title: Valency dependent patterns of binding of human L-selectin toward sialyl and sulfated oligosaccharides of Le(a) and Le(x) types: relevance to anti-adhesion therapeutics.
abstract: The human L-selectin is known to bind to immobilized 3'-sialyl-Le(x) and -Le(a) oligosaccharides both under static and physiological flow conditions. Here the reactivities toward 3'-sulfated and 3'-sialyl-Le(a) and -Le(x) pentasaccharides are compared by in-vitro binding and inhibition assays using preparations of human L-selectin-IgG-Fc chimera in which the selectin is predominantly in di- and tetrameric form (paucivalent) or in the form of a complex with anti-IgG (multivalent). Affinity for the sulfated ligands is marginally greater than for the sialyl ligands, as judged by concentrations required to give 50% inhibition of the multivalent selectin binding to the immobilized sulfated and sialyl ligands. There is a striking difference, however, in the avidities of binding of the two L-selectin forms toward the sulfated and sialyl ligands when these are immobilized in the clustered state: the paucivalent selectin gives detectable binding only to the sulfated ligands when these are immobilized as neoglycolipids on plastic microwells (up to 100 pmol immobilized per well) whereas the multivalent L-selectin binds well to both classes of ligand. Moreover, binding of the paucivalent selectin form is effectively inhibited only by the sulfated ligand, although binding of the multivalent selectin is inhibitable by both the sulfated and sialyl ligands. Such striking valency-dependent differences in ligand binding avidity and inhibitability may be manifest in vivo with the membrane-bound L-selectin, as marked variations occur in its density of expression on leukocytes. Thus, for the purpose of selecting inhibitors for development of therapeutic anti-inflammatory compounds, experimental designs based on the paucivalent L-selectin would more clearly single out compounds with broad spectrum anti-adhesive activities toward the both the high- and low-avidity interactions of the cell adhesion protein.
authors.affiliation: The Glycosciences Laboratory, Imperial College School of Medicine, Northwick Park Hospital, Harrow, Middlesex, U.K.
authors.forename: C
authors.initials: C
authors.lastname: Galustian
authors.forename: R A
authors.initials: RA
authors.lastname: Childs
authors.forename: C T
authors.initials: CT
authors.lastname: Yuen
authors.forename: A
authors.initials: A
authors.lastname: Hasegawa
authors.forename: M
authors.initials: M
authors.lastname: Kiso
authors.forename: A
authors.initials: A
authors.lastname: Lubineau
authors.forename: G
authors.initials: G
authors.lastname: Shaw
authors.forename: T
authors.initials: T
authors.lastname: Feizi
mesh_terms: D000818:Animals; D015295:Antigens, Tumor-Associated, Carbohydrate / Q000378:metabolism*; D014408:Biomarkers, Tumor / Q000378:metabolism*; D018395:CA-19-9 Antigen; D002850:Chromatography, Gel; D002851:Chromatography, High Pressure Liquid; D004563:Electrochemistry; D005732:Gangliosides / Q000737:chemistry / Q000378:metabolism*; D006801:Humans; D019041:L-Selectin / Q000737:chemistry / Q000378:metabolism*; D007983:Lewis Blood Group Antigens / Q000378:metabolism*; D051381:Rats; D000080506:Sialyl Lewis X Antigen
chemical_list: D015295:Antigens, Tumor-Associated, Carbohydrate; D014408:Biomarkers, Tumor; D018395:CA-19-9 Antigen; D005732:Gangliosides; D007983:Lewis Blood Group Antigens; D000080506:Sialyl Lewis X Antigen; C094688:sialyl Lewis(x) ganglioside; D019041:L-Selectin; C064564:sialyl Le(a) ganglioside