Skip to main content
SpringerLink
Log in

Assignment of the Stereochemistry and Anomeric Configuration of Sugars within Oligosaccharides Via Overlapping Disaccharide Ladders Using MSn

  • Research Article
  • Published:
Journal of The American Society for Mass Spectrometry

Abstract

A systematic approach is described that can pinpoint the stereo-structures (sugar identity, anomeric configuration, and location) of individual sugar units within linear oligosaccharides. Using a highly modified mass spectrometer, dissociation of linear oligosaccharides in the gas phase was optimized along multiple-stage tandem dissociation pathways (MSn, n = 4 or 5). The instrument was a hybrid triple quadrupole/linear ion trap mass spectrometer capable of high-efficiency bidirectional ion transfer between quadrupole arrays. Different types of collision-induced dissociation (CID), either on-resonance ion trap or beam-type CID could be utilized at any given stage of dissociation, enabling either glycosidic bond cleavages or cross-ring cleavages to be maximized when wanted. The approach first involves optimizing the isolation of disaccharide units as an ordered set of overlapping substructures via glycosidic bond cleavages during early stages of MSn, with explicit intent to minimize cross-ring cleavages. Subsequently, cross-ring cleavages were optimized for individual disaccharides to yield key diagnostic product ions (m/z 221). Finally, fingerprint patterns that establish stereochemistry and anomeric configuration were obtained from the diagnostic ions via CID. Model linear oligosaccharides were derivatized at the reducing end, allowing overlapping ladders of disaccharides to be isolated from MSn. High confidence stereo-structural determination was achieved by matching MSn CID of the diagnostic ions to synthetic standards via a spectral matching algorithm. Using this MSn (n = 4 or 5) approach, the stereo-structures, anomeric configurations, and locations of three individual sugar units within two pentasaccharides were successfully determined.

ᅟ

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

Scheme 1
Figure 1
Figure 2
Figure 3

Similar content being viewed by others

References

  1. Apweiler, R., Hermjakob, H., Sharon, N.: On the frequency of protein glycosylation, as deduced from analysis of the SWISS-PROT database. Biochim. Biophys. Acta 1473, 4–8 (1999)

    Article  CAS  Google Scholar 

  2. Eichler, J., Michael, W.W.A.: Post-translational protein modification in Archaea. Microbiol. Mol. Biol. Rev. 69, 393–425 (2005)

    Article  CAS  Google Scholar 

  3. Mescher, M.F., Strominger, J.L.: Purification and characterization of a prokaryotic glucoprotein from the cell envelope of Halobacterium salinarium. J. Biol. Chem. 251, 2005–2014 (1976)

    Google Scholar 

  4. Abu-Qarn, M., Eichler, J., Sharon, N.: Not just for Eukarya anymore: protein glycosylation in bacteria and archaea. Curr. Opin. Struct. Biol. 18, 544–550 (2008)

    Article  CAS  Google Scholar 

  5. Perkel, J.M.: Life science technologies: Glycoproteomics: the sweet smell of we’re-getting-there. Science 331, 95–97 (2011)

    Article  Google Scholar 

  6. Geyer, H., Geyer, R.: Strategies for analysis of glycoprotein glycosylation. Biochim. Biophys. Acta Gen. Subj. 1764, 1853–1869 (2006)

    CAS  Google Scholar 

  7. Duus, J., Gotfredsen, C.H., Bock, K.: Carbohydrate structural determination by NMR spectroscopy: modern methods and limitations. Chem. Rev. 100, 4589–4614 (2000)

    Article  CAS  Google Scholar 

  8. Fellenberg, M., Coksezen, A., Meyer, B.: Characterization of picomole amounts of oligosaccharides from glycoproteins by 1H NMR spectroscopy. Angew. Chem. Int. Ed. 49, 2630–2633 (2010)

    Article  CAS  Google Scholar 

  9. Tseng, K., Hedrick, J.L., Lebrilla, C.B.: Catalog-library approach for the rapid and sensitive structural elucidation of oligosaccharides. Anal. Chem. 71, 3747–3754 (1999)

