Investigations with O-linked protein glycosylations by matrix-assisted laser desorption/ionization Fourier transform ion cyclotron resonance mass spectrometry

Taufika Islam Williams, Diana A. Saggese, Kristina L. Toups, Jennifer L. Frahm, Hyun Joo An, Bensheng Li, Carlito B Lebrilla, David C. Muddiman

Research output: Contribution to journalArticle

12 Scopus citations

Abstract

Posttranslational modifications such as glycosylation can play a fundamental role in signaling pathways that transform an ordinary cell into a malignant one. The development of a protocol to detect these changes in the preliminary stages of disease can lead to a sensitive and specific diagnostic for the early detection of malignancies such as ovarian cancer in which differential glycan patterns are linked to etiology and progression. Small variations in instrument parameters and sample preparation techniques are known to have significant influence on the outcome of an experiment. For an experiment to be effective and reproducible, these parameters must be optimized for the analyte(s) under study. We present a detailed examination of sample preparation and matrix-assisted laser desorption/ionization Fourier transform ion cyclotron resonance mass spectrometry (MALDI-FT-ICR-MS) analysis of O-linked glycans globally cleaved from mucin glycoproteins. Experiments with stable isotope-labeled biomolecules allowed for the establishment of appropriate acquisition times and excitation voltages for MALDI-FT-ICR-MS of oligosaccharides. Quadrupole ion guide optimization studies with mucin glycans identified conditions for the comprehensive analysis of the entire mass range of O-linked carbohydrates in this glycoprotein. Separately optimized experimental parameters were integrated in a method that allowed for the effective study of O-linked glycans.

Original languageEnglish (US)
Pages (from-to)1215-1223
Number of pages9
JournalJournal of Mass Spectrometry
Volume43
Issue number9
DOIs
StatePublished - Sep 2008

Keywords

  • Fourier transform ion cyclotron mass spectrometry (FT-ICR-MS)
  • Matrix-assisted laser desorption/ionization (MALDI)
  • Mucin
  • O-linked glycans
  • Protein glycosylations

ASJC Scopus subject areas

  • Spectroscopy

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