Abstract
Characterization of structural isomers has become increasingly important and extremely challenging in glycobiology. This communication demonstrates the capability of ion-trap mass spectrometry in conjunction with 157 nm photofragmentation to identify different structural isomers of permethylated N-glycans derived from ovalbumin without chromatographic separation. The results are compared with collision-induced dissociation (CID) experiments. Photodissociation generates extensive cross-ring fragment ions as well as diagnostic glycosidic product ions that are not usually observed in CID MS/MS experiments. The detection of these product ions aids in characterizing indigenous glycan isomers. The ion trap facilitates MSn experiments on the diagnostic glycosidic fragments and cross-ring product ions generated through photofragmentation, thus allowing unambiguous assignment of all of the isomeric structures associated with the model glycoprotein used in this study. Photofragmentation is demonstrated to be a powerful technique for the structural characterization of glycans.
Article PDF
Similar content being viewed by others
References
Varki, A. E. A. Essentials of Glycobiology; Cold Spring Harbor Laboratory Press: New York, 1999.
Campbell, C. T.; Yarema, K. J. Large-scale Approaches for Glycobiology. Genome Biol. 2005, 6, 231–238.
Helenius, A.; Aebi, M. Intracellular Functions of N-Linked Glycans. Science. 2001, 291, 2364–2369.
Rudd, P. M.; Elliott, T.; Cresswell, P.; Wilson, I. A.; Dwek, R. A. Glycosylation and the Immune System. Science 2001, 291, 2370–2376.
Shriver, Z.; Raguram, S.; Sasisekharan, R. Glycomics: A Pathway to a Class of New and Improved Therapeutics. Nat. Rev. Drug Discov. 2004, 3, 863–873.
Bianco, G. A.; Toscano, M. A.; Ilarregui, J. M.; Rabinovich, G. A. Impact of Protein-Glycan Interactions in the Regulation of Autoimmunity and Chronic Inflammation. Autoimmun. Rev. 2006, 5, 349–356.
Varki, A. Biological Roles of Oligosaccharides: All of the Theories Are Correct. Glycobiology 1993, 3, 97–130.
Mechref, Y.; Novotny, M. V. Structural Investigations of Glycoconjugates at High Sensitivity. Chem. Rev. 2002, 102, 321–369.
Reinhold, V. N.; Reinhold, B. B.; Costello, C. E. Carbohydrate Molecular Weight Profiling, Sequence, Linkage and Branching Data: ES-MS and CID. Anal. Chem. 1995