Mapping and quantification of the major oligosaccharide components of heparin.
A new method of determining the oligosaccharide composition of commercial glycosaminoglycan heparin is described in which heparin was first depolymerized using heparin lyase (EC 4.2.2.7), and then analysed by a single h.p.l.c. step. All 20 of the porcine and bovine heparins examined were found to contain a small number of major oligosaccharide components, which on average comprised 86% of their mass. The five most abundant oligosaccharides have defined chemical structures. Although the relative abundance of oligosaccharides varied, the heparins examined were surprisingly similar. Porcine, bovine, low-Mr, and high and low antithrombin III (ATIII)-affinity heparins, however, each had distinctly different proportions of these major oligosaccharide components. The concentrations of one of these five oligosaccharides, containing a portion of the ATIII binding site, correlated with the anticoagulant activity of the ATIII-affinity-fractionated porcine-mucosal heparins from which it was derived. An additional oligosaccharide of undetermined structure was found in significant quantities in both bovine heparin and high ATIII-affinity porcine-mucosal heparin. The correlation between oligosaccharide concentration and anticoagulant activity suggests that the oligosaccharide is derived from a structural variant of the ATIII-binding site. Finally, for the heparins examined chondroitin/dermatan sulphate formed 0.6-7.4% of their mass.
Linhardt
R J
RJ
Division of Medicinal and Natural Products Chemistry, College of Pharmacy, University of Iowa, Iowa City 52242.
Rice
K G
KG
Kim
Y S
YS
Lohse
D L
DL
Wang
H M
HM
Loganathan
D
D
D000818:Animals; D000990:Antithrombin III / Q000378:metabolism; D001665:Binding Sites; D002417:Cattle; D002851:Chromatography, High Pressure Liquid; D006493:Heparin / Q000378:metabolism; D008722:Methods; D009844:Oligosaccharides / Q000032:analysis*; D013552:Swine
D009844:Oligosaccharides; D000990:Antithrombin III; D006493:Heparin