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.
Yoshida
T
T
Department of Biology, Johns Hopkins University, Baltimore, MD 21218.
Fennie
C
C
Lasky
L A
LA
Lee
Y C
YC
D000925:Anticoagulants / Q000378:metabolism; D002240:Carbohydrate Sequence; D015815:Cell Adhesion Molecules / Q000737:chemistry* / Q000378:metabolism*; D006801:Humans; D007074:Immunoglobulin G / Q000737:chemistry / Q000378:metabolism; D007700:Kinetics; D019041:L-Selectin; D008562:Membrane Glycoproteins / Q000737:chemistry / Q000378:metabolism; D008969:Molecular Sequence Data; D011134:Polysaccharides / Q000737:chemistry / Q000378:metabolism / Q000494:pharmacology*; D011485:Protein Binding; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000378:metabolism; D013329:Structure-Activity Relationship; D013379:Substrate Specificity
D000925:Anticoagulants; D015815:Cell Adhesion Molecules; D007074:Immunoglobulin G; D008562:Membrane Glycoproteins; D011134:Polysaccharides; D011993:Recombinant Fusion Proteins; D019041:L-Selectin; C007789:fucoidan