High and Low Affinity Binding Sites for Basic Fibroblast Growth Factor on Cultured Cells: Absence of a Role for Low Affinity Binding in the Stimulation of Plasminogen Activator Production by Bovine Capillary Endothelial Cells

J Cell Physiol. 1987 Apr;131(1):123-30. doi: 10.1002/jcp.1041310118.

Abstract

Scatchard analysis of binding of 125I-basic fibroblast growth factor (FGF) to baby hamster kidney (BHK) cells revealed the presence of two binding sites: a high affinity site with KD of 20 pM and 80,000 sites per cell and a low affinity site with KD of about 2 nM and 600,000 sites per cell. The binding to the two sites could be separated by first washing the cells with 2 M NaCl at pH 7.5 which released the low affinity binding and then extracting the cells with 0.5% Triton X-100 to recover the 125I-basic FGF bound to high affinity sites. The binding to the high affinity site was acid sensitive, suggesting that it represented binding to the receptor. Binding to the low affinity site could be competed strongly by heparin and less strongly by heparan sulfate but not by chondroitin sulfate, dermatan sulfate, or keratan sulfate. Treatment of BHK cells with heparinase abolished 62% of the low affinity binding, suggesting that the low affinity binding represented binding to cell-associated, heparin-like molecules. A variety of other cell types, including bovine capillary endothelial (BCE) cells, also demonstrated both low and high affinity binding sites. To test whether the low affinity binding might play a role in the basic FGF stimulation of plasminogen activator (PA) production by BCE cells, heparin was added to BCE cultures at concentrations which totally blocked binding of 125I-basic FGF to the low affinity sites. Addition of the heparin did not diminish the increased PA production induced by basic FGF. This suggests that the low affinity binding has no direct role in the stimulation of PA production in BCE cells.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Animals
  • Binding Sites
  • Binding, Competitive
  • Capillaries / cytology
  • Capillaries / metabolism
  • Capillaries / ultrastructure
  • Cattle
  • Cell Line
  • Cells, Cultured
  • Cricetinae
  • Endothelium / cytology
  • Endothelium / metabolism*
  • Endothelium / ultrastructure
  • Fibroblast Growth Factors / metabolism*
  • Heparin / physiology
  • Humans
  • Iodine Radioisotopes
  • Kidney / cytology
  • Kidney / metabolism
  • Kidney / ultrastructure
  • Plasminogen Activators / biosynthesis*
  • Receptors, Cell Surface / physiology
  • Receptors, Fibroblast Growth Factor

Substances

  • Iodine Radioisotopes
  • Receptors, Cell Surface
  • Receptors, Fibroblast Growth Factor
  • Fibroblast Growth Factors
  • Heparin
  • Plasminogen Activators