Isolation of heparin-derived oligosaccharides containing 2-O-sulfated hexuronic acids, by lipoprotein lipase affinity chromatography.
Oligosaccharides (hexa to dodeca) terminating with [3H]2,5-anhydromannitol (AManR) were isolated from heparin by partial cleavage with nitrous acid at low pH (pH 1.5) followed by gel filtration and reduction with [3H]NaBH4. They were subsequently chromatographed on a lipoprotein lipase (LpL)-Sepharose column. High- and low-affinity oligosaccharides for LpL were isolated and characterized. Disaccharide analysis revealed the presence of (IdceA(2-SO4)-->AManR6-SO4) and (IdceA(2-SO4)-->AManR) as the major disaccharide products after low pH nitrous acid treatment. The oligosaccharides are, therefore, enriched in IdceA(2-SO4)-(GlcNSO4 +/- 6-SO4) sequences. Furthermore, they are found to be composed of 2-O-sulfated hexuronic acid-containing sequences, structural features, characteristic of heparin and heparan sulfate oligosaccharides with potential antiproliferative activities. These oligosaccharides may have the potential as lipase-releasing agents from endothelial and adipocyte surfaces.
Parthasarathy
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Department of Comparative Medicine, Bowman Gray School of Medicine, Wake Forest University, Winston-Salem, NC 27157-1040, USA.
Goldberg
I J
IJ
Sivaram
P
P
Wagner
W D
WD
D001894:Borohydrides / Q000378:metabolism; D002846:Chromatography, Affinity / Q000379:methods*; D002847:Chromatography, Agarose; D002855:Chromatography, Thin Layer; D004187:Disaccharides / Q000737:chemistry / Q000302:isolation & purification; D006493:Heparin / Q000737:chemistry*; D006603:Hexuronic Acids / Q000032:analysis*; D006868:Hydrolysis; D008071:Lipoprotein Lipase / Q000378:metabolism*; D009608:Nitrous Acid; D009844:Oligosaccharides / Q000737:chemistry / Q000302:isolation & purification*; D013431:Sulfates
D001894:Borohydrides; D004187:Disaccharides; D006603:Hexuronic Acids; D009844:Oligosaccharides; D013431:Sulfates; C025364:sodium borohydride; D006493:Heparin; D008071:Lipoprotein Lipase; D009608:Nitrous Acid