Skip to main content
SpringerLink
Log in

Matrix-assisted laser desorption/ionization tandem mass spectrometry and post-source decay fragmentation study of phenylhydrazones of N-linked oligosaccharides from ovalbumin

  • Articles
  • Published:
Journal of the American Society for Mass Spectrometry

Abstract

N-linked oligosaccharides were released from hen ovalbumin by PNGase F and derivatized with phenylhydrazine. They were then examined by matrix-assisted laser desorption/ionization (MALDI) mass spectrometry. Phenylhydrazones of N-glycans under MALDI-tandem mass spectrometry (MS/MS) and post-source decay (PSD) conditions produced relatively similar fragmentation patterns; however, more cross-ring cleavages and fragment ions corresponding to low abundance isomeric structures were detected by MS/MS and not in PSD. Most fragment ions corresponded to glycosidic cleavages with preferential loss of residues from the chitobiose core and the 3-antenna. Sialylated phenylhydrazone-N-glycans, characterized here for the first time in ovalbumin by tandem mass spectrometry, underwent losses of sialic acid residues followed the same fragmentation pathways observed with neutral derivatized glycans. The relative abundances of some fragment ions indicated the linkage position of sialic acid and provided information on the number of residues attached to the 6-antenna. Also, new structures of ovalbumin glycans were observed as part of this study and are reported here.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References

  1. Harvey, D. J.; Bateman, R. H.; Green, M. R. High-Energy Collision-Induced Fragmentation of Complex Oligosaccharides Ionized by Matrix-Assisted Laser Desorption/Ionization Mass Spectrometry. J. Mass Spectrom. 1997, 32, 167–187.

    Article  CAS  Google Scholar 

  2. Rasmussen, H. H.; Mortz, E.; Mann, M.; Roepstorff, P.; Celis, J. E. Identification of Transformation Sensitive Proteins Recorded in Human Two-Dimensional Gel Protein Databases by Mass Spectrometric Peptide Mapping Alone and in Combination with Microsequencing. Electrophoresis 1994, 15, 406–416.

    Article  CAS  Google Scholar 

  3. Reinhold, V. N.; Reinhold, B. B.; Costello, C. E. Carbohydrate Molecular Weight Profiling, Sequence, Linkage, and Branching Data: ES-MS and CID. Anal. Chem. 1995, 67, 1772–1784.

    Article  CAS  Google Scholar 

  4. Yamagaki, T.; Nakanishi, H. Ion Intensity Analysis of Post-Source Decay Fragmentation in Curved-Field Reflectron Matrix-Assisted Laser Desorption/Ionization Time-of-Flight Mass Spectrometry of Carbohydrates: For Structural Characterization of Glycosylation in Proteome Analysis. Proteomics 2001, 1, 329–339.

    Article  CAS  Google Scholar 

  5. Rouse, J. C.; Strang, A.-M.; Yu, W.; Vath, J. E. Isomeric Differentiation of Asparagine-Linked Oligosaccharides by Matrix-Assisted Laser Desorption/Ionization Post-Source Decay Time-of-Flight Mass Spectrometry. Anal. Biochem. 1998, 256, 33–46.

    Article  CAS  Google Scholar 

  6. Spengler, B.; Kirsch, D.; Kaufmann, R.; Lemoine, J. Structure Analysis of Branched Oligosaccharides Using Post-Source Decay in Matrix-Assisted Laser Desorption/Ionization Mass Spectrometry. J. Mass Spectrom. 1995, 30, 782–787.

    Article  Google Scholar 

  7. Yamagaki, T.; Ishizuka, Y.; Kawabata, S.; Nakanishi, H. Post-Source Decay Fragment Spectra of Cyclomalto-Octaose and Branched Cyclomalto-Hexaose by MALDI-TOF Mass Spectrometry. Rapid Commun. Mass Spectrom. 1996, 10, 1887–1890.

    Article  CAS  Google Scholar 

  8. Harvey, D. J. Matrix-Assisted Laser Desorption/Ionization Mass Spectrometry of Carbohydrates. Mass Spectrom. Rev. 1999, 18, 349–451.

    Article  CAS  Google Scholar 

  9. Harvey, D. J.; Naven, T. J. P.; Kuster, B.; Bateman, R. H.; Green, M. R.; Gritchley, G. Comparison of Fragmentation Modes for the Structural Determination of Complex Oligosaccharides Ionized by Matrix-Assisted Laser Desorption/Ionization Mass Spectrometry. Rapid Commun. Mass Spectrom. 1995, 9, 1556–1561.

    Article  CAS  Google Scholar 

  10. Harvey, D. J. Post-Source Decay Fragmentation of N-linked Carbohydrates from Ovalbumin and Related Glycoproteins. J. Am. Soc. Mass Spectrom. 2000, 11, 572–577.

    Article  CAS  Google Scholar 

  11. Domon, B.; Muller, D. R.; Richter, W. J. High Performance Tandem Mass Spectrometry for Sequence, Branching, and Interglycosidic Linkage Analysis of Peracetylated Oligosaccharides. Biomed. Environ. Mass Spectrom. 1990, 19, 390–392.

