A liquid-phase binding analysis for L-selectin. A strong dependency on highly clustered sulfate groups.

L-selectin, together with E- and P-selectins, forms a newer group of cell adhesion molecules which are believed to interact with carbohydrate ligands [Lasky, L. A., Singer, M. S., Yednoch, T. A., Dowbenko, D., Fennie, C., Rodriguez, H., Nguyen, T., Stachel, S. & Rosen, S. D. (1989) Cell 56, 1045-1055]. Using radiolabeled fucoidan as a reference ligand, we have developed a new liquid-phase microcentrifugation assay where fine differences in binding affinity can be compared accurately. We found that glucan sulfates strongly inhibited the binding of fucoidan by murine L-selectin-IgG chimera. The efficacy of inhibition is extremely dependent on the size (up to 12 kDa) and the sulfate density (up to two sulfate groups/glucose molecule) of the glucan sulfates. The nature of the inter-glucose linkages is also important. These data suggest that the binding by L-selectin prefers certain clustering and proper spatial arrangement of the anionic groups.