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 and interactions. The possible presence of a unique binding site for ATIII and HCII, on each sulphated polysaccharide, was also studied.     

Changes of antithrombin III (AT III) in AT III deficient patients during long-term anticoagulant treatment

Antithrombin III deficient patients with manifest thromboembolic diseases need long term coumarin treatment. There are contradictory data on the change of AT III during this therapy. The authors observed 5 patients with severe AT III decrease type I, 3 with functional abnormality and 2 with a pathological heparin binding. AT III function was determined by the Gerendás-Rák method and with chromogenic substrate. AT III antigen was measured with Behring M-Partigen and Laurell rocket electrophoresis. Crossed immunoelectrophoresis was carried out in all patients. In patients with type I AT III decrease, AT III hasn't changed even in a long period of more than 10 years. In the other types AT III became normal. The pathological heparin binding wasn't changed.     

Effect of alpha 1-proteinase inhibitor and sulphated polysaccharides on the activity of m beta-acrosin

Purified m beta-acrosin catalysed amidolysis in vitro of several p-nitroanilides with C-terminal arginine residues. alpha 1-proteinase inhibitor inhibited amidolysis catalysed by the enzyme. This effect of alpha 1-Proteinase inhibitor was not prevented by pre-incubation of the enzyme with heparin or any other glycosaminoglycan. Pre-incubation of the enzyme with sulphated dextran or sulphated cellulose alleviated the effect of alpha 1-proteinase inhibitor. These results are discussed in terms of possible in vivo modulation by alpha 1-proteinase inhibitor of acrosin activity.     

The effect of growth factors on the cytotoxicity of sulphated polysaccharides

The cytotoxicity of sulphated polysaccharides on 3T3-L1 fibroblasts was investigated. Dextran sulphate with a molecular weight of more than 100000 and with a number of sulphate groups per sugar unit (degree of sulphation) of 2.4 showed strong cytotoxicity to 3T3-L1 fibroblasts. The synthetic (1→6)-- -mannopyranan sulphate having a degree of sulphation of more than 1.0 and with a molecular weight of approx. 100000 also exhibited cytotoxicity. The results indicated that the cytotoxicity of sulphated polysaccharides strongly depended on their molecular weight and degree of sulphation. FGF-1 (aFGF) and FGF-2 (bFGF) protected the cell from the damage caused by the sulphated polysaccharide, while PDGF, EGF, and HGF had no such effect. Therefore, it is suggested that the rescuing effect is one of the special biological activities, and is shown only by FGFs.     

Localization of sulphated polysaccharides by X-ray microanalysis in Laminaria saccharina

Summary Analysis of thick sections of Laminaria thallus by X-ray electron microscope microanalysis using EMMA 4 shows that sulphated polysaccharide is located in secretory canals, the middle lamella of cell walls and on the thallus surface. Sulphation occurs in the Golgi bodies of specialised secretory cells. These results confirm previous reports using histochemical and autoradiographic techniques.