Vitronectin's basic domain is a syndecan ligand which functions in trans to regulate vitronectin turnover.
During the process of tissue remodeling, vitronectin (Vn) is deposited in the extracellular matrix where it plays a key role in the regulation of pericellular proteolysis and cell motility. In previous studies we have shown that extracellular levels of vitronectin are controlled by receptor-mediated endocytosis and that this process is dependent upon vitronectin binding to sulfated proteoglycans. We have now identified vitronectin's 12 amino acid "basic domain" which is contained within the larger 40 amino acid heparin binding domain, as a syndecan binding site. Recombinant vitronectins representing wild type vitronectin (rVn) and vitronectin with the basic domain deleted (rVnDelta347-358) were prepared in a baculoviral expression system. The rVn as well as a glutathione S-transferase (GST) fusion protein, consisting of vitronectin's 40 amino acid heparin binding domain (GST-VnHBD), exhibited dose dependent binding to HT-1080 cell surfaces, which was attenuated following deletion of the basic domain. In addition, GST-VnHBD supported both HT-1080 and dermal fibroblast cell adhesion, which was also dependent upon the basic domain. Similarly, ARH-77 cells transfected with syndecans -1, -2, or -4, but not Glypican-1, adhered to GST-VnHBD coated wells, while adhesion of these same cells was lost following deletion of the basic domain. HT-1080 cells were unable to degrade rVnDelta347-358. Degradation of rVnDelta347-358 was completely recovered in the presence of GST-VnHBD but not in the presence of GST-VnHBDDelta347-358. These results indicate that turnover of soluble vitronectin requires ligation of vitronectin's basic domain and that this binding event can work in trans to regulate vitronectin degradation.
Wilkins-Port
Cynthia E
CE
Center for Cell Biology and Cancer Research, Neil Hellman Medical Research Building, Albany Medical College, 47 New Scotland Avenue, Albany, NY 12208, USA.
Sanderson
Ralph D
RD
Tominna-Sebald
Eiman
E
McKeown-Longo
Paula J
PJ
D000595:Amino Acid Sequence / Q000235:genetics; D001665:Binding Sites / Q000235:genetics; D002448:Cell Adhesion / Q000235:genetics*; D045744:Cell Line, Tumor; D002462:Cell Membrane / Q000378:metabolism*; D005109:Extracellular Matrix / Q000378:metabolism*; D005347:Fibroblasts / Q000378:metabolism; D005982:Glutathione Transferase / Q000235:genetics / Q000378:metabolism; D019812:Heparan Sulfate Proteoglycans / Q000235:genetics / Q000378:metabolism; D006801:Humans; D008024:Ligands; D008562:Membrane Glycoproteins / Q000235:genetics / Q000378:metabolism*; D011485:Protein Binding / Q000235:genetics*; D020033:Protein Isoforms / Q000235:genetics / Q000378:metabolism; D017434:Protein Structure, Tertiary / Q000235:genetics; D011509:Proteoglycans / Q000235:genetics / Q000378:metabolism*; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D053667:Syndecans; D014162:Transfection; D019096:Vitronectin / Q000737:chemistry / Q000378:metabolism*
D019812:Heparan Sulfate Proteoglycans; D008024:Ligands; D008562:Membrane Glycoproteins; D020033:Protein Isoforms; D011509:Proteoglycans; D011993:Recombinant Fusion Proteins; D053667:Syndecans; D019096:Vitronectin; D005982:Glutathione Transferase