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Distinctive characteristics of MALDI-Q/TOF and TOF/TOF tandem mass spectrometry for sequencing of permethylated complex type N-glycans

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Abstract

Concerted MALDI-MS profiling and CID MS/MS sequencing of permethylated glycans is one of the most effective approaches for high throughput glycomics applications. In essence, the identification of larger complex type N-glycans necessitates an unambiguous definition of any modification on the trimannosyl core and the complement of non-reducing terminal sequences which constitute the respective antennary structures. Permethylation not only affords analyses of both neutral and sialylated glycans at comparable ease and sensitivity but also yields more sequence-informative fragmentation pattern. Facile glycosidic cleavages directed mostly at N-acetylglucosamine under low energy CID, as implemented on a quadrupole/time-of-flight (Q/TOF) instrument, often afford multiple losses of the attached antenna resulting in characteristic ions related to the number of antennary branches on the trimannosyl core. Non-reducing terminal epitopes can be easily deduced but information on the linkage specific substituent on the terminal units is often missing. The high energy CID MS/MS afforded by TOF/TOF instrument can fill in the gap by giving an array of additional cross-ring and satellite ions. Glycosidic cleavages occurring specifically in concert with loss of 2-linked or 3-linked substituents provide an effective way to identify the branch-specific antennary extension. These characteristics are shown here to be effective in deriving the sequences of additionally galactosylated, sialylated and fucosylated terminal N-acetyllactosamine units and their antennary location. Together, a highly reproducible fragmentation pattern can be formulated to simplify spectral assignment. This work also provides first real examples of sequencing multiply sialylated complex type N-glycans by high energy CID on a TOF/TOF instrument.

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Abbreviations

CID:

collision-induced dissociation

EI:

electron impact

ESI:

electrospray ionization

FAB:

fast atom bombardment

MALDI:

matrix assisted laser desorption/ionization

MS:

mass spectrometry

MS/MS:

tandem mass spectrometry

Q/TOF:

quadrupole/time-of-flight

LacNAc:

N-aectyllactosamine

OMe:

O-methyl substituent or methoxy

References

  1. Sutton-Smith, M., Morris, H.R., Grewal, P.K., Hewitt, J.E., Bittner, R.E., Goldin, E., Schiffmann, R., Dell, A.: MS screening strategies: Investigating the glycomes of knockout and myodystrophic mice and leukodystrophic human brains. Biochem. Soc. Symp. 69, 105–15 (2002)

    PubMed  CAS  Google Scholar 

  2. Kochetkov, N.K., Chizhov, O.S.: Mass spectrometry of carbohydrate derivatives. Adv. Carbohydr. Chem. Biochem. 21, 39–93 (1966)

    PubMed  CAS  Google Scholar 

  3. Egge, H., Peter-Katalinic, J.: Fast atom bombardment mass spectrometry for structural elucidation of glycoconjugates. Mass Spectrom. Rev. 6, 331–93 (1987)

    Article  CAS  Google Scholar 

  4. Dell, A.: F.A.B.-mass spectrometry of carbohydrates. Adv. Carbohydr. Chem. Biochem. 45, 19–72 (1987)

    Article  PubMed  CAS  Google Scholar 

  5. Lemoine, J., Fournet, B., Despeyroux, D., Jennings, K.R., Rosenberg, R., de Hoffmann, E.: Collision-induced dissociation of alkali metal cationized and permethylated oligosaccharides: Influence of the collision energy and of the collision gas for the assignment of linkage position. J. Am. Soc. Mass Spectrom. 4(3), 197–302 (1993)

    Article  CAS  Google Scholar 

  6. Reinhold, V.N., Reinhold, B.B., Chan, S.: Carbohydrate sequence analysis by electrospray ionization-mass spectrometry. Methods Enzymol. 271, 377–402 (1996)

    Article  PubMed  CAS  Google Scholar 

  7. Weiskopf, A.S., Vouros, P., Harvey, D.J.: Characterization of oligosaccharide composition and structure by quadrupole ion trap mass spectrometry. Rapid Commun. Mass Spectrom. 11(14), 1493–504 (1997)

    Article  PubMed  CAS  Google Scholar 

  8. Viseux, N., de Hoffmann, E., Domon, B.: Structural analysis of permethylated oligosaccharides by electrospray tandem mass spectrometry. Anal. Chem. 69(16), 3193–98 (1997)

    Article  PubMed  CAS  Google Scholar 

  9. Weiskopf, A.S., Vouros, P., Harvey, D.J.: Electrospray ionization-ion trap mass spectrometry for structural analysis of complex N-linked glycoprotein oligosaccharides. Anal. Chem. 70(20), 4441–47 (1998)

