Chemically synthesized solid phase oligosaccharide probes for carbohydrate-binding receptors. Interactions of the E-, L- and P-selectins with sialyl-Le(x) and O-sulphated forms linked to biotin or to polyacrylamide

Research output: Contribution to journalArticle

Authors

  • Christine Leteux
  • Tatyana Ovchinnikova
  • Yury Tsvetkov
  • Nikolay Nifant'ev
  • Ten Feizi

Colleges, School and Institutes

Abstract

There is a growing interest in chemically defined oligosaccharide reagents for identifying proteins that bind carbohydrates and determining the specificities of carbohydrate-binding proteins. Here, we compare three sets of chemically synthesized commercially available oligosaccharide conjugates as immobilized probes, for the binding signals that they elicit with known carbohydrate-binding receptors of the immune system, the E-, P- and L-selectins. The first set of conjugates is of oligosaccharides linked to biotin via a nine-carbon spacer. The second and third sets are multivalent derivatives in which the oligosaccharides are linked, via a three-carbon spacer to poly[N-(2-hydroxyethyl)acrylamide] (PAA) or to biotinylated PAA with an average of 20% substitution of the hydroxyethyl-amide groups by carbohydrate. The conjugates were immobilized on streptavidin-coated microwells if biotinylated, otherwise by drying in uncoated wells. The most robust binding curves, overall, were with the biotinylated PAA derivatives of the ligands immobilized on streptavidin wells. These reagents have permitted a reevaluation of selectin binding signals elicited by sialyl-Lewis(x) (SLe(x)) analogues having sulphate at position 6 of the galactose (6'SuSLe(x)) or of the N-acetylglucosamine (6SuSLe(x)). The results clarify the role of 6SuSLe(x), rather then 6'SuSLe(x), as a ligand for the selectins.

Details

Original languageEnglish
Pages (from-to)53-8
Number of pages6
JournalJournal of Immunological Methods
Volume264
Issue number1-2
Publication statusPublished - 1 Jun 2002

Keywords

  • Acrylic Resins, Animals, Biotin, Carbohydrate Conformation, Carbohydrate Metabolism, Carbohydrate Sequence, E-Selectin, L-Selectin, Lewis Blood-Group System, Ligands, Molecular Probes, Molecular Sequence Data, Oligosaccharides, P-Selectin, Protein Binding, Rats, Selectins, Comparative Study, Journal Article, Research Support, Non-U.S. Gov't