Structural characterization of heparin's binding domain for human platelets.
The structural features of heparin that are involved in binding to human platelets were investigated by a competitive binding approach. A range of heparin-derived glycosaminoglycans (GAGs) with relatively defined structure were prepared by different methods of depolymerization of pharmaceutical heparin, followed by fractionation according to molecular weight and net charge. Competitive binding to platelets was dependent on molecular weight but not on the net charge of the GAGs. The method for depolymerization significantly affected the binding activity of the resulting GAG. Heparinase I and nitrous acid depolymerization produced GAGs with lower binding affinity for platelets than those GAGs derived from the treatment with periodate followed by alkali. The IC20 (concentration producing 20% inhibition of binding) was 0.05 microM for unfractionated heparin, 0.11 microM for a periodate treated GAG, and 2 microM for comparably sized GAGs (M(r) approximately 6,000-8,000) derived by heparinase I or nitrous acid treatment. Thus, the disaccharide units GlcNSO3-6S--IdoA-2S or GlcNSO3--IdoA-2S [(2-deoxy-2-sulfoamido-6-O-sulfo-alpha-D-glycopyranosyl)-(1- 4)-O-(2-O-sulfo-alpha-L-idopyranosyluronic acid) or (2-deoxy-2-sulfoamido-alpha-D-glycopyranosyl)-(1-4)-O-(2-O-s ulfo-alpha-L-idopyranosyluronic acid)] may be crucial elements for binding to the platelet, because these are known to be preserved during periodate/alkali treatment, but readily decomposed by heparinase I and nitrous acid. Understanding this structural specificity for platelet binding may be useful for the development of heparins with high or low platelet reactivity.
Department of Chemistry, Faculty of Science, Osaka University, Japan.
Y
Y
Suda
D
D
Marques
J C
JC
Kermode
S
S
Kusumoto
M
M
Sobel
D001665:Binding Sites; D001667:Binding, Competitive; D001792:Blood Platelets / Q000378:metabolism*; D002240:Carbohydrate Sequence; D006025:Glycosaminoglycans / Q000737:chemistry* / Q000378:metabolism; D006493:Heparin / Q000737:chemistry* / Q000378:metabolism; D019763:Heparin Lyase; D006801:Humans; D008958:Models, Molecular; D008969:Molecular Sequence Data; D011133:Polysaccharide-Lyases
D006025:Glycosaminoglycans; D006493:Heparin; D011133:Polysaccharide-Lyases; D019763:Heparin Lyase