Abstract
A new multifunctional oligosaccharide label with a 1o amino-group was synthesized and characterized. The oligosaccharide label was introduced into several neutral oligosaccharides by reductive amination, and the derivatives were analyzed by matrix-assisted laser desorption/ionization (MALDI) time-of-flight (TOF) and by electrospray ionization (ESI) mass spectrometry. It was demonstrated that the labeling reaction was satisfactory, and that as little as 50 pmol of starting material could be efficiently labeled with minimal loss to side reactions. A mixture of high-mannose N-glycans released from ribonuclease B was labeled. The label did not appear to interfere with structural characterization of the oligosaccharides by mass spectrometry. N-quaternization of the labeled oligosaccharides resulted in significantly increased sensitivity of detection with as little as 100 fmol on the probe detected. Deuterium coding of labeled oligosaccharide mixtures and relative abundance of mixture components was investigated. A protocol for the chromatographic separation of mixtures of labeled oligosaccharides by HPLC was developed and is reported here.
Article PDF
Similar content being viewed by others
Avoid common mistakes on your manuscript.
References
Hase, S. Precolumn derivatization for chromatographic and electrophoretic analyses of carbohydrates. J. Chromatogr. A 1996, 720, 173–182.
Harvey, D. J. Matrix-assisted laser desorption/ionization mass spectrometry of carbohydrates and glycoconjugates. Int. J. Mass Spectrom. 2003, 226, 1–35.
Harvey, D. J. Matrix-assisted laser desorption/ionization mass spectrometry of carbohydrates. Mass Spectrom. Rev. 1999, 18, 349–450.
Militsopoulou, M.; Lamari-Fotini, N.; Hjerpe, A.; Karamanos Nikos, K. Determination of twelve heparin- and heparan sulfate-derived disaccharides as 2-aminoacridone derivatives by capillary zone electrophoresis using ultraviolet and laser-induced fluorescence detection. Electrophoresis 2002, 23, 1104–1109.
Honda, S.; Okeda, J.; Iwanaga, H.; Kawakami, S.; Taga, A.; Suzuki, S.; Imai, K. Ultramicroanalysis of reducing carbohydrates by capillary electrophoresis with laser-induced fluorescence detection as 7-nitro-2,1,3-benzoxadiazole-tagged N-methylglycamine derivatives. Anal. Biochem. 2000, 286, 99–111.
Okafo, G.; Langridge, J.; North, S.; Organ, A.; West, A.; Morris, M.; Camilleri, P. High-performance liquid chromatographic analysis of complex N-linked glycans derivatized with 2-aminoacridone. Anal. Chem. 1997, 69, 4985–4993.
Charlwood, J.; Birrell, H.; Gribble, A.; Burdes, V.; Tolson, D.; Camilleri, P. A probe for the versatile analysis and characterization of N-linked oligosaccharides. Anal. Chem. 2000, 72, 1453–1461.
Ahn, Y. H.; Yoo, J. S. Efficient analysis of oligosaccharide-malononitrile derivatives by on-line capillary liquid chromatography/electrospray ionization mass spectrometry. Rapid Commun. Mass Spectrom. 1999, 13, 855–859.
Lemoine, J.; Cabanes-Macheteau, M.; Bardor, M.; Michalski, J.-C.; Faye, L.; Lerouge, P. Analysis of 8-aminonaphthalene-1,3,6-trisulfonic acid labeled N-glycans by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry. Rapid Commun. Mass Spectrom. 2000, 14, 100–104.
Suzuki, H.; Mueller, O.; Guttman, A.; Karger, B. L. Analysis of 1-aminopyrene-3,6,8-trisulfonate-derivatized oligosaccharides by capillary electrophoresis with matrix-assisted laser desorption/ionization time-of-flight mass spectrometry. Anal. Chem. 1997, 69, 4554–4559.
Turecek, F. Mass spectrometry in coupling with affinity capture-release and isotope-coded affinity tags for quantitative protein analysis. J. Mass Spectrom. 2002, 37, 1–14.
Patton, W. F.; Schulenberg, B.; Steinberg, T. H. Two-dimensional gel electrophoresis; better than a poke in the ICAT? Curr. Opin. Biotechnol. 2002, 13, 321–328.
Patton, W. F. Detection technologies in proteome analysis. Analytical technologies in the biomedical and life sciences. J. Chromatogr. B 2002, 771, 3–31.
Mo, W.; Sakamoto, H.; Nishikawa, A.; Kagi, N.; Langridge, J. I.; Shimonishi, Y.; Takao, T. Structural characterization of chemically derivatized oligosaccharides by nanoflow electrospray ionization mass spectrometry. Anal. Chem. 1999, 71, 4100–4106.
Harvey, D. J. Collision-induced fragmentation of underivatized N-linked carbohydrates ionized by electrospray. J. Mass Spectrom. 2000, 35, 1178–1190.
