Qualitative and quantitative comparison of N-glycans between pig endothelial and islet cells by high-performance liquid chromatography and mass spectrometry-based strategy

J Mass Spectrom. 2009 Jul;44(7):1087-104. doi: 10.1002/jms.1587.

Abstract

N-glycan structures released from miniature pig endothelial and islet cells were determined by matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF), negative ion electrospray ionization (ESI) MS/MS and normal-phase high performance liquid chromatography (NP-HPLC) combined with exoglycosidase digestion. Totally, the identified structures were 181 N-glycans including 129 sialylated and 18 alpha-galactosylated glycans from pig endothelial cells and 80 N-glycans including 41 sialylated and one alpha-galactosylated glycans from pig islet cells. The quantity of the alpha-galactosylated glycans from pig islet cells was certainly neglectable compared to pig endothelial cells. A number of NeuGc-terminated N-glycans (80 from pig endothelial cells and 13 from pig islet cells) are newly detected by our mass spectrometric strategies. The detailed structural information will be a matter of great interest in organ or cell xenotransplantation using alpha 1,3-galactosyltransferase gene-knockout (GalT-KO) pig.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Cell Line
  • Chromatography, High Pressure Liquid / methods*
  • Endothelial Cells / chemistry*
  • Endothelial Cells / metabolism
  • Glycomics / methods*
  • Glycoside Hydrolases / metabolism
  • Islets of Langerhans / chemistry*
  • Islets of Langerhans / metabolism
  • Polysaccharides / chemistry*
  • Polysaccharides / metabolism
  • Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / methods*
  • Swine

Substances

  • Polysaccharides
  • Glycoside Hydrolases