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The high mannose glycans from bovine ribonuclease B isomer characterization by ion trap MS

  • Justin M. Prien
  • David J. Ashline
  • Anthony J. Lapadula
  • Hailong Zhang
  • Vernon N. Reinhold
Articles

Abstract

Thirteen high mannose isomers have been structurally characterized within three glycomers, Man5GlcNAc2, Man7GlcNAc2, and Man8GlcNAc2 released from bovine ribonuclease B, six previously unreported. The study was carried out with a single ion trap instrument involving no chromatography. Three previously characterized isomers from Man7 and Man8 (three each) have been identified plus one unreported Man7 isomer. Incomplete α-glucosidase activity on the Man6 and Man7 glycoproteins appears to account for two additional isomeric structures. The preeminence of ion traps for detail analysis was further demonstrated by resolving three new isomers within the Man5 glycomer summing to the six previously unreported structures in this glycoprotein. All reported structures represent a distribution of Golgi processing remnants that fall within the Man9GlcNAc2 footprint. Topologies were defined by ion compositions along a disassembly pathway while linkage and branching were aided by spectral identity in a small oligomer fragment library. Isomers from this glycoprotein appear to represent a distribution of Golgi processing remnants, and an alphanumeric classification scheme has been devised to identify all products. Although numerous analytical strategies have been introduced to identify selected components of structure, it has been the continued focus of this and previous reports to only build upon protocols that can be integrated into a high throughput strategy consistent with automation. Duplication of these and results from comparable standards could bring an important analytical focus to carbohydrate sequencing that is greatly lacking.

Keywords

Mannose GlcNAc Neutral Loss Trisaccharide Aminop Yrene 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

