Mass spectral evidence for N-glycans with branching on fucose in a molluscan hemocyanin

Biochem Biophys Res Commun. 2005 Jun 3;331(2):562-70. doi: 10.1016/j.bbrc.2005.03.217.

Abstract

Glycopeptides, isolated from a trypsinolysate of functional unit (FU) RtH2-e of Rapana thomasiana hemocyanin subunit 2, were analysed by electrospray ionization mass spectrometry and MS/MS. From the molecular mass observed after deglycosylation, it was inferred that all glycopeptides shared the same peptide stretch 92-143 of FU RtH2-e with a glycosylation site at Asn-127. Besides the core structure Man(3)GlcNAc(2) for N-glycosylation, structures with a supplementary GlcNAc linked to either the Man(alpha1-3) or the Man(alpha1-6) arm and/or an additional tetrasaccharide unit connected to the other Man arm were observed, indicating the existence of microheterogeneity at the glycan level. The tetrasaccharide unit contains a central fucose moiety substituted with 3-O-methylgalactose and N-acetylgalactosamine, and linked to GlcNAc at the reducing end. This structure represents a novel N-glycan motif and is likely to be immunogenic. A second potential site for N-glycosylation in FU RtH2-e at Asn-17 was shown to be not glycosylated.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Carbohydrate Sequence
  • Fucose / analysis
  • Fucose / chemistry*
  • Glycopeptides / chemistry
  • Glycopeptides / isolation & purification
  • Glycopeptides / metabolism
  • Glycosylation
  • Hemocyanins / chemistry*
  • Mass Spectrometry
  • Molecular Sequence Data
  • Polysaccharides / analysis*
  • Polysaccharides / chemistry*
  • Snails / chemistry*
  • Trypsin / metabolism

Substances

  • Glycopeptides
  • Polysaccharides
  • Fucose
  • Hemocyanins
  • Trypsin