D011975:Receptors, Mitogen; D055293:Receptors, Urokinase Plasminogen Activator; D017364:Collagenases; C507079:MMP13 protein, human; D053509:Matrix Metalloproteinase 13