Specific binding of angiogenin to calf pulmonary artery endothelial cells.
Specific binding of angiogenin (ANG) to calf pulmonary artery endothelial cells was demonstrated. Cellular binding at 4 degrees C of 125I-labeled human recombinant ANG was time and concentration dependent, reversible, and saturable in the presence of increasing amounts of the unlabeled molecules. The interaction was shown to be specific since a large excess of unlabeled ANG reduced labeled ANG binding by 80%, whereas similar doses of RNase A, a structurally related protein, had no effect. Scatchard analyses of binding data revealed two apparent components. High-affinity sites with an apparent dissociation constant of 5 x 10(-9) M were shown to represent cell-specific interactions. The second component, comprising low-affinity/high-capacity sites with an apparent dissociation constant of 0.2 x 10(-6) M, was essentially associated with pericellular components. High-affinity ANG binding sites varied with cell density and were found on other endothelial cells from bovine aorta, cornea, and adrenal cortex capillary but not on Chinese hamster lung fibroblasts. Divalent copper, a modulator of angiogenesis, was found to induce a severalfold increase in specific cell-bound radioactivity. Placental ribonuclease inhibitor, a tight-binding inhibitor of both ribonucleolytic and angiogenic activities of ANG, abolished 125I-labeled human recombinant ANG binding only in the absence of copper.
Badet
J
J
Laboratoire de Biotechnologie des Cellules Eucaryotes, Université Paris XII, Créteil, France.
Soncin
F
F
Guitton
J D
JD
Lamare
O
O
Cartwright
T
T
Barritault
D
D
D043925:Angiogenesis Inducing Agents / Q000378:metabolism*; D000818:Animals; D001667:Binding, Competitive; D002413:Cations, Divalent; D002417:Cattle; D002478:Cells, Cultured; D003300:Copper / Q000494:pharmacology; D019327:Copper Sulfate; D004730:Endothelium, Vascular / Q000378:metabolism*; D006133:Growth Substances / Q000378:metabolism*; D007700:Kinetics; D011485:Protein Binding; D011506:Proteins / Q000378:metabolism*; D011651:Pulmonary Artery; D011994:Recombinant Proteins / Q000378:metabolism; D012259:Ribonuclease, Pancreatic / Q000378:metabolism
D043925:Angiogenesis Inducing Agents; D002413:Cations, Divalent; D006133:Growth Substances; D011506:Proteins; D011994:Recombinant Proteins; D003300:Copper; C046366:angiogenin; D012259:Ribonuclease, Pancreatic; D019327:Copper Sulfate