Isolation of heparin-derived oligosaccharides containing 2-O-sulfated hexuronic acids, by lipoprotein lipase affinity chromatography

J Biochem Biophys Methods. 1996 Apr;32(1):27-32. doi: 10.1016/0165-022x(95)00045-s.

Abstract

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.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Borohydrides / metabolism
  • Chromatography, Affinity / methods*
  • Chromatography, Agarose
  • Chromatography, Thin Layer
  • Disaccharides / chemistry
  • Disaccharides / isolation & purification
  • Heparin / chemistry*
  • Hexuronic Acids / analysis*
  • Hydrolysis
  • Lipoprotein Lipase / metabolism*
  • Nitrous Acid
  • Oligosaccharides / chemistry
  • Oligosaccharides / isolation & purification*
  • Sulfates

Substances

  • Borohydrides
  • Disaccharides
  • Hexuronic Acids
  • Oligosaccharides
  • Sulfates
  • sodium borohydride
  • Heparin
  • Lipoprotein Lipase
  • Nitrous Acid