A novel strategy to generate biologically active neo-glycosaminoglycan conjugates.
Heparin and heparan sulfate are structurally related polysaccharides with a variety of biological effects/functions. Most of these effects are due to interactions, of varying specificity, between the negatively charged polysaccharide chains and proteins. While such interactions generally involve a single saccharide domain of decasaccharide size or less, ternary complexes of two protein molecules binding to separate domains on a single polysaccharide chain are known to occur. To facilitate studies on domain organization and its importance for biological function a strategy was developed to chemically conjugate defined heparin oligomers in linear and chemoselective fashion. The procedure requires that the oligosaccharide to provide the reducing-terminal domain of the conjugate is generated by lyase degradation of a parent polysaccharide, whereas the nonreducing-terminal domain is obtained through deaminative cleavage with nitrous acid. The applicability of the method was demonstrated by constructing a conjugate composed of two heparin 12-mers, of which the reducing-terminal component contained the antithrombin-binding region, whereas the nonreducing-terminal domain did not. Contrary to any of the unconjugated oligomers, the product was found to efficiently promote the inactivation of thrombin by antithrombin.
Rong
J
J
Department of Medical Biochemistry and Microbiology, Box 582, The Biomedical Center, Uppsala University, S-751 23 Uppsala, Sweden.
Nordling
K
K
Björk
I
I
Lindahl
U
U
D000925:Anticoagulants / Q000494:pharmacology; D000991:Antithrombins / Q000737:chemistry / Q000494:pharmacology; D002850:Chromatography, Gel; D006025:Glycosaminoglycans / Q000737:chemistry* / Q000494:pharmacology; D006493:Heparin / Q000737:chemistry* / Q000494:pharmacology; D015394:Molecular Structure; D009844:Oligosaccharides / Q000737:chemistry; D009994:Osmolar Concentration; D013917:Thrombin / Q000037:antagonists & inhibitors
D000925:Anticoagulants; D000991:Antithrombins; D006025:Glycosaminoglycans; D009844:Oligosaccharides; D006493:Heparin; D013917:Thrombin