Comparative ESI FT-MS and MALDI-TOF structural analyses of representative human N-linked glycans
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Zuzana Pakanová
,
Marek Nemčovič
Abstract
Modern glycan analysis is primarily based on mass spectrometry, where instruments based on electrospray or matrix-assisted laser desorption ionization are currently the most frequently used. In the present study, electrospray ionization (ESI) coupled with a high-resolution Fourier transform mass spectrometer (LTQ Orbitrap) and matrix-assisted laser desorption/ionization (MALDI) coupled with a time-of-flight (TOF/TOF) detector were used to analyze two N-glycan standards with intact free reducing ends (disialo biantennary and asialo triantennary) and representative PA-labeled human serum N-glycan structures isolated by hydrophilic interaction anion-exchange chromatography (HIAX), confirmed by 1H NMR analysis and consequently compared with the ProteinScape Glycome database. Different combinations of ion sources with fragmentation devices results in various fragmentation patterns and adducts. Also, the effect of sample derivatization on the acquired signals is discussed. Compared to the MALDI technique, free glycans did not lose labile sialic acids easily in the ESI source. On the other hand, fluorescent PA-labeling leads to improved core fragmentation and signal intensities; linkage-specific ethyl esterification leads to reduced adduct and fragment formation and enhanced stability of sialic acids in the MALDI ion source. Thereby, both methods have their advantages and disadvantages in terms of detection, fragmentation and robustness.
References
Arnal-Estapé, A., & Nguyen, D. X. (2015). Sweets for a bitter end: Lung cancer cell-surface protein glycosylation mediates metastatic colonization. Cancer Discovery, 5, 109-111. DOI: 10.1158/2159-8290.cd-15-0013.10.1158/2159-8290.CD-15-0013Search in Google Scholar PubMed PubMed Central
Debeire, P., Montreuil, J., Moczar, E., Van Halbeek, H., & Vliegenthart, J. F. G. (1985). Primary structure of two major glycans of bovine fibrinogen. European Journal of Biochemistry, 151, 607-611. DOI: 10.1111/j.1432-1033.1985.tb09147.x.10.1111/j.1432-1033.1985.tb09147.xSearch in Google Scholar PubMed
Debray, H., Fournet, B., Montreuil, J., Dorland, L., & Vliegenthart, J. F. G. (1981). Structure determination by 360- MHz 1H-NMR spectroscopy and methylation analysis of a biantennary glycan of the N-acetyllactosaminic type isolated from rat-liver plasma membrane. European Journal of Biochemistry, 115, 559-563. DOI: 10.1111/j.1432-1033.1981.tb06239.x.10.1111/j.1432-1033.1981.tb06239.xSearch in Google Scholar PubMed
Durand, G., & Seta, N. (2000). Protein glycosylation and diseases: Blood and urinary oligosaccharides as markers for diagnosis and therapeutic monitoring. Clinical Chemistry, 46, 795-805.10.1093/clinchem/46.6.795Search in Google Scholar
Freeze, H. H., & Aebi, M. (2005). Altered glycan structures: the molecular basis of congenital disorders of glycosylation. Current Opinion in Structural Biology, 15, 490-498. DOI: 10.1016/j.sbi.2005.08.010.10.1016/j.sbi.2005.08.010Search in Google Scholar PubMed
Giménez, E., Balmaña, M., Figueras, J., Fort, E., de Bolós, C., Sanz-Nebot, V., Peracaula, R., & Rizzi, A. (2015). Quantitative analysis of N-glycans from human alfa-acidglycoprotein using stable isotope labeling and zwitterionic hydrophilic interaction capillary liquid chromatography electrospray mass spectrometry as tool for pancreatic disease diagnosis. Analytica Chimica Acta, 866, 59-68. DOI: 10.1016/j.aca.2015.02.008.10.1016/j.aca.2015.02.008Search in Google Scholar PubMed
Hollingsworth, M. A., & Swanson, B. J. (2004). Mucins in cancer: protection and control of the cell surface. Nature Reviews Cancer, 4, 45-60. DOI: 10.1038/nrc1251.10.1038/nrc1251Search in Google Scholar PubMed
Novak, J., Tomana,M., Shah, G. R., Brown, R.,&Mestecky, J. (2005). Heterogeneity of IgG glycosylation in adult periodontal disease. Journal of Dental Research, 84, 897-901. DOI: 10.1177/154405910508401005.10.1177/154405910508401005Search in Google Scholar PubMed
Novotny, M. V., & Mechref, Y. (2005). New hyphenated methodologies in high-sensitivity glycoprotein analysis. Journal of Separation Science, 28, 1956-1968. DOI: 10.1002/jssc.200500258.10.1002/jssc.200500258Search in Google Scholar PubMed PubMed Central
Paschinger, K., Hykollari, A., Razzazi-Fazeli, E., Greenwell, P., Leitsch, D., Walochnik, J., & Wilson, I. B. H. (2012). The N-glycans of Trichomonas vaginalis contain variable core and antennal modifications. Glycobiology, 22, 300-313. DOI: 10.1093/glycob/cwr149.10.1093/glycob/cwr149Search in Google Scholar PubMed PubMed Central
Reiding, K. R., Blank, D., Kuijper, D. M., Deelder, A. M., & Wuhrer, M. (2014). High-throughput profiling of protein N-glycosylation by MALDI-TOF-MS employing linkagespecific sialic acid esterification. Analytical Chemistry, 86, 5784-5793. DOI: 10.1021/ac500335t.10.1021/ac500335tSearch in Google Scholar PubMed
Schedin-Weiss, S., Winblad, B., & Tjernberg, L. O. (2014). The role of protein glycosylation in Alzheimer disease. The FEBS Journal, 281, 46-62. DOI: 10.1111/febs.12590.10.1111/febs.12590Search in Google Scholar PubMed
Institute of Chemistry, Slovak Academy of Sciences
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