    Article  CAS  Google Scholar 

  12. Laine, R. A.; Yoon, E.; Mathier, T. J.; Abbas, S.; deLappe, B.; Jain, R.; Matta, K. Non-Reducing Terminal Linkage Position Determination in Intact and Permethylated Synthetic Oligosaccharides Having a Penultimate Amino Sugars: Fast Atom Bombardment Ionization, Collision-Induced Dissociation, and Tandem Mass Spectrometry. Biol. Mass Spectrom. 1991, 20, 505–514.

    Article  CAS  Google Scholar 

  13. Garrozzo, D.; Impallomeni, G.; Montaudo, G.; Spina, E. Structure of Underivatized Branched Oligosaccharides by Negative-Ion Fast-Atom Bombardment Mass Spectrometry. Rapid Commun. Mass Spectrom. 1992, 6, 550–552.

    Article  Google Scholar 

  14. Spengler, B.; Dolce, J. W.; Cotter, R. J. Infrared Laser Desorption Mass Spectrometry of Oligosaccharides: Fragmentation Mechanism and Isomer Analysis. Anal. Chem. 1990, 62, 1731–1737.

    Article  CAS  Google Scholar 

  15. Cornish, T. J.; Cotter, R. J. A Curved Field Reflectron Time-of-Flight Mass Spectrometry for the Simultaneous Focusing of Metastable Product Ions. Rapid Commun. Mass Spectrom. 1994, 8, 781–785.

    Article  CAS  Google Scholar 

  16. Hase, S.; Ibuki, T.; Ikenaka, T. Re-examination of the Pyridylamination Used for Fluorescence Labeling of Oligosaccharides and Its Application to Glycoproteins. J. Biochem. 1984, 95, 197–203.

    CAS  Google Scholar 

  17. Poulter, L.; Burlingame, A. L. Desorption Mass Spectrometry of Oligosaccharides Coupled with Hydrophobic Chromophores. Methods Enzymol 1990, 193, 661–689.

    Article  CAS  Google Scholar 

  18. Okamoto, M.; Takahashi, K. I.; Doi, T. Sensitive Detection and Structural Characterization of Trimethyl(p-aminophenyl)-Ammonium-Derivatized Oligosaccharides by Electrospray Ionization-Mass Spectrometry and Tandem Mass Spectrometry. Rapid Commun. Mass Spectrom. 1995, 9, 641–643.

    Article  CAS  Google Scholar 

  19. Hase, S. Precolumn Derivatization for Chromatographic and Electrophoretic Analysis of Carbohydrates. J. Chromatogr. A 1996, 720, 173–182.

    Article  CAS  Google Scholar 

  20. Naven, T. J. P.; Harvey, D. J. Cationic Derivatization of Oligosaccharides with Girard’s T Reagent for Improved Performance in Matrix-Assisted Laser Desorption/Ionization and Electrospray Mass Spectrometry. Rapid Commun. Mass Spectrom. 1996, 10, 829–834.

    Article  CAS  Google Scholar 

  21. Strydom, D. J. Chromatographic Separation of 1-Phenyl-3-Methyl-5-Pyrazolone-Derivatized Neutral, Acidic, and Basic Aldoses. J. Chromatogr. A 1994, 678, 17.

    Article  CAS  Google Scholar 

  22. Shen, X.; Perreault, H. Characterization of Carbohydrates Using a Combination of Derivatization, High-Performance Liquid Chromatography and Mass Spectrometry. J. Chromatogr. A 1998, 811, 47–59.

    Article  CAS  Google Scholar 

  23. Lattova, E.; Perreault, H. Labeling Saccharides with Phenylhydrazine for Electrospray and Matrix-Assisted Laser Desorption/Ionization Mass Spectrometry. J. Chromatogr. B 2003, 793, 167–179.

    Article  CAS  Google Scholar 

  24. Yoshino, K. I.; Takato, T.; Muruta, H.; Shimonishi, Y. Use of the Derivatizing Agent 4-Aminobenzoic Acid 2-(Diethylamino)Ethyl Ester for High-Sesitivity Detection of Oligosaccharides by Electrospray Ionization Mass Spectrometry. Anal. Chem. 1995, 67, 4028–4031.

    Article  CAS  Google Scholar 

  25. Okafo, G.; Burrow, L.; Carr, S. A.; Roberts, G. D.; Johnson, W.; Camilleri, P. A Coordinated High-Performance Liquid Chromatographic, Capillary Electrophoretic, and Mass Spectrometric Approach for the Analysis of Oligosaccharide Mixtures with 2-Aminoacridone. Anal. Chem. 1996, 68, 4424–4430.

    Article  CAS  Google Scholar 

  26. Shen, X.; Perreault, H. Electrospray Ionization Mass Spectrometry of 1-Phenyl-3-Methyl-5-Pyrazolone Derivatives of Neutral and N-Acetylated Oligosaccharides. J. Mass Spectrom. 1999, 34, 502–510.