    Article  CAS  Google Scholar 

  10. Zaia, J.: Mass spectrometry of oligosaccharides. Mass Spectrom. Rev. 23, 161–227 (2004)

    Article  CAS  Google Scholar 

  11. Reinhold, V.N., Reinhold, B.B., Costello, C.E.: Carbohydrate molecular weight profiling, sequence, linkage, and branching data: ES-MS and CID. Anal. Chem. 67, 1772–1784 (1995)

    Article  CAS  Google Scholar 

  12. Peltier, J.M., Maclean, D.B., Szarek, W.A.: Determination of the glycosidic linkage in peracetylated disaccharides comprised of D-glucopyranose units by use of desorption electron-ionization mass-spectrometry. Rapid Commun. Mass Spectrom. 5, 446–449 (1991)

    Article  CAS  Google Scholar 

  13. Asam, M.R., Glish, G.L.: Tandem mass spectrometry of alkali cationized polysaccharides in a quadrupole ion trap. J. Am. Soc. Mass Spectrom. 8, 987–995 (1997)

    Article  CAS  Google Scholar 

  14. Polfer, N.C., Valle, J.J., Moore, D.T., Oomens, J., Eyler, J.R., Bendiak, B.: Differentiation of isomers by wavelength-tunable infrared multiple-photon dissociation-mass spectrometry: application to glucose-containing disaccharides. Anal. Chem. 78, 670–679 (2006)

    Article  CAS  Google Scholar 

  15. Dongré, A.R., Wysocki, V.H.: Linkage position determination of lithium cationized disaccharides by surface-induced dissociation tandem mass spectrometry. Org. Mass Spectrom. 29, 700–702 (1994)

    Article  Google Scholar 

  16. Domon, B., Müller, D.R., Richter, W.J.: High performance tandem mass spectrometry for sequence, branching and interglycosidic linkage analysis of peracetylated oligosaccharides. Biol. Mass Spectrom. 19, 390–392 (1990)

    Article  CAS  Google Scholar 

  17. Sheeley, D.M., Reinhold, V.N.: Structural characterization of carbohydrate sequence, linkage, and branching in a quadrupole ion trap mass spectrometer: neutral oligosaccharides and N-linked glycans. Anal. Chem. 70, 3053–3059 (1998)

    Article  CAS  Google Scholar 

  18. Chai, W., Lawson, A., Piskarev, V.: Branching pattern and sequence analysis of underivatized oligosaccharides by combined MS/MS of singly and doubly charged molecular ions in negative-ion electrospray mass spectrometry. J. Am. Soc. Mass Spectrom. 13, 670–679 (2002)

    Article  Google Scholar 

  19. Hofmeister, G.E., Zhou, Z., Leary, J.A.: Linkage position determination in lithium-cationized disaccharides: tandem mass spectrometry and semiempirical calculations. J. Am. Chem. Soc. 113, 5964–5970 (1991)

    Article  CAS  Google Scholar 

  20. Xie, Y., Lebrilla, C.B.: Infrared multiphoton dissociation of alkali metal-coordinated oligosaccharides. Anal. Chem. 75, 1590–1598 (2003)

    Article  CAS  Google Scholar 

  21. Reilly, J.P.: Ultraviolet photofragmentation of biomolecular ions. Mass Spectrom. Rev. 28, 425–447 (2009)

    Article  CAS  Google Scholar 

  22. Adamson, J.T., Håkansson, K.: Electron capture dissociation of oligosaccharides ionized with alkali, alkaline earth, and transition metals. Anal. Chem. 79, 2901–2910 (2007)

    Article  CAS  Google Scholar 

  23. Zhao, C., Xie, B., Chan, S.-Y., Costello, C., O’Connor, P.: Collisionally activated dissociation and electron capture dissociation provide complementary structural information for branched permethylated oligosaccharides. J. Am. Soc. Mass Spectrom. 19, 138–150 (2008)

