title: Disintegrin interaction with alpha V beta 3 integrin on human umbilical vein endothelial cells: expression of ligand-induced binding site on beta 3 subunit.
abstract: The effect of seven disintegrins (albolabrin, barbourin, bitistatin, echistatin, eristostatin, flavoridin, and kistrin) and the neurotoxin analogue, mambin, on the adhesion of human umbilical vein endothelial cells (HUVEC) to immobilized vitronectin and fibronectin has been studied. Adhesion to vitronectin was significantly inhibited by echistatin, kistrin, flavoridin, and mambin. Echistatin, flavoridin, and kistrin bound with high affinity to immobilized alpha V beta 3 in solid phase assay; other disintegrins bound at a much lower level. Echistatin and flavoridin had a modest inhibitory effect on HUVEC adhesion to fibronectin. HUVEC adhered to disintegrins with a high selectivity toward bitistatin, echistatin, flavoridin, kistrin, and mambin. Adhesion of HUVEC to fibronectin and vitronectin resulted in cell spreading, whereas cells adhering to immobilized echistatin remained globular and cells adhering to kistrin showed abnormal morphology. Echistatin and kistrin potently inhibited the binding of monoclonal antibody (Mab) 7E3, which recognizes the alpha V beta 3 complex, to HUVEC. Echistatin and kistrin also induced the binding to HUVEC of Mab 62, which recognizes the ligand-induced binding site (LIBS) epitope on the beta 3 subunit, enhancing HUVEC binding to immobilized Mab 62. Similar results with both antibodies were obtained in Chinese hamster ovary cells transfected with alpha V beta 3 genes. In conclusion, disintegrin interaction with HUVEC appears to be selectively mediated by alpha V beta 3 receptors, and it results in an expression of LIBS epitope that may play a role in the regulation of ligand-binding affinity and intracellular signaling.
authors.affiliation: Department of Physiology, Temple University School of Medicine, Phildelphia, PA 19140, USA.
authors.forename: D
authors.initials: D
authors.lastname: Juliano
authors.forename: Y
authors.initials: Y
authors.lastname: Wang
authors.forename: C
authors.initials: C
authors.lastname: Marcinkiewicz
authors.forename: L A
authors.initials: LA
authors.lastname: Rosenthal
authors.forename: G J
authors.initials: GJ
authors.lastname: Stewart
authors.forename: S
authors.initials: S
authors.lastname: Niewiarowski
mesh_terms: D000818:Animals; D016466:CHO Cells; D002448:Cell Adhesion / Q000502:physiology*; D006224:Cricetinae; D019483:Disintegrins; D004730:Endothelium, Vascular / Q000166:cytology* / Q000276:immunology; D005353:Fibronectins / Q000502:physiology*; D006801:Humans; D008024:Ligands; D010455:Peptides / Q000276:immunology / Q000502:physiology*; D019030:Receptors, Vitronectin / Q000502:physiology*; D011994:Recombinant Proteins; D014471:Umbilical Veins; D017819:Viperidae; D019096:Vitronectin / Q000502:physiology*
chemical_list: D019483:Disintegrins; D005353:Fibronectins; D008024:Ligands; D010455:Peptides; D019030:Receptors, Vitronectin; D011994:Recombinant Proteins; D019096:Vitronectin