Structural requirements of heparan sulfate for the binding to the tumor-derived adhesion factor/angiomodulin that induces cord-like structures to ECV-304 human carcinoma cells.
Tumor-derived adhesion factor/angiomodulin (AGM) is accumulated in tumor blood vessels and on the endothelial cell surface (Akaogi, K., Okabe, Y., Sato, J., Nagashima, Y., Yasumitsu, H., Sugahara, K., and Miyazaki, K. (1996) Proc. Natl. Acad. Sci. U. S. A. 93, 8384-8389). In cell culture, it promotes cell adhesion and morphological changes to form cord-like structures of the human bladder carcinoma cell line ECV-304. The cord formation is prevented by heparin, which inhibits the binding of AGM to ECV-304 cells. This observation suggests that AGM interacts with cell surface heparan sulfate (HS) proteoglycans. In this study, HS glycosaminoglycans and core proteins of integral transmembrane proteoglycans, syndecan-1 and -4, were identified by immunocytochemistry on ECV-304 cells, and the structural requirements for the interaction of HS with AGM were characterized. Inhibition experiments with sulfated polysaccharides and chemically modified heparin derivatives indicated that sulfate groups were essential for both AGM-HS binding and cord-like structure formation and that the rank order of the different sulfate groups in terms of their contribution was N-sulfate > 6-O-sulfate > 2-O-sulfate. The minimum size of heparin, a chemical analog of HS, required for the binding to AGM was a dodecasaccharide as determined by competition experiments using size-defined heparin oligosaccharides. Thus, a specific sulfation pattern in the HS of cell surface syndecans of ECV-304 cells is required for AGM binding and the morphological changes.
Kishibe
J
J
Department of Biochemistry, Kobe Pharmaceutical University, Higashinada-ku, Kobe 658-8558, Japan.
Yamada
S
S
Okada
Y
Y
Sato
J
J
Ito
A
A
Miyazaki
K
K
Sugahara
K
K
D000818:Animals; D002417:Cattle; D002448:Cell Adhesion / Q000502:physiology*; D002462:Cell Membrane / Q000187:drug effects / Q000378:metabolism / Q000473:pathology*; D003094:Collagen / Q000494:pharmacology; D004187:Disaccharides / Q000032:analysis / Q000737:chemistry; D006025:Glycosaminoglycans / Q000032:analysis / Q000378:metabolism*; D006497:Heparan Sulfate / Q000737:chemistry* / Q000378:metabolism*; D006801:Humans; D007150:Immunohistochemistry; D007700:Kinetics; D008099:Liver / Q000378:metabolism; D008562:Membrane Glycoproteins / Q000032:analysis / Q000378:metabolism; D009363:Neoplasm Proteins / Q000378:metabolism*; D009844:Oligosaccharides / Q000494:pharmacology; D011509:Proteoglycans / Q000032:analysis / Q000378:metabolism; D053668:Syndecan-1; D053671:Syndecan-4; D053667:Syndecans; D014407:Tumor Cells, Cultured; D001749:Urinary Bladder Neoplasms / Q000473:pathology*
D003094:Collagen; D004187:Disaccharides; D006025:Glycosaminoglycans; D006497:Heparan Sulfate; D008562:Membrane Glycoproteins; D009363:Neoplasm Proteins; D009844:Oligosaccharides; D011509:Proteoglycans; D053668:Syndecan-1; D053671:Syndecan-4; D053667:Syndecans; C508241:SDC1 protein, human; C508304:SDC4 protein, human; C086009:tumor-derived adhesion factor