HPLC-ESI-MS and MS/MS structural characterization of multifucosylated N-glycoforms of the basic proline-rich protein IB-8a CON1+ in human saliva

Tiziana Cabras, Roberto Boi, Elisabetta Pisano, Federica Iavarone, Chiara Fanali, Sonia Nemolato, Gavino Faa, Massimo Castagnola, Irene Messana

Research output: Contribution to journalArticle

11 Citations (Scopus)

Abstract

This study describes the characterization of the glycan moieties and the peptide backbone of six glycoforms of IB-8a CON1(+), a basic proline-rich protein present in human saliva. MS analyses on the intact glycoproteins before and after N-deglycosylation with PNGase F and high-resolution MS/MS sequencing by LTQ Orbitrap XL of peptides and glycopeptides from tryptic digests allowed the structural characterization of the glycan moieties and the polypeptide backbone, as well as to establish the glycosylation site at the asparagine residue at 98th position. Five of the glycoforms carry a biantennary N-linked glycan fucosylated in the innermost N-acetylglucosamine of the core and showing from zero to four additional fucoses in the antennal region. The sixth glycoform carries a monoantennary monofucosylated oligosaccharide. The glycoform cluster was detected on 28 of 71 adult saliva specimens. Level of fucosylation showed interindividual variability with the major relative abundance for the trifucosylated glycoform. Nonglycosylated IB-8a CON1(+) and the variant IB-8a CON1(-), lacking of the glycosylation site, have been also detected in human saliva.
Original languageEnglish
Pages (from-to)1079-1086
Number of pages8
JournalJournal of Separation Science
Volume35
DOIs
Publication statusPublished - 2012

Keywords

  • Adolescent
  • Adult
  • Amino Acid Motifs
  • Chromatography, High Pressure Liquid
  • Female
  • Glycosylation
  • Humans
  • Male
  • Proteins
  • Saliva
  • Spectrometry, Mass, Electrospray Ionization
  • Tandem Mass Spectrometry
  • Young Adult

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