    Article  PubMed  CAS  Google Scholar 

  10. Viseux, N., de Hoffmann, E., Domon, B.: Structural assignment of permethylated oligosaccharide subunits using sequential tandem mass spectrometry. Anal. Chem. 70(23), 4951–59 (1998)

    Article  PubMed  CAS  Google Scholar 

  11. Morelle, W., Faid, V., Michalski, J.C.: Structural analysis of permethylated oligosaccharides using electrospray ionization quadrupole time-of-flight tandem mass spectrometry and deutero-reduction. Rapid Commun. Mass Spectrom. 18(20), 2451 (2004)

    Article  PubMed  CAS  Google Scholar 

  12. Ashline, D., Singh, S., Hanneman, A., Reinhold, V.: Congruent Strategies for Carbohydrate Sequencing. 1. Mining Structural Details by MS(n). Anal. Chem. 77(19), 6250–62 (2005)

    Article  PubMed  CAS  Google Scholar 

  13. Harvey, D.J.: Collision-induced fragmentation of underivatized N-linked carbohydrates ionized by electrospray. J. Mass Spectrom. 35(10), 1178–90 (2000)

    Article  PubMed  CAS  Google Scholar 

  14. Chai, W., Piskarev, V., Lawson, A.M.: Negative-ion electrospray mass spectrometry of neutral underivatized oligosaccharides. Anal. Chem. 73(3), 651–57 (2001)

    Article  PubMed  CAS  Google Scholar 

  15. Sagi, D., Peter-Katalinic, J., Conradt, H.S., Nimtz, M.: Sequencing of tri- and tetraantennary N-glycans containing sialic acid by negative mode ESI QTOF tandem MS. J. Am. Soc. Mass Spectrom. 13(9), 1138–48 (2002)

    Article  PubMed  CAS  Google Scholar 

  16. Chai, W., Piskarev, V., Lawson, A.M.: 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(6), 670–79 (2002)

    Article  PubMed  Google Scholar 

  17. Robbe, C., Capon, C., Coddeville, B., Michalski, J.C.: Diagnostic ions for the rapid analysis by nano-electrospray ionization quadrupole time-of-flight mass spectrometry of O-glycans from human mucins. Rapid Commun. Mass Spectrom. 18(4), 412–20 (2004)

    Article  PubMed  CAS  Google Scholar 

  18. Harvey, D.J.: Fragmentation of negative ions from carbohydrates: part 1. Use of nitrate and other anionic adducts for the production of negative ion electrospray spectra from N-linked carbohydrates. J. Am. Soc. Mass Spectrom. 16(5), 622–30 (2005)

    Article  PubMed  CAS  Google Scholar 

  19. Harvey, D.J.: Fragmentation of negative ions from carbohydrates: part 2. Fragmentation of high-mannose N-linked glycans. J. Am. Soc. Mass Spectrom. 16(5), 631–46 (2005)

    Article  PubMed  CAS  Google Scholar 

  20. Harvey, D.J.: Fragmentation of negative ions from carbohydrates: Part 3. Fragmentation of hybrid and complex N-linked glycans. J. Am. Soc. Mass Spectrom. 16(5), 647–59 (2005)

    Article  PubMed  CAS  Google Scholar 

  21. Harvey, D.J.: N-(2-diethylamino)ethyl-4-aminobenzamide derivative for high sensitivity mass spectrometric detection and structure determination of N-linked carbohydrates. Rapid Commun. Mass Spectrom. 14(10), 862–71 (2000)

    Article  PubMed  CAS  Google Scholar 

  22. Harvey, D.J.: Electrospray mass spectrometry and fragmentation of N-linked carbohydrates derivatized at the reducing terminus. J. Am. Soc. Mass Spectrom. 11(10), 900–15 (2000)

    Article  PubMed  CAS  Google Scholar 

  23. Lattova, E., Perreault, H.: Profiling of N-linked oligosaccharides using phenylhydrazine derivatization and mass spectrometry. J. Chromatogr. A 1016(1), 71–87 (2003)

    Article  PubMed  CAS  Google Scholar 

  24. Takegawa, Y., Ito, S., Yoshioka, S., Deguchi, K., Nakagawa, H., Monde, K., Nishimura, S.I.: Structural assignment of isomeric 2-aminopyridine-derivatized oligosaccharides using MSn spectral matching. Rapid Commun. Mass Spectrom. 18(4), 385–91 (2004)

    Article  PubMed  CAS  Google Scholar 

  25. Morelle, W., Michalski, J.C.: Sequencing of oligosaccharides derivatized with benzylamine using electrospray ionization-quadrupole time of flight-tandem mass spectrometry. Electrophoresis 25(14), 2144–55 (2004)

    Article  PubMed  CAS  Google Scholar 

  26. Morelle, W., Page, A., Michalski, J.C.: Electrospray ionization ion trap mass spectrometry for structural characterization of oligosaccharides derivatized with 2-aminobenzamide. Rapid Commun. Mass Spectrom. 19(9), 1145–58 (2005)

