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Detection of sialylated N-Linked glycans by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry

  • Biology
  • Published:
Wuhan University Journal of Natural Sciences

Abstract

Native and methyl-esterified sialylated glycans were analyzed with 2, 4, 6-trihydroxyacetophenone (THAP) and 2, 5-dihydroxybenzoic acid (DHB) as matrix by a matrix-assisted laser desorption/ionization time-of-flight mass spectrometer (MALDI-TOF MS). High quality negative-ion spectra of commercial sialylated glycan were obtained with THAP as matrix. Detection limit of the glycan was less than 0.1 pmol. After methyl esterification of sialic acid (SA) residue, sialylated glycans were detected sensitively in the positive-ion mode using DHB as matrix. Neutral and sialylated glycans from the mixture of asialofetuin and fetuin were methylesterified and simultaneously recognized in one manipulation. Methyl esterification of SA residue offers a convenient and sensitive way to identify the structure of N-linked glycans for glycan profiling.

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References

  1. Bork K, Horstkorte R, Weidemann W. Increasing the sialylation of therapeutic glycoproteins: the potential of the sialic acid biosynthetic pathway [J]. J Pharm Sci, 2009, 98(10): 3499–3508.

    Article  CAS  PubMed  Google Scholar 

  2. Varki A. Multiple changes in sialic acid biology during human evolution [J]. Glycoconj J, 2009, 26(3): 231–245.

    Article  CAS  PubMed  Google Scholar 

  3. Adamczyk B, Tharmalingam T, Rudd P M. Glycans as cancer biomarkers [J]. Biochim Biophys Acta, 2012, 1820(9): 1347–1353.

    Article  CAS  PubMed  Google Scholar 

  4. Wang P J. Altered glycosylation in cancer: Sialic acids and sialyltransferases [J]. Cancer Mol, 2005, 1(2): 73–81.

    CAS  Google Scholar 

  5. Schauer R. Achievements and challenges of sialic acid research [J]. Glycoconj J, 2000, 17(7–9): 485–499.

    Article  CAS  PubMed  Google Scholar 

  6. Nie H, Li Y, Sun X L. Recent advances in sialic acid-focused glycomics [J]. J Proteomics, 2012, 75(11): 3098–3112.

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  7. Snovida S I, Rak-Banville J M, Perreault H. On the use of DHB/Aniline and DHB/N,N-dimethylaniline matrices for improved detection of carbohydrates: Automated identification of oligosaccharides and quantitative analysis of sialylated glycans by MALDI-TOF mass spectrometry [J]. J Am Soc Mass Spectrom, 2008, 19(8): 1138–1146.

    Article  CAS  PubMed  Google Scholar 

  8. Harvey D J, Hunter A P, Bateman R H, et al. Relationship between in-source and post-source fragment ions in the matrix-assisted laser desorption (ionization) mass spectra of carbohydrates recorded with reflectron time-of-flight mass spectrometers [J]. Int J Mass Spectrom, 1999, 188(1–2): 131–146.

    Article  CAS  Google Scholar 

  9. Balog C I, Stavenhagen K, Fung W L, et al. N-glycosylation of colorectal cancer tissues: A liquid chromatography and mass spectrometry-based investigation [J]. Mol Cell Proteomics, 2012, 11(9): 571–585.

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  10. Pitt J J, Gorman J J. Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry of sialylated glycopeptides and proteins using 2,6-dihydroxyacetophenone as a matrix [J]. Rapid Commun Mass Spectrom, 1996, 10: 1786–1788.

    Article  CAS  Google Scholar 

  11. Satomaa T, Renkonen O, Helin J, et al. O-glycans on human high endothelial CD34 putatively participating in L-selectin recognition [J]. Blood, 2002, 99(7): 2609–2611.

    Article  CAS  PubMed  Google Scholar 

  12. Fenaille F, Le Mignon M, Groseil C, et al. Combined use of 2,4,6-trihydroxyacetophenone as matrix and enzymatic deglycosylation in organic-aqueous solvent systems for the simultaneous characterization of complex glycoproteins and N-glycans by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry [J]. Rapid Commun Mass Spectrom, 2007, 21(5): 812–815.

    Article  CAS  PubMed  Google Scholar 

  13. Papac D, Wong A, Jones A. Analysis of acidic oligosaccharides and glycopeptides by matrix assisted laser desorption/ionization time-of-flight mass spectrometry [J]. Anal Chem, 1996, 68(18): 3215–3223.

    Article  CAS  PubMed  Google Scholar 

  14. Selman M H, Hoffmann M, Zauner G, et al. MALDI-TOF-MS analysis of sialylated glycans and glycopeptides using 4-chloro-α-cyanocinnamic acid matrix [J]. Proteomics, 2012, 12(9): 1337–1348.

    Article  CAS  PubMed  Google Scholar 

  15. Almeida A, Ferreira J A, Teixeira F, et al. Challenging the limits of detection of sialylated Thomsen-Friedenreich antigens by in-gel deglycosylation and nano-LC-MALDI-TOF-MS [J]. Electrophoresis, 2013, 34(16): 2337–2341.

