Domain structure of endothelial heparan sulphate.
The domain structure of heparan sulphate chains from an endothelial low-density proteoglycan was examined using specific degradations of the chains while attached to the intact proteoglycan. 'Inner' chain fragments, remaining on the protein core, were separated from 'outer' fragments by gel chromatography, and were subsequently released from the protein core by alkaline cleavage. The structure of 'inner' and 'outer' chain fragments was then examined and compared. Using deaminative cleavage we obtained evidence that the first N-sulphated glucosamine residue is variably positioned some 10-17 disaccharides from the xylose-serine linkage of the proteoglycan. Digestion with heparinase yielded 'inner' and 'outer' fragments covering a broad range of different sizes, indicating a scarce and variable distribution of sulphated iduronic acid in the native chains. N-sulphated glucosamine occurred more frequently in the 'outer' fragments. We also studied the affinity of the endothelial heparan sulphate chains towards two presumptive biological ligands, namely antithrombin III and lipoprotein lipase. A major part of the endothelial heparan sulphate chains showed a weak affinity for antithrombin III and the affinity was essentially lost on heparinase digestion. On lipoprotein lipase-agarose the endothelial heparan sulphate chains were eluted at the same salt concentration as heparin, and the binding persisted, although with decreased strength, after digestion with heparinase.
Lindblom
A
A
Department of Physiological Chemistry, University of Lund, Sweden.
Bengtsson-Olivecrona
G
G
Fransson
L A
LA
D002846:Chromatography, Affinity; D002850:Chromatography, Gel; D004730:Endothelium, Vascular / Q000737:chemistry*; D019763:Heparin Lyase; D006497:Heparan Sulfate / Q000737:chemistry*; D006801:Humans; D006868:Hydrolysis; D011133:Polysaccharide-Lyases / Q000737:chemistry; D011134:Polysaccharides / Q000737:chemistry
D019763:Heparin Lyase; D006497:Heparan Sulfate; D011133:Polysaccharide-Lyases; D011134:Polysaccharides