title: Preparation of heparin/heparan sulfate oligosaccharides with internal N-unsubstituted glucosamine residues for functional studies.
abstract: The rare N-unsubstituted glucosamine (GlcNH (3)(+)) residues in heparan sulfate (HS) have important biological and pathophysiological roles. However, it is difficult to prepare naturally-occuring, GlcNH(3)(+)-containing oligosaccharides from HS because of their low abundance, as well as the inherent problems in both excising and identifying them. Therefore, the ability to chemically generate a series of structurally-defined oligosaccharides containing GlcNH(3)(+) residues would greatly contribute to investigating their natural role in HS. In this study, a series of heparin/HS oligosaccharides, from dp6 up to dp16 in length that possess internal GlcNH(3)(+) residues were prepared by a combination of chemical modification and heparinase I digestion. Purification and structural analysis of the major species derived from the octa- to dodeca-saccharide size fractions indicated the introduction of between 1 and 3 internal GlcNH(3)(+) residues per oligosaccharide. In addition, a GlcNH(3)(+) residue was selectively introduced into an internal position in a tetrasaccharide species by direct chemical modification. This selectivity has potential as an alternative procedure for preparing internally-modified oligosaccharides of various lengths. The utility of such oligosaccharides was demonstrated by a comparison of the binding of three different tetrasaccharide species containing 0, 1 and 2 free amino groups to the NK1 truncated variant of hepatocyte growth factor/scatter factor.
authors.affiliation: Institute of Glycobiochemistry, National Engineering Research Centre of Chemical Fertilizer Catalyst, Fu Zhou University, Fu Zhou 350002, People's Republic of China. zheng0wei@hotmail.com
authors.forename: Zheng
authors.initials: Z
authors.lastname: Wei
authors.forename: Jon Andrew
authors.initials: JA
authors.lastname: Deakin
authors.forename: Bärbel Sabine
authors.initials: BS
authors.lastname: Blaum
authors.forename: Dušan
authors.initials: D
authors.lastname: Uhrín
authors.forename: John Thomas
authors.initials: JT
authors.lastname: Gallagher
authors.forename: Malcolm
authors.initials: M
authors.lastname: Lyon
mesh_terms: D000818:Animals; D002417:Cattle; D005591:Chemical Fractionation; D002850:Chromatography, Gel; D002851:Chromatography, High Pressure Liquid; D004187:Disaccharides / Q000032:analysis; D005944:Glucosamine / Q000378:metabolism*; D006493:Heparin / Q000031:analogs & derivatives / Q000138:chemical synthesis* / Q000737:chemistry; D006497:Heparan Sulfate / Q000138:chemical synthesis* / Q000737:chemistry; D017228:Hepatocyte Growth Factor / Q000378:metabolism; D006801:Humans; D009682:Magnetic Resonance Spectroscopy; D009844:Oligosaccharides / Q000138:chemical synthesis* / Q000737:chemistry / Q000302:isolation & purification; D011485:Protein Binding; D015238:Sulfotransferases / Q000378:metabolism
chemical_list: D004187:Disaccharides; D005944:Glucosamine; D006493:Heparin; D006497:Heparan Sulfate; D017228:Hepatocyte Growth Factor; D009844:Oligosaccharides; D015238:Sulfotransferases; C044153:heparin, N-desulfated