Neurofascin 186 is O-mannosylated within and outside of the mucin domain

J Proteome Res. 2012 Aug 3;11(8):3955-64. doi: 10.1021/pr200996y. Epub 2012 Jul 10.


Protein O-mannosylation is an important modification in mammals, and deficiencies thereof lead to a variety of severe phenotypes. Although it has already been shown that the amount of O-mannosyl glycans in brain is very high, only very few proteins have been identified as O-mannosylated. Additionally, the functions of the O-mannose-based glycans are still speculative and only investigated for α-dystroglycan. In a previous study a cis-located peptide was identified, which controls O-mannosylation in mammals. A BLAST search on the basis of this peptidic determinant identified other potential O-mannosylated proteins. Among these neurofascin was chosen for further analysis as a recombinant probe (mucin domain) and as an endogenous protein from mouse brain. Mass spectrometric data for both proteins confirmed that neurofascin186 is indeed O-mannosylated. Glycopeptide analysis by liquid chromatography-tandem mass spectrometry allowed for the identification of some of the O-mannosylation sites, which are not restricted to the mucin domain but were found also within N-terminal IgG and Fibronectin domains of the protein.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Motifs
  • Amino Acid Sequence
  • Animals
  • Carbohydrate Conformation
  • Carbohydrate Sequence
  • Cell Adhesion Molecules / chemistry
  • Cell Adhesion Molecules / isolation & purification
  • Cell Adhesion Molecules / metabolism*
  • Glycosylation
  • HEK293 Cells
  • Humans
  • Mannans / chemistry
  • Mannans / isolation & purification
  • Mannans / metabolism*
  • Mice
  • Mice, Inbred C57BL
  • Molecular Sequence Data
  • Mucins / chemistry
  • Nerve Growth Factors / chemistry
  • Nerve Growth Factors / isolation & purification
  • Nerve Growth Factors / metabolism*
  • Peptide Fragments / chemistry
  • Peptide Mapping
  • Protein Processing, Post-Translational*
  • Protein Structure, Tertiary
  • Recombinant Proteins / chemistry
  • Recombinant Proteins / isolation & purification
  • Recombinant Proteins / metabolism
  • Spectrometry, Mass, Electrospray Ionization


  • Cell Adhesion Molecules
  • Mannans
  • Mucins
  • Nerve Growth Factors
  • Nfasc protein, mouse
  • Peptide Fragments
  • Recombinant Proteins