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Mannose7 glycan isomer characterization by IMS-MS/MS analysis.

Feifei Zhu1, Sunyoung Lee, Stephen J Valentine

  • 1Department of Chemistry, Indiana University, 800 Kirkwood Ave., Bloomington, IN 47405, USA.

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|October 12, 2012
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Summary

Researchers used ion mobility spectrometry-tandem mass spectrometry (IMS-MS/MS) to characterize mannose-7 N-acetylglucosamine-2 (Man(7)GlcNAc(2)) glycan isomers from bovine ribonuclease B. Distinct fragmentation patterns confirmed the presence of four unique glycan isomers.

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Area of Science:

  • Glycomics
  • Analytical Chemistry
  • Biochemistry

Background:

  • Bovine ribonuclease B is a glycoprotein containing N-linked glycans.
  • Characterization of glycan isomers is crucial for understanding their biological roles.
  • Ion mobility spectrometry-tandem mass spectrometry (IMS-MS/MS) offers high-resolution separation and structural analysis of complex carbohydrates.

Purpose of the Study:

  • To characterize the isomers of the Man(7)GlcNAc(2) glycan found in bovine ribonuclease B.
  • To differentiate between various glycan isomers using IMS-MS/MS.
  • To confirm the existence and number of proposed glycan isomers.

Main Methods:

  • Utilizing ion mobility spectrometry (IMS) to separate precursor ions ([Man7 + 2Na](2+)) based on their mobility.
  • Employing tandem mass spectrometry (MS/MS) in an ion trap to fragment selected precursor ions.
  • Analyzing fragmentation spectra and extracted fragment ion drift time distributions (XFIDTDs) for isomer identification.

Main Results:

  • Specific fragmentation spectra were obtained for different precursor ion mobilities, indicating successful isomer isolation or enrichment.
  • A fragment ion at m/z 903.8 was identified, corresponding to the loss of an internal mannose residue from a specific isomer.
  • Distinctive precursor ion drift time profiles from XFIDTDs confirmed the presence of four separate Man(7)GlcNAc(2) glycan isomers.

Conclusions:

  • IMS-MS/MS is effective for characterizing and differentiating glycan isomers.
  • The study confirms the existence of four distinct Man(7)GlcNAc(2) isomers in bovine ribonuclease B.
  • These findings contribute to a deeper understanding of glycan heterogeneity and structural diversity.