Affinity chromatography of sulphated polysaccharides separately fractionated on antithrombin III and heparin cofactor II immobilized on concanavalin A--sepharose.
Three sulphated polysaccharides, dermatan sulphate, fucan and heparin, were fractionated according to their affinity towards antithrombin III (ATIII) and heparin cofactor II (HCII), the two main physiological thrombin (IIa) inhibitors. Both inhibitors were immobilized on concanavalin A-Sepharose, which binds to the glycosylated chains of the proteins while the protein-binding site for the polysaccharide remains free. Each polysaccharide was fractionated into bound and unbound fractions either for ATIII or HCII. The eluted fractions were tested for their ability to catalyse ATIII/IIa and HCII/IIa interactions. The possible presence of a unique binding site for ATIII and HCII, on each sulphated polysaccharide, was also studied.
Sinniger
V
V
Laboratoire d'Hématologie, Hospital Necker Enfants Malades, Paris, France.
Tapon-Bretaudière
J
J
Millien
C
C
Muller
D
D
Jozefonvicz
J
J
Fischer
A M
AM
D000818:Animals; D000990:Antithrombin III / Q000737:chemistry*; D002417:Cattle; D002846:Chromatography, Affinity; D003124:Colorimetry; D003208:Concanavalin A; D003871:Dermatan Sulfate / Q000032:analysis*; D006493:Heparin / Q000032:analysis*; D015844:Heparin Cofactor II / Q000737:chemistry*; D007413:Intestinal Mucosa / Q000737:chemistry; D011134:Polysaccharides / Q000032:analysis*; D012685:Sepharose
D011134:Polysaccharides; D003208:Concanavalin A; D003871:Dermatan Sulfate; D015844:Heparin Cofactor II; D000990:Antithrombin III; D006493:Heparin; D012685:Sepharose; C007789:fucoidan