    Article  CAS  Google Scholar 

  24. Wolff, J., Laremore, T., Aslam, H., Linhardt, R., Amster, I.J.: Electron-induced dissociation of glycosaminoglycan tetrasaccharides. J. Am. Soc. Mass Spectrom. 19, 1449–1458 (2008)

    Article  CAS  Google Scholar 

  25. Wolff, J., Amster, I.J., Chi, L., Linhardt, R.: Electron detachment dissociation of glycosaminoglycan tetrasaccharides. J. Am. Soc. Mass Spectrom. 18, 234–244 (2007)

    Article  CAS  Google Scholar 

  26. Yu, X., Huang, Y., Lin, C., Costello, C.E.: Energy-dependent electron activated dissociation of metal-adducted permethylated oligosaccharides. Anal. Chem. 84, 7487–7494 (2012)

    Article  CAS  Google Scholar 

  27. Gennaro, L.A., Harvey, D.J., Vouros, P.: Reversed-phase ion-pairing liquid chromatography/ion trap mass spectrometry for the analysis of negatively charged, derivatized glycans. Rapid Commun. Mass Spectrom. 17, 1528–1534 (2003)

    Article  CAS  Google Scholar 

  28. Domon, B., Mueller, D.R., Richter, W.J.: Tandem mass-spectrometric analysis of fixed-charge derivatized oligosaccharides. Org. Mass Spectrom. 29, 713–719 (1994)

    Article  CAS  Google Scholar 

  29. Bendiak, B., Fang, T.T.: Assignment of the stereochemistry and anomeric configuration of structurally informative product ions derived from disaccharides: infrared photodissociation of glycosyl-glycolaldehydes in the negative ion mode. Carbohydr. Res. 345, 2390–2400 (2010)

    Article  CAS  Google Scholar 

  30. Laine, R.A.: A calculation of all possible oligosaccharide isomers both branched and linear yields 1.05 × 1012 structures for a reducing hexasaccharide - the isomer-barrier to development of single-method saccharide sequencing or synthesis systems. Glycobiology 4, 759–767 (1994)

    Article  CAS  Google Scholar 

  31. Gaucher, S.P., Leary, J.A.: Determining anomericity of the glycosidic bond in Zn(II)-diethylenetriamine–disaccharide complexes using MSn in a quadrupole ion trap. J. Am. Soc. Mass Spectrom. 10, 269–272 (1999)

    Article  CAS  Google Scholar 

  32. Smith, G., Leary, J.A.: Differentiation of stereochemistry of glycosidic bond configuration: tandem mass spectrometry of diastereomeric cobalt-glucosyl-glucose disaccharide complexes. J. Am. Soc. Mass Spectrom. 7, 953–957 (1996)

    Article  CAS  Google Scholar 

  33. Mulroney, B., Traeger, J.C., Stone, B.A.: Determination of both linkage position and anomeric configuration in underivatized glucopyranosyl disaccharides by electrospray mass spectrometry. J. Mass Spectrom. 30, 1277–1283 (1995)

    Article  CAS  Google Scholar 

  34. Guan, B., Cole, R.B.: MALDI linear-field reflectron TOF post-source decay analysis of underivatized oligosaccharides: determination of glycosidic linkages and anomeric configurations using anion attachment. J. Am. Soc. Mass Spectrom. 19, 1119–1131 (2008)

    Article  CAS  Google Scholar 

  35. Simoes, J., Domingues, P., Reis, A., Nunes, F.M., Coimbra, M.A., Domingues, R.M.: Identification of anomeric configuration of underivatized reducing glucopyranosyl-glucose Disaccharides by tandem mass Spectrometry and Multivariate analysis. Anal. Chem. 79, 5896–5905 (2007)

    Article  CAS  Google Scholar 

  36. Gaucher, S.P., Leary, J.A.: Stereochemical differentiation of mannose, glucose, galactose, and talose using zinc(II) diethylenetriamine and ESI-ion trap mass spectrometry. Anal. Chem. 70, 3009–3014 (1998)