    Article  PubMed  CAS  Google Scholar 

  27. Harvey, D.J.: Collision-induced fragmentation of negative ions from N-linked glycans derivatized with 2-aminobenzoic acid. J. Mass Spectrom. 40(5), 642–53 (2005)

    Article  PubMed  CAS  Google Scholar 

  28. Kameyama, A., Kikuchi, N., Nakaya, S., Ito, H., Sato, T., Shikanai, T., Takahashi, Y., Takahashi, K., Narimatsu, H.: A Strategy for identification of oligosaccharide structures using observational multistage mass spectral library. Anal. Chem. 77(15), 4719–25 (2005)

    Article  PubMed  CAS  Google Scholar 

  29. Dell, A., Morris, H.R.: Glycoprotein structure determination by mass spectrometry. Science 291(5512), 2351–56 (2001)

    Article  PubMed  CAS  Google Scholar 

  30. Harvey, D.J., Bateman, R.H., Bordoli, R.S., Tyldesley, R.: Ionisation and fragmentation of complex glycans with a quadrupole time-of-flight mass spectrometer fitted with a matrix-assisted laser desorption/ionisation ion source. Rapid Commun. Mass Spectrom. 14(22), 2135–42 (2000)

    Article  PubMed  CAS  Google Scholar 

  31. Loboda, A.V., Krutchinsky, A.N., Bromirski, M., 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. 14(12), 1047–57 (2000)

    Article  PubMed  CAS  Google Scholar 

  32. Medzihradszky, K.F., Campbell, J.M., Baldwin, M.A., Falick, A.M., Juhasz, P., Vestal, M.L., Burlingame, A.L.: The characteristics of peptide collision-induced dissociation using a high-performance MALDI-TOF/TOF tandem mass spectrometer. Anal. Chem. 72(3), 552–58 (2000)

    Article  PubMed  CAS  Google Scholar 

  33. Harvey, D.J., Postsource decay fragmentation of N-linked carbohydrates from ovalbumin and related glycoproteins. J. Am. Soc. Mass Spectrom. 11(6), 572–77 (2000)

    Article  PubMed  CAS  Google Scholar 

  34. Naven, T.J., Harvey, D.J., Brown, J., Critchley, G.: Fragmentation of complex carbohydrates following ionization by matrix-assisted laser desorption with an instrument fitted with time-lag focusing. Rapid Commun. Mass Spectrom. 11(15), 1681–86 (1997)

    Article  PubMed  CAS  Google Scholar 

  35. Hanrahan, S., Charlwood, J., Tyldesley, R., Langridge, J., Bordoli, R., Bateman, R., Camilleri, P.: Facile sequencing of oligosaccharides by matrix-assisted laser desorption/ionisation on a hybrid quadrupole orthogonal acceleration time-of-flight mass spectrometer. Rapid Commun. Mass Spectrom. 15(14), 1141–51 (2001)

    Article  PubMed  CAS  Google Scholar 

  36. Suzuki, N., Khoo, K.H., Chen, C.M., Chen, H.C., Lee, Y.C.: N-glycan structures of pigeon IgG: A major serum glycoprotein containing Galalpha1–4 Gal termini. J. Biol. Chem. 278(47), 46293–306 (2003)

    Article  PubMed  CAS  Google Scholar 

  37. Terada, M., Khoo, K.H., Inoue, R., Chen, C.I., Yamada, K., Sakaguchi, H., Kadowaki, N., Ma, B.Y., Oka, S., Kawasaki, T., Kawasaki, N.: Characterization of oligosaccharide ligands expressed on SW1116 cells recognized by mannan-binding protein. A highly fucosylated polylactosamine type N-glycan. J. Biol. Chem. 280(12), 10897–913 (2005)

    Article  PubMed  CAS  Google Scholar 

  38. Kui Wong, N., Easton, R.L., Panico, M., Sutton-Smith, M., Morrison, J.C., Lattanzio, F.A., Morris, H.R., Clark, G.F., Dell, A., Patankar, M.S.: Characterization of the oligosaccharides associated with the human ovarian tumor marker CA125. J. Biol. Chem. 278(31), 28619–634 (2003)

    Article  PubMed  CAS  Google Scholar 

  39. Zhang, H., Singh, S., Reinhold, V.N.: Congruent strategies for carbohydrate sequencing. 2. FragLib: An MS(n) spectral library. Anal. Chem. 77(19), 6263–70 (2005)

    Article  PubMed  CAS  Google Scholar 

  40. 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 MS(n) data. Anal. Chem. 77(19), 6271–79 (2005)

    Article  PubMed  CAS  Google Scholar 

  41. 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. 32(2), 167–87 (1997)

