Identification and structural characterization of a mannose-6-phosphate containing oligomannosidic N-glycan from human erythropoietin secreted by recombinant BHK-21 cells

FEBS Lett. 1995 May 29;365(2-3):203-8. doi: 10.1016/0014-5793(95)00473-m.

Abstract

A sialidase resistant mono-charged N-glycan was isolated from glycosylation site I (Asn-24) of recombinant human erythropoietin expressed from baby hamster kidney (BHK-21) cells and constituted approximately 2-4% of the oligosaccharide material at this glycosylation site. Mass spectrometry and both 1- and 2-dimensional NMR techniques revealed a high mannose type structure (Man6) with a phospho-diesterbridged N-acetylglucosamine as follows: [formula: see text]

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Carbohydrate Conformation
  • Carbohydrate Sequence
  • Cell Line
  • Chromatography, High Pressure Liquid
  • Cricetinae
  • Erythropoietin / biosynthesis
  • Erythropoietin / chemistry*
  • Glycosylation
  • Humans
  • Kidney
  • Magnetic Resonance Spectroscopy
  • Mannosephosphates / analysis*
  • Mass Spectrometry
  • Methylation
  • Molecular Sequence Data
  • Oligosaccharides / chemistry*
  • Oligosaccharides / isolation & purification
  • Recombinant Proteins / biosynthesis
  • Recombinant Proteins / chemistry
  • Transfection

Substances

  • Mannosephosphates
  • Oligosaccharides
  • Recombinant Proteins
  • Erythropoietin
  • mannose-6-phosphate