    Article  CAS  Google Scholar 

  27. Saba, J. A.; Shen, X.; Jamieson, J. C.; Perreault, H. Investigation of Different Combinations of Derivatization, Separation Methods, and Electrospray Ionization Mass Spectrometry for Standard Oligosaccharides and Glycans from Ovalbumin. J. Mass Spectrom. 2001, 36, 563–574.

    Article  CAS  Google Scholar 

  28. Lattova, E.; Perreault, H. Profiling of N-Linked Oligosaccharides Using Phenylhydrazine Derivatization and Mass Spectrometry. J. Chromatog. A. 2003, 1016, 71–87.

    Article  CAS  Google Scholar 

  29. Tomyia, N.; Awaya, J.; Kurono, M.; Endo, S.; Arata, Y. Takahashi, N. Analysis of N-Linked Oligosaccharides Using a Two-Dimensional Mapping Technique. Anal. Biochem. 1988, 171, 73–90.

    Article  Google Scholar 

  30. Da Silva, M. L. C.; Stubbs, H. J.; Tamura, T.; Rice, K. G. 1H-NMR Characterization of a Hen Ovalbumin Tyrosinamide N-Linked Oligosaccharide Library. Arch. Biochem. Biophys. 1995, 318, 465–475.

    Article  Google Scholar 

  31. Tarentino, A. L.; Plummer, T. H. Enzymatic Deglycosylation of Asparagine-Linked Glycans: Properties and Specificity of Oligosaccharide Cleaving Enzymes from Flavobacterium meningosepticum. Methods in Enzymology 1994, 230, 44–57.

    Article  CAS  Google Scholar 

  32. Tarentino, A. L.; Gomez, C. M.; Plummer, T. H. Deglycosylation of Asparagines-Linked Glycans by Peptide N-Glycosidase F. Biochemistry 1985, 24, 4665–4671.

    Article  CAS  Google Scholar 

  33. Loboda, A. V.; Krutchinsky, N. N.; Bromirski, M. P.; Ens, W.; Standing, K. G. A Tandem Quadrupole/Time-of-Flight Mass Spectrometer with a Matrix-Assisted Laser Desorption/Ionization Source: Design and Performance. Rapid Commun. Mass Spectrom. 2000, 14, 1047–1057.

    Article  CAS  Google Scholar 

  34. Domon, B.; Costello, C. E. A Systematic Nomenclature for Carbohydrate Fragmentations in FABMS/MS Spectra of Glycoconjugates. Glycoconj. J. 1988, 5, 397–409.

    Article  CAS  Google Scholar 

  35. Yamashita, K.; Kamerling, J. P.; Kobata, A. Structural Studies of the Sugar Chains of the Carbohydrate Moiety of Hen Ovomucoid: Occurrence of a Series of Pentaantennary Complex-Type Asparagines-Linked Sugar Chains. J. Biol. Chem. 1982, 257, 12809–12814.

    CAS  Google Scholar 

  36. Domingues, R. M. M.; Marques, M. G. O. S.; Vale, C. A. V.; Neves, M. G.; Cavaleiro, J. A. S.; Ferrer-Correia, A. J.; Nemirovskiy, O. V.; Gross, M. L. Do Charge-Remote Fragmentations Occur Under Matrix-Assisted Laser Desorption Ionozation Post-Source Decomposition and Matrix-Assisted Laser Desorption Ionization Collisionally Activated Decomposition?. J. Am. Soc. Mass Spectrom. 1999, 10, 217–223.

    Article  Google Scholar 

  37. Harvey, D. J.; Wing, D. R.; Kuster, B.; Wilson, I. B. H. Composition of N-Linked Carbohydrates from Ovalbumin and Copurified Glycoproteins. J. Am. Soc. Mass Spectrom. 2000, 11, 564–571.

    Article  CAS  Google Scholar 

  38. Yamashita, K.; Tachibana, Y.; Hitoi, A.; Kobata, A. Sialic Acid-Containing Sugar Chains of Hen Ovalbumin and Ovomucoid. Carbohydr. Res. 1984, 130, 271–288.

    Article  CAS  Google Scholar 

Download references

Author information

Author notes

  1. Erika Lattova

    Present address: Institute of Chemistry, Slovak Academy of Sciences, 842 38, Bratislava, Slovakia

Authors and Affiliations

  1. Chemistry Department, University of Manitoba, 144 Dysart Road, R3T 2N2, Winnipeg, Manitoba, Canada

    Erika Lattova & Hélène Perreault

  2. Physics and Astronomy Department, University of Manitoba, Winnipeg, Manitoba, Canada

    Oleg Krokhin

Authors
  1. Erika Lattova
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Hélène Perreault
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Oleg Krokhin
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Hélène Perreault.

Additional information

Published online March 12, 2004

Rights and permissions

Reprints and permissions

About this article

Cite this article

Lattova, E., Perreault, H. & Krokhin, O. Matrix-assisted laser desorption/ionization tandem mass spectrometry and post-source decay fragmentation study of phenylhydrazones of N-linked oligosaccharides from ovalbumin. J Am Soc Mass Spectrom 15, 725–735 (2004). https://doi.org/10.1016/j.jasms.2004.01.012

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1016/j.jasms.2004.01.012

Keywords

Search

Navigation