    Article  CAS  Google Scholar 

  37. Salpin, J.Y., Tortajada, J.: Structural characterization of hexoses and pentoses using lead cationization. An electrospray ionization and tandem mass spectrometric study. J. Mass Spectrom. 37, 379–388 (2002)

    Article  CAS  Google Scholar 

  38. Zhu, X.Y., Sato, T.: The distinction of underivatized monosaccharides using electrospray ionization ion trap mass spectrometry. Rapid Commun. Mass Spectrom. 21, 191–198 (2007)

    Article  CAS  Google Scholar 

  39. Brown, D.J., Stefan, S.E., Berden, G., Steill, J.D., Oomens, J., Eyler, J.R., Bendiak, B.: Direct evidence for the ring opening of monosaccharide anions in the gas phase: photodissociation of aldohexoses and aldohexoses derived from disaccharides using variable-wavelength infrared irradiation in the carbonyl stretch region. Carbohydr. Res. 346, 2469–2481 (2011)

    Article  CAS  Google Scholar 

  40. Augusti, D.V., Carazza, F., Augusti, R., Tao, W.A., Cooks, R.G.: Quantitative chiral analysis of sugars by electrospray ionization tandem mass spectrometry using modified amino acids as chiral reference compounds. Anal. Chem. 74, 3458–3462 (2002)

    Article  CAS  Google Scholar 

  41. Ashline, D., Singh, S., Hanneman, A., Reinhold, V.: Congruent strategies for carbohydrate sequencing. 1. Mining structural details by MSn. Anal. Chem. 77, 6250–6262 (2005)

    Article  CAS  Google Scholar 

  42. 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. 79, 3830–3842 (2007)

    Article  CAS  Google Scholar 

  43. Fang, T.T., Bendiak, B.: The stereochemical dependence of unimolecular dissociation of monosaccharide-glycolaldehyde anions in the gas phase: a basis for assignment of the stereochemistry and anomeric configuration of monosaccharides in oligosaccharides by mass spectrometry via a key discriminatory product ion of disaccharide fragmentation, m/z 221. J. Am. Chem. Soc. 129, 9721–9736 (2007)

    Article  CAS  Google Scholar 

  44. Viseux, N., de Hoffmann, E., Domon, B.: Structural assignment of permethylated oligosaccharide subunits using sequential tandem mass spectrometry. Anal. Chem. 70, 4951–4959 (1998)

    Article  CAS  Google Scholar 

  45. Garozzo, D., Giuffrida, M., Impallomeni, G., Ballistreri, A., Montaudo, G.: Determination of linkage position and identification of the reducing end in linear oligosaccharides by negative ion fast atom bombardment mass spectrometry. Anal. Chem. 62, 279–286 (1990)

    Article  CAS  Google Scholar 

  46. Dallinga, J.W., Heerma, W.: Reaction mechanism and fragment ion structure determination of deprotonated small oligosaccharides, studied by negative ion fast atom bombardment (tandem) mass spectrometry. Biol. Mass Spectrom. 20, 215–231 (1991)

    Article  CAS  Google Scholar 

  47. Carroll, J.A., Willard, D., Lebrilla, C.B.: Energetics of cross-ring cleavages and their relevance to the linkage determination of oligosaccharides. Anal. Chim. Acta. 307, 431–447 (1995)

    Article  CAS  Google Scholar 

  48. Fang, T.T., Zirrolli, J., Bendiak, B.: Differentiation of the anomeric configuration and ring form of glucosyl-glycolaldehyde anions in the gas phase by mass spectrometry: isomeric discrimination between m/z 221 anions derived from disaccharides and chemical synthesis of m/z 221 standards. Carbohydr. Res. 342, 217–235 (2007)

    Article  CAS  Google Scholar 

  49. Konda, C., Bendiak, B., Xia, Y.: Differentiation of the stereochemistry and anomeric configuration for 1-3 linked disaccharides via tandem mass spectrometry and 18O-labeling. J. Am. Soc. Mass Spectrom. 23, 347–358 (2012)

