Heparan sulphate/heparin glycosaminoglycans with strong affinity for the growth-promoter spermine have high antiproliferative activity

摘要: Depletion of intracellular polyamine pools inhibits cell proliferation. Polyamine are maintained by synthesis and uptake from the extracellular environment. It may be expected that cationic polyamines sequestered polyanionic glycosaminoglycan substituents proteoglycans. Moreover, highly sulphated heparin-related glycans inhibit growth human embryonic lung fibroblasts. We have therefore investigated interactions between glycosaminoglycans. Affinity chromatography various on heparin-agarose indicated spermine was only bound efficiently to this type glycan. By using equilibrium dialysis we found binds a heparan sulphate/heparin preparation with dissociation constant 3.7 x 10(-5)M. Enzymatic degradation sulphate three different lyases, separately or in combination absence presence spermine, used generate spermine-binding degradation-protected oligosaccharides. As chromatographic electrophoretic analysis size- chargeheterogeneous collection obtained. However, protected oligosaccharides derived antiproliferative sulphates were inactive. Highly sulphated, subfractionated spermine-agarose yielding high-affinity material increased activity. A very potent obtained pig skin. Although there generally clear correlation high spermine-affinity strong growth-inhibition, no content oligosaccharide mapping patterns could detected. Beef comprised naturally occurring fragments eicosasaccharide size substantially specific these longer than generated enzymatic (hexa- tetradecasaccharide), multiple sites tandem necessary induce