Equilibrium Binding Analysis of Neural Cell Adhesion Molecule Binding to Heparin

Abstract

The kinetics of neural cell adhesion molecule (NCAM) binding to heparin were studied in a heparin‐Sepharose‐based solid‐phase binding assay. The observed binding is time dependent and saturable. A binding constant of 5.2 ± 1.4 × 10⁻⁸M is observed for binding of newborn rat NCAM to heparin. This is ∼25 times lower than the binding constant determined for newborn rat NCAM homophilic binding. Both Scatchard and Hill plot analyses suggest the presence of only one binding site. Fab' fragments of antibodies to rat NCAM significantly inhibit binding, a result indicating that a specific site on NCAM is involved in binding to heparin. The binding is inhibited by heparin (IC₅₀, ∼5 μg/ml), whereas chondroitin sulfate is a less potent inhibitor (IC₅₀, ∼15 μg/ml).