    Article  CAS  Google Scholar 

  50. Harvey, D.J.: Electrospray mass spectrometry and fragmentation of N-linked carbohydrates derivatized at the reducing terminus. J. Am. Soc. Mass Spectrom. 11, 900–915 (2000)

    Article  CAS  Google Scholar 

  51. Londry, F.A., Hager, J.: Mass selective axial ion ejection from a linear quadrupole ion trap. J. Am. Soc. Mass Spectrom. 14, 1130–1147 (2003)

    Article  CAS  Google Scholar 

  52. Zhang, Z.: Prediction of low-energy collision-induced dissociation spectra of peptides. Anal. Chem. 76, 3908–3922 (2004)

    Article  CAS  Google Scholar 

  53. Domon, B., Costello, C.E.: A systematic nomenclature for carbohydrate fragmentations in FAB-MS/MS spectra of glycoconjugates. Glycoconj. J. 5, 397–409 (1988)

    Article  CAS  Google Scholar 

  54. Thomson, B.A.: Apparatus and method for MSnth in a tandem mass spectrometer system. U.S. Patent 7 145, 133–B142 (2006)

    Google Scholar 

  55. Xia, Y., Thomson, B.A., McLuckey, S.A.: Bidirectional ion transfer between quadrupole arrays: MSn ion/ion reaction experiments on a quadrupole/time-of-flight tandem mass spectrometer. Anal. Chem. 79, 8199–8206 (2007)

    Article  CAS  Google Scholar 

  56. Thomson, B.A., Jolliffe, C.L.: Spectrometer with axial field. U.S. Patent 5 847, 386 (1998)

    Google Scholar 

  57. Loboda, A., Krutchinsky, A., Loboda, O., McNabb, J., Spicer, V., Ens, W., Standing, K.: Novel linad II electrode geometry for creating an axial field in a mulitpole ion guide. Eur. J. Mass Spectrom. 6, 531–536 (2000)

    Article  CAS  Google Scholar 

  58. Collings, B.A.: Fragmentation of ions in a low pressure linear ion trap. J. Am. Soc. Mass Spectrom. 18, 1459–1466 (2007)

    Article  CAS  Google Scholar 

  59. Morris, M., Pierre, T., Boyd, R.K.: Characterization of a high-pressure quadrupole collision cell for low-energy collision-induced dissociation. J. Am. Soc. Mass Spectrom. 5, 1042–1063 (1994)

    Article  CAS  Google Scholar 

Download references

Acknowledgements

The authors thank Y. Du and Professor Z. Ouyang for conducting spectral similarity score calculations and Dr. L. J. Campbell for helpful discussions. C.K. and Y.X. thank the support from Purdue Research Foundation. B.B. acknowledges financial support from NSF CHE-0137986 and CHE-0718007.

Author information

Authors and Affiliations

  1. Department of Chemistry, Purdue University, West Lafayette, IN, 47907, USA

    Chiharu Konda & Yu Xia

  2. AB SCIEX, Concord, ON, L4K 4V8, Canada

    Frank A. Londry

  3. Department of Cell and Developmental Biology, and Program in Structural Biology and Biophysics, University of Colorado-Denver, Anschutz Medical Campus, Aurora, CO, 80045, USA

    Brad Bendiak

Authors
  1. Chiharu Konda
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Frank A. Londry
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Brad Bendiak
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Yu Xia
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Yu Xia.

Electronic supplementary material

Below is the link to the electronic supplementary material.

ESM 1

(PDF 716 kb)

Rights and permissions

Reprints and permissions

About this article

Cite this article

Konda, C., Londry, F.A., Bendiak, B. et al. Assignment of the Stereochemistry and Anomeric Configuration of Sugars within Oligosaccharides Via Overlapping Disaccharide Ladders Using MSn . J. Am. Soc. Mass Spectrom. 25, 1441–1450 (2014). https://doi.org/10.1007/s13361-014-0881-4

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s13361-014-0881-4

Key words

Search

Navigation