Structure of heparin-derived tetrasaccharides

Abstract

The structure of heparin was examined by characterizing a disaccharide and five of the more than a dozen tetrasaccharide components obtained by its depolymerization with flavobacterial heparinase. Enzymic depolymerization of porcine mucosal heparin results in a mixture of di-, tetra-, hexa- and higher oligo-saccharides. The di- and tetra-saccharide components represent 75mol/100mol of these heparin fragments. Ion-exchange chromatography indicates the presence of only one disaccharide, deltaIdu2S(1----4)-alpha-D-GlcNS6S (where Idu is iduronic acid, deltaIdu is 4-deoxy-alpha-L-threo-hex-4-enopyranosyluronic acid, GlcN is glucosamine, GlcA is glucuronic acid and S is sulphate), but results in the isolation of five major and at least seven minor tetrasaccharide components. The structures of the disaccharide and five major tetrasaccharides were determined by chemical, enzymic, electrophoretic and spectroscopic methods, including 13C, 1H n.m.r. and fast atom bombardment-m.s. The structure of these five tetrasaccharides are: delta Idu2S(1----4)-alpha-D-GlcNS6S(1---4)-alpha-L-Idu2S(1-- --4)-alpha -D-GlcNS6S; delta Idu2S(1----4)-alpha-D-GlcNS6S(1----4)-beta-D-GlcA(1--- -4)- alpha-D-GlcNS6S; delta Idu2S(1----4)-alpha-D-GlcNS(1----4)-beta-D-GlcA delta Idu2S(1----4)-alpha-D-GlcNAc(1----4)-beta-D-GlcA(1----4)- alpha-D-GlcNS6S; and delta Idu2S(1----4)-alpha- D-GlcNAc(1----4)-alpha-L-Idu(1----4)-alpha-D-GlcNS6S. Biological activity for the disaccharide and the five major tetrasaccharides was examined, and none of them were found to possess significant anticoagulant activity.

The Responses of Hyperglycemic Dividing Mesangial Cells to Heparin Are Mediated by the Non-reducing Terminal Trisaccharide

“…To determine whether exposing the non-reducing GlcNS (6S) is required to initiate the hyaluronan and monocyte-adhe- sive responses, a fully sulfated ⌬-tetrasaccharide (⌬-Tetra) purified from heparin lyase-digested heparin was tested (23). FACE analysis of the ⌬-Tetra showed a single band that was converted to fully sulfated ⌬-UA(2S)-GlcNS(6S) disaccharides after heparin lyase digestion (Fig.…”

On‐line size‐exclusion chromatography/mass spectrometry of low molecular mass heparin

“…3) and a fraction of heparin tetrasaccharides mainly composed by hexasulphated heparin tetrasaccharide and pentasulphated heparin tetrasaccharide. The ESI ionization parameters needed to be set very carefully to prevent the loss of SO 3 , since this will result in determination of a too low number of sulphate groups.…”

“…The first successful MS analysis of small heparin oligosaccharides was achieved using fast atom bombardment (FAB) ionization, where the oligosaccharides were ionized as salt adducts, 3,4 and 252 Cf plasma desorption (PD) ionization, where the oligosaccharides were complexed with tridodecylmethylammonium prior to the MS analysis. 5 In both the case of FAB and PD, a considerable SO 3 loss took place during the MS analysis, making determination of the degree of sulphation of heparin oligosaccharides in mixtures difficult.…”

A Major Common Trisulfated Hexasaccharide Core Sequence, Hexuronic Acid(2-Sulfate)-Glucosamine(N-Sulfate)-Iduronic Acid-N-Acetylglucosamine-Glucuronic Acid-Glucosamine(N-Sulfate), Isolated from the Low Sulfated Irregular Region of Porcine Intestinal Heparin

“…The internal uronic acid residues of each isolated oligosaccharide were unambiguously identified based upon the chemical shifts of the anomeric proton signals and the coupling constants J 1,2 . Anomeric proton signals of an ␣IdceA and a ␤GlcA residue in heparin/ heparan sulfate oligosaccharides are observed around ␦ 5.2-5.0 and 4.7-4.5, respectively (22,35). The coupling constants J 1,2 of ␣IdceA and ␤GlcA in heparin/heparan sulfate oligosaccharides are approximately 3.0 and 8.0 Hz, respectively (22,36).…”