title: Oversulfated chondroitin/dermatan sulfates containing GlcAbeta1/IdoAalpha1-3GalNAc(4,6-O-disulfate) interact with L- and P-selectin and chemokines.
abstract: We previously reported that versican, a large chondroitin/dermatan sulfate (CS/DS) proteoglycan, interacts through its CS/DS chains with adhesion molecules L- and P-selectin and CD44, as well as chemokines. Here, we have characterized these interactions further. Using a metabolic inhibitor of sulfation, sodium chlorate, we show that the interactions of the CS/DS chains of versican with L- and P-selectin and chemokines are sulfation-dependent but the interaction with CD44 is sulfation-independent. Consistently, versican's binding to L- and P-selectin and chemokines is specifically inhibited by oversulfated CS/DS chains containing GlcAbeta1-3GalNAc(4,6-O-disulfate) or IdoAalpha1-3GalNAc(4,6-O-disulfate), but its binding to CD44 is inhibited by all the CS/DS chains, including low-sulfated and unsulfated ones. Affinity and kinetic analyses using surface plasmon resonance revealed that the oversulfated CS/DS chains containing GlcAbeta1/IdoAalpha1-3GalNAc(4,6-O-disulfate) bind directly to selectins and chemokines with high affinity (K(d) 21.1 to 293 nm). In addition, a tetrasaccharide fragment of repeating GlcAbeta1-3GalNAc(4,6-O-disulfate) units directly interacts with L- and P-selectin and chemokines and oversulfated CS/DS chains containing GlcAbeta1/IdoAalpha1-3GalNAc(4,6-O-disulfate) inhibit chemokine-induced Ca(2+) mobilization. Taken together, our results show that oversulfated CS/DS chains containing GlcAbeta1/IdoAalpha1-3GalNAc(4,6-O-disulfate) are recognized by L- and P-selectin and chemokines, and imply that these chains are important in selectin- and/or chemokine-mediated cellular responses.
authors.affiliation: Laboratory of Molecular and Cellular Recognition, Osaka University Graduate School of Medicine, 2-2 Yamada-oka, Suita 565-0871, Japan. hkawashima@burnham.org
authors.forename: Hiroto
authors.initials: H
authors.lastname: Kawashima
authors.forename: Kazuyuki
authors.initials: K
authors.lastname: Atarashi
authors.forename: Mayumi
authors.initials: M
authors.lastname: Hirose
authors.forename: Jun
authors.initials: J
authors.lastname: Hirose
authors.forename: Shuhei
authors.initials: S
authors.lastname: Yamada
authors.forename: Kazuyuki
authors.initials: K
authors.lastname: Sugahara
authors.forename: Masayuki
authors.initials: M
authors.lastname: Miyasaka
mesh_terms: D001483:Base Sequence; D002809:Chondroitin Sulfates / Q000378:metabolism*; D017931:DNA Primers; D003871:Dermatan Sulfate / Q000378:metabolism*; D006801:Humans; D007700:Kinetics; D019041:L-Selectin / Q000378:metabolism*; D009844:Oligosaccharides / Q000378:metabolism*; D019007:P-Selectin / Q000378:metabolism*; D011485:Protein Binding
chemical_list: D017931:DNA Primers; D009844:Oligosaccharides; D019007:P-Selectin; D019041:L-Selectin; D003871:Dermatan Sulfate; D002809:Chondroitin Sulfates