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A liquid-phase binding analysis for L-selectin. A strong dependency on highly clustered sulfate groups
T Yoshida1, C Fennie, L A Lasky
1Department of Biology, Johns Hopkins University, Baltimore, MD 21218.
European Journal of Biochemistry
|June 1, 1994
Summary
Glucan sulfates strongly inhibit L-selectin binding to carbohydrate ligands, with efficacy depending on size, sulfate density, and linkage type. This suggests L-selectin prefers specific arrangements of anionic groups for binding.
Area of Science:
- Immunology
- Glycobiology
- Biochemistry
Background:
- L-selectin is a cell adhesion molecule involved in immune responses.
- Selectins are known to interact with carbohydrate ligands.
- Understanding these interactions is crucial for developing targeted therapies.
Purpose of the Study:
- To develop a novel assay for comparing binding affinities of L-selectin ligands.
- To investigate the inhibitory effects of glucan sulfates on L-selectin binding.
- To determine the structural requirements for glucan sulfates to inhibit L-selectin-ligand interactions.
Main Methods:
- Development of a liquid-phase microcentrifugation assay.
- Utilized radiolabeled fucoidan as a reference ligand.
- Tested various glucan sulfates with defined sizes and sulfate densities.
Main Results:
- Glucan sulfates effectively inhibited the binding of fucoidan to L-selectin-IgG chimera.
- Inhibition efficacy was highly dependent on glucan sulfate size (up to 12 kDa) and sulfate density (up to 2 sulfates/glucose).
- The type of inter-glucose linkages in glucan sulfates also influenced binding inhibition.
Conclusions:
- L-selectin binding to carbohydrate ligands is sensitive to the structural features of sulfated polysaccharides.
- Optimal binding involves specific clustering and spatial arrangement of anionic groups.
- This study provides insights into the molecular basis of L-selectin-mediated adhesion.