References

  1. 1.
    Costello, C. E.; Contado-Miller, J. M.; Cipollo, J. F. A Glycomics Platform for the Analysis of Permethylated Oligosaccharide Alditols. J. Am. Soc. Mass Spectrom. 2007, 18, 1799–1812.CrossRefGoogle Scholar
  2. 2.
    Zhao, C.; Xie, B.; Chan, S. Y.; Costello, C. E.; O’Connor, P. B. Collisionally Activated Dissociation and Electron Capture Dissociation Provide Complementary Structural Information for Branched Permethylated Oligosaccharides. J. Am. Soc. Mass Spectrom. 2008, 19, 138–150.CrossRefGoogle Scholar
  3. 3.
    Zhuang, Z.; Starkey, J. A.; Mechref, Y.; Novotny, M. V.; Jacobson, S. C. Electrophoretic Analysis of N-Glycans on Microfluidic Devices. Anal. Chem. 2007, 79, 7170–7175.CrossRefGoogle Scholar
  4. 4.
    Ashline, D. J.; Lapadula, A. J.; Liu, Y. H.; Lin, M.; Grace, M.; Pramanik, B.; Reinhold, V. N. Carbohydrate Structural Isomers Analyzed by Sequential Mass Spectrometry. Anal. Chem. 2007, 79, 3830–3842.CrossRefGoogle Scholar
  5. 5.
    Prien, J. M.; Huysentruyt, L. C.; Ashline, D. J.; Lapadula, A. J.; Seyfried, T. N.; Reinhold, V. N. Differentiating N-Linked Glycan Structural Isomers in Metastatic and Nonmetastatic Tumor Cells Using Sequential Mass Spectrometry. Glycobiology 2008, 18, 353–366.CrossRefGoogle Scholar
  6. 6.
    Zhang, H.; Singh, S.; Reinhold, V. N. Congruent Strategies for Carbohydrate Sequencing, II: FragLib: An MSn Fragment Spectra Fingerprint Library. Anal. Chem. 2005, 77.Google Scholar
  7. 7.
    Lapadula, A. J.; Hatcher, P. J.; Hanneman, A. J.; Ashline, D. J.; Zhang, H.; Reinhold, V. N. Congruent Strategies for Carbohydrate Sequencing: 3. OSCAR: An Algorithm for Assigning Oligosaccharide Topology from MSn data. Anal. Chem. 2005, 77, 6271–6279.CrossRefGoogle Scholar
  8. 8.
    Ciucanu, I.; Kerek, F. A Simple and Rapid Method for the Permethylation of Carbohydrates. Carbohydr. Res. 1984, 131, 209–217.CrossRefGoogle Scholar
  9. 9.
    Fu, D.; Chen, L.; O’Neill, R. A. A Detailed Structural Characterization of Ribonuclease B Oligosaccharides by 1H NMR Spectroscopy and Mass Spectrometry. Carbohydr. Res. 1994, 261, 173–186.CrossRefGoogle Scholar
  10. 10.
    Guttman, A.; Pritchett, T. Capillary Gel Electrophoresis Separation of High-Mannose Type Oligosaccharides Derivatized by l-Aminopyrene-3,6,8-Trisulfonic acid. Electrophoresis 1995, 16, 1906–1911.CrossRefGoogle Scholar
  11. 11.
    Vliegenthart, J. F. G. High Resolution H-1 Nuclear Magnetic Resonance Spectroscopy as a Tool in the Structural Analysis of Carbohydrates Related to Glycoproteins. Adv. Carbohydr. Chem. Biochem. 1983, 41, 209–374.CrossRefGoogle Scholar
  12. 12.
    Kurimoto, A.; Daikoku, S.; Mutsuga, S.; Kanie, O. Analysis of Energy-Resolved Mass Spectra at MSn in a Pursuit to Characterize Structural Isomers of Oligosaccharides. Anal. Chem. 2006, 78, 3461–3466.CrossRefGoogle Scholar
  13. 13.
    Liang, C. J.; Yamashita, K.; Kobata, A. Structural Study of the Carbohydrate Moiety of Bovine Pancreatic Ribonuclease B. J. Biochem. 1980, 88, 51–58.Google Scholar
  14. 14.
    Kakehi, K.; Suzuki, S.; Honda, S.; Lee, Y. C. Precolumn Labeling of Reducing Carbohydrates with 1 -(p-Methoxy)Phenyl-3-Methyl-5-Pyrazolone: Analysis of Neutral and Sialic Acid-Containing Oligosaccharides Found in Glycoproteins. Anal. Biochem. 1991, 199, 256–268.CrossRefGoogle Scholar
  15. 15.
    Laine, R. A. Invited Commentary, Glyco-Forum. Glycobiology 1994, 4, 759–767.CrossRefGoogle Scholar
  16. 16.
    Clowers, B. H.; Dwivedi, P.; Steiner, W. E.; Hill, H. H.; Bendiak, B. Separation of Sodiated Isobaric Disaccharides and Trisaccharides Using Electrospray Ionization-Atmospheric Pressure Ion Mobility-Time of Flight Mass Spectrometry. J. Am. Soc. Mass Spectrom. 2007, 16, 660–669.CrossRefGoogle Scholar
  17. 17.
    Dwivedi, P.; Bendiak, B.; Clowers, B. H.; Hill, H. H. Jr. Rapid Resolution of Carbohydrate Isomers by Electrospray Ionization Ambient Pressure Ion Mobility Spectrometry-Time-of-Flight Mass Spectrometry (ESI-APIMS-TOFMS). J. Am. Soc. Mass Spectrom. 2007, 18, 1163–1175.CrossRefGoogle Scholar

Copyright information

© American Society for Mass Spectrometry 2009

Authors and Affiliations

  • Justin M. Prien
    • 1
  • David J. Ashline
    • 1
  • Anthony J. Lapadula
    • 3
  • Hailong Zhang
    • 1
  • Vernon N. Reinhold
    • 1
    • 2
  1. 1.The Glycomics Center, Division of Molecular, Cellular, and Biomedical SciencesUniversity of New HampshireDurhamUSA
  2. 2.The Glycomics Center, Department of ChemistryUniversity of New HampshireDurhamUSA
  3. 3.The Glycomics Center, Department of Computer ScienceUniversity of New Hampshire, DurhamUSA

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