Research Article

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

  1. Taufika Islam Williams1,
  2. Diana A. Saggese1,
  3. Kristina L. Toups1,
  4. Jennifer L. Frahm1,
  5. Hyun Joo An2,
  6. Bensheng Li2,
  7. Carlito B. Lebrilla2,
  8. David C. Muddiman1,*

Article first published online: 6 MAR 2008

DOI: 10.1002/jms.1398

Issue

Journal of Mass Spectrometry

Journal of Mass Spectrometry

Volume 43, Issue 9, pages 1215–1223, September 2008

Additional Information

How to Cite

Williams, T. I., Saggese, D. A., Toups, K. L., Frahm, J. L., An, H. J., Li, B., Lebrilla, C. B. and Muddiman, D. C. (2008), Investigations with O-linked protein glycosylations by matrix-assisted laser desorption/ionization Fourier transform ion cyclotron resonance mass spectrometry. J. Mass Spectrom., 43: 1215–1223. doi: 10.1002/jms.1398

Author Information

  1. 1

    W.M. Keck FT-ICR Mass Spectrometry Laboratory, Department of Chemistry, North Carolina State University, Raleigh, NC 27695, USA

  2. 2

    University of California-Davis, Davis, CA 95616, USA

*W.M. Keck FT-ICR Mass Spectrometry Laboratory, Department of Chemistry, North Carolina State University, Raleigh, NC 27695-8204, USA.

Publication History

  1. Issue published online: 27 AUG 2008
  2. Article first published online: 6 MAR 2008
  3. Manuscript Accepted: 28 JAN 2008
  4. Manuscript Received: 25 OCT 2007

Funded by

  • North Carolina Biotechnology Center
  • LEAP Technologies, Inc.,
  • National Cancer Institute
  • National Institutes of Health. Grant Number: (R33 CA105295)
  • The W.M. Keck Foundation
  • North Carolina State University

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Keywords:

  • Fourier transform ion cyclotron mass spectrometry (FT-ICR-MS);
  • matrix-assisted laser desorption/ionization (MALDI);
  • mucin;
  • O-linked glycans;
  • protein glycosylations

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. Copyright © 2008 John Wiley & Sons, Ltd.

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