    Article  CAS  PubMed  Google Scholar 

  16. Mechref Y, Kang P, Novotny M. Solid-phase permethylation for glycomic analysis [J]. Methods Mol Biol, 2009, 534: 53–64.

    CAS  PubMed  Google Scholar 

  17. Desantos-Garcia J L, Khalil S I, Hussein A, et al. Enhanced sensitivity of LC-MS analysis of permethylated N-glycans through online purification [J]. Electrophoresis, 2011, 32(24): 3516–3525.

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  18. Powell A, Harvey D. Stabilization of sialic acids in N-linked oligosaccharides and gangliosides for analysis by positive ion matrix assisted laser desorption ionization mass spectrometry [J]. Rapid Commun Mass Spectrom, 1996, 10(9): 1027–1032.

    Article  CAS  PubMed  Google Scholar 

  19. Chen P, Werner-Zwanziger U, Wiesler D, et al. Mass spectrometric analysis of benzoylated sialooligosaccharides and differentiation of terminal α2→3 and α2→6 sialogalactosylated linkages at subpicomole levels [J]. Anal Chem, 1999, 71(21): 4969–4973.

    Article  CAS  PubMed  Google Scholar 

  20. Shah P, Yang S, Sun S, et al. Mass spectrometric analysis of sialylated glycans with use of solid-phase labeling of sialic acids [J]. Anal Chem, 2013, 85(7): 3606–3613.

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  21. Küster B, Krogh T, Mortz E, et al. Glycosylation analysis of gel-separated proteins [J]. Proteomics, 2001, 1(3): 350–361.

    Article  PubMed  Google Scholar 

  22. Charlwood J, Birrell H, Tolson D, et al. Two-dimensional chromatography in the analysis of complex glycans from transferrin [J]. Anal Chem, 1998, 70(13): 2530–2535.

    Article  CAS  PubMed  Google Scholar 

  23. Harvey D, Bateman R, Bordoli R, et al. Ionisation and fragmentation of complex glycans with a quadrupole time-of-flight mass spectrometer fitted with a matrix-assisted laser desorption/ionisation ion source [J]. Rapid Commun Mass Spectrom, 2000, 14(22): 2135–2142.

    Article  CAS  PubMed  Google Scholar 

  24. Hemmorantaa H, Satomaab T, Blomqvistb M, et al. N-glycan structures and associated gene expression reflect the characteristic N-glycosylation pattern of human hematopoietic stem and progenitor cells [J]. Exp Hematol, 2007, 35(8): 1279–1292.

    Article  Google Scholar 

  25. Satomaa T, Heiskanen A, Leonardsson I, et al. Analysis of the human cancer glycome identifies a novel group of tumor-associated N-acetylglucosamine glycan antigens [J]. Cancer Res, 2009, 69(14): 5811–5819.

    Article  CAS  PubMed  Google Scholar 

  26. Toyoda M, Ito H, Matsuno Y K, et al. Quantitative derivatization of sialic acids for the detection of sialoglycans by MALDI MS [J]. Anal Chem, 2008, 80(13): 5211–5218.

    Article  CAS  PubMed  Google Scholar 

  27. Endo S, Morita M, Ueno M, et al. Fluorescent labeling of a carboxyl group of sialic acid for MALDI-MS analysis of sialyloligosaccharides and ganglioside [J]. Biochem Biophys Res Commun, 2009, 378(4): 890–894.

    Article  CAS  PubMed  Google Scholar 

  28. Handa S, Nakamura K J. Modification of sialic acid carboxyl group of ganglioside [J]. Biochem, 1984, 95(5): 1323–1329.

    CAS  Google Scholar 

  29. Han H H, Ma Y, Wang Z, et al. Derivatization of oligosaccharides and analysis by matrix-assisted laser desorption/ionization-time of flight-mass spectrometry [J]. Chinese J Anal Chem, 2010, 38(1): 307–312.

    Article  CAS  Google Scholar 

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Authors and Affiliations

  1. College of Marine Life Sciences, Ocean University of China, Qingdao, 266003, Shandong, China

    Yanfei Peng

  2. College of Chemistry and Molecular Sciences, Wuhan University, Wuhan, 430072, Hubei, China

    Xiaojuan Xu

Authors
  1. Yanfei Peng
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  2. Xiaojuan Xu
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Correspondence to Yanfei Peng.

Additional information

Foundation item: Supported by the National Natural Science Foundation of China (30800193) and Grant from Centre for International Mobility (CIMO), Finland

Biography: PENG Yanfei, female, Associate professor, Ph. D., research direction: structure and function of carbohydrate and its conjugates.

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Peng, Y., Xu, X. Detection of sialylated N-Linked glycans by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry. Wuhan Univ. J. Nat. Sci. 19, 245–252 (2014). https://doi.org/10.1007/s11859-014-1008-2

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  • DOI: https://doi.org/10.1007/s11859-014-1008-2

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