Different fates of the oligosaccharide moieties of lipid intermediates

Glycobiology. 1992 Apr;2(2):127-36. doi: 10.1093/glycob/2.2.127.

Abstract

We have previously described that the N-glycosylation process was accompanied by the release of oligosaccharide-phosphates and neutral oligosaccharides. The relationship between oligosaccharide-P-P-dolichol and its metabolic products (glycoproteins, oligosaccharide-phosphates and neutral oligosaccharides) was investigated by analysing the structure of the oligosaccharide moieties and the kinetic behaviour of the various species in pulse and pulse/chase experiments. For these studies, a glycosylation mutant of Chinese hamster ovary cells (B3F7) which does not synthesize mannosylphosphoryldolichol was utilized. Evidence was obtained for the presence of two pools of oligosaccharide-P-P-dolichol which have different fates. One pool is not glucosylated, is rapidly labelled and immediately chased by mannose, and generates the oligosaccharide-phosphate species. The second pool is glucosylated, exhibits a lag time (5-10 min) prior to being labelled, and is utilized in the glycosylation of proteins and in the production of neutral oligosaccharides. We postulate that the cleavage of non-glycosylated lipid intermediates generating oligosaccharide-phosphates represents a 'bypass' in the dolichol cycle which allows direct regeneration of dolichyl phosphate. The other metabolic fate of non-glucosylated oligosaccharide-lipids, glucosylation, results in their use as effective substrates for the glycosylation of proteins or in the generation of neutral oligosaccharides.

Publication types

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

MeSH terms

  • Animals
  • CHO Cells / metabolism*
  • Chromatography, High Pressure Liquid
  • Cricetinae
  • Dolichol Monophosphate Mannose / chemistry
  • Dolichol Phosphates / chemistry
  • Glycosylation
  • Hydrogen-Ion Concentration
  • Lipid Metabolism*
  • Lipids / chemistry
  • Oligosaccharides / chemistry
  • Oligosaccharides / metabolism*
  • Rats

Substances

  • Dolichol Phosphates
  • Lipids
  • Oligosaccharides
  • Dolichol Monophosphate Mannose