Zeleny, R.; Altmann, F.; Praznik, W. Structural characterization of the N-linked oligosaccharides from tomato fruit. Phytochemistry 1999, 51, 199–210.
Mechref, Y.; Novotny, M. V. Structural investigations of glycoconjugates at high sensitivity. Chem. Rev. 2002, 102, 321–369.
Kennedy, R. T.; German, I.; Thompson, J. E.; Witowski, S. R. Fast analytical-scale separations by capillary electrophoresis and liquid chromatography. Chem. Rev. 1999, 99, 3081–3131.
Rothenberg, B. E.; Hayes, B. K.; Toomre, D.; Manzi, A. E.; Varki, A. Biotinylated diaminopyridine: An approach to tagging oligosaccharides and exploring their biolog. Proceedings of the National Academy of Sciences of the United States of America; 1993; pp 11939–11943.
Toomre, D. K.; Varki, A. Advances in the use of biotinylated diaminopyridine (BAP) as a versatile fluorescent tag for oligosaccharides. Glycobiology 1994, 4, 653–663.
Varki, A.; Rothenberg, B. E. Novel Fluorescent Tagging Agents, Patent Corporation Treaty (PCT) International Property Organization, World Intellectual Property Organization, 1994, pp 38.
Shinohara, Y.; Sota, H.; Gotoh, M.; Hasebe, M.; Tosu, M.; Nakao, J.; Hasegawa, Y.; Shiga, M. Bifunctional labeling reagent for oligosaccharides to incorporate both chromophore and biotin groups. Anal. Chem. 1996, 68, 2573–2579.
Love, K. R.; Seeberger, P. H. Carbohydrate arrays as tools for glycomics. Angew. Chem. Int. Ed. 2002, 41, 3583–3586.
Leteux, C.; Childs, R. A.; Chai, W.; Stoll, M. S.; Kogelberg, H.; Feizi, T. Biotinyl-L-3-(2-naphthyl)-alanine hydrazide derivatives of N-glycans: Versatile solid-phase probes for carbohydrate-recognition studies. Glycobiology 1998, 8, 227–236.
Broberg, S.; Broberg, A.; Duus, J. O. Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry of oligosaccharides derivatized by reductive amination and N,N-dimethylation. Rapid Commun. Mass Spectrom. 2000, 14, 1801–1805.
An, H. J.; Franz, A. H.; Lebrilla, C. B. Improved capillary electrophoretic separation and mass spectrometric detection of oligosaccharides. J. Chromatogr. A 2003, 1004, 121–129.
Domon, B.; Costello, C. E. A systematic nomenclature for carbohydrate fragmentations in FAB-MS/MS spectra of glycoconjugates. Glycoconj. J. 1988, 5, 397–409.
Garlick, R. K.; Giese, R. W. Avidin binding of radiolabeled biotin derivatives. J. Biol. Chem. 1988, 263, 210–215.
Plummer, T. H., Jr.; Elder, J. H.; Alexander, S.; Phelan, A. W.; Tarentino, A. L. Demonstration of peptide: N-glycosidase F activity in endo-b-N-acetylglucosaminidase F preparations. J. Biol. Chem. 1984, 259, 10700–10704.
An, H. J.; Peavy, T. R.; Hedrick, J. L.; Lebrilla, C. B. Determination of N-glycosylation sites and site heterogeneity in glycoproteins. Anal. Chem. 2003, 75, 5628–5637.
Plummer, T. H., Jr.; Tarentino, A.; Maley, F. The glycopeptide linkage of ribonuclease B. J. Biol. Chem. 1968, 243, 5158–5164.
Bahl, O. P.; Agrawal, K. M. L. Glycosidases of Phaseolus vulgaris. I. Isolation and characterization of b-N-acetylglucosaminidase. J. Biol. Chem. 1968, 243, 98–102.
Tarentino, A. L.; Plummer, T. H., Jr.; Maley, F. Release of intact oligosaccharides from specific glycoproteins by endo-b-N-acetylglucosaminidase H. J. Biol. Chem. 1974, 249, 818–824.
Takahashi, N. Demonstration of a new amidase acting on glycopeptides. Biochem. Biophys. Res. Commun. 1977, 76, 1194–1201.
Takahashi, N.; Nishibe, H. Some characteristics of a new glycopeptidase acting on aspartylglycosylamine linkages. J. Biochem. Tokyo, Japan 1978, 84, 1467–1473.
Author information
Authors and Affiliations
Corresponding author
Additional information
Published online January 9, 2006
Rights and permissions
About this article
Cite this article
Hsu, J., Chang, S.J. & Franz, A.H. MALDI-TOF and ESI-MS analysis of oligosaccharides labeled with a new multifunctional oligosaccharide tag. The official journal of The American Society for Mass Spectrometry 17, 194–204 (2006). https://doi.org/10.1016/j.jasms.2005.10.010
Received:
Revised:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1016/j.jasms.2005.10.010