    Article  PubMed  CAS  Google Scholar 

  42. Mechref, Y., Novotny, M.V., Krishnan, C.: Structural characterization of oligosaccharides using MALDI-TOF/TOF tandem mass spectrometry. Anal. Chem. 75(18), 4895–903 (2003)

    Article  PubMed  CAS  Google Scholar 

  43. Spina, E., Sturiale, L., Romeo, D., Impallomeni, G., Garozzo, D., Waidelich, D., Glueckmann, M.: New fragmentation mechanisms in matrix-assisted laser desorption/ionization time-of-flight/time-of-flight tandem mass spectrometry of carbohydrates. Rapid Commun. Mass Spectrom. 18(4), 392–98 (2004)

    Article  PubMed  CAS  Google Scholar 

  44. Morelle, W., Slomianny, M.C., Diemer, H., Schaeffer, C., Dorsselaer, A.V., Michalski, J.C.: Fragmentation characteristics of permethylated oligosaccharides using a matrix-assisted laser desorption/ionization two-stage time-of-flight (TOF/TOF) tandem mass spectrometer. Rapid Commun. Mass Spectrom. 18(22), 2637–49 (2004)

    Article  PubMed  CAS  Google Scholar 

  45. Stephens, E., Maslen, S.L., Green, L.G., Williams, D.H.: Fragmentation characteristics of neutral N-linked glycans using a MALDI-TOF/TOF tandem mass spectrometer. Anal. Chem. 76(8), 2343–54 (2004)

    Article  PubMed  CAS  Google Scholar 

  46. Guerardel, Y., Chang, L.Y., Maes, E., Huang, C.J., Khoo, K.H.: Glycomic survey mapping of zebrafish identifies unique sialylation pattern. Glycobiology, in press (2006)

  47. Ciucanu, I., Kerek, F.: A simple and rapid method for the permethylation of carbohydrates. Carbohydr Res. 131(2), 209–17 (1984)

    Article  CAS  Google Scholar 

  48. Dell, A., Reason, A.J., Khoo, K.H., Panico, M., McDowell, R.A., Morris, H.R.: Mass spectrometry of carbohydrate-containing biopolymers. Methods Enzymol. 230, 108–32 (1994)

    PubMed  CAS  Google Scholar 

  49. 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 

  50. Harvey, D.J.: Structural determination of N-linked glycans by matrix-assisted laser desorption/ionization and electrospray ionization mass spectrometry. Proteomics 5(7), 1774–86 (2005)

    Article  PubMed  CAS  Google Scholar 

  51. Cumming, D.A., Hellerqvist, C.G., Harris-Brandts, M., Michnick, S.W., Carver, J.P., Bendiak, B.: Structures of asparagine-linked oligosaccharides of the glycoprotein fetuin having sialic acid linked to N-acetylglucosamine. Biochemistry 28(15), 6500–12 (1989)

    Article  PubMed  CAS  Google Scholar 

  52. Townsend, R.R., Hardy, M.R., Cumming, D.A., Carver, J.P., Bendiak, B.: Separation of branched sialylated oligosaccharides using high-pH anion-exchange chromatography with pulsed amperometric detection. Anal. Biochem. 182(1), 1–8 (1989)

    Article  PubMed  CAS  Google Scholar 

  53. Taguchi, T., Seko, A., Kitajima, K., Muto, Y., Inoue, S., Khoo, K.H., Morris, H.R., Dell, A., Inoue, Y.: Structural studies of a novel type of pentaantennary large glycan unit in the fertilization-associated carbohydrate-rich glycopeptide isolated from the fertilized eggs of Oryzias latipes. J. Biol. Chem. 269(12), 8762–71 (1994)

    PubMed  CAS  Google Scholar 

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Authors and Affiliations

  1. Institute of Biological Chemistry, Academia Sinica, Taiwan

    Shin-Yi Yu, Sz-Wei Wu & Kay-Hooi Khoo

  2. National Core Facilities for Proteomics Research, Academia Sinica, Taiwan

    Kay-Hooi Khoo

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  1. Shin-Yi Yu
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  2. Sz-Wei Wu
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Correspondence to Kay-Hooi Khoo.

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Shin-Yi Yu and Sz-Wei Wu contributed equally to this work.

Dedicated to the late Prof. Yasuo Inoue, without whom the body of work represented by this article would not have been initiated in Taiwan.

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Yu, SY., Wu, SW. & Khoo, KH. Distinctive characteristics of MALDI-Q/TOF and TOF/TOF tandem mass spectrometry for sequencing of permethylated complex type N-glycans. Glycoconj J 23, 355–369 (2006). https://doi.org/10.1007/s10719-006-8492-3

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  • DOI: https://doi.org/10.1007/s10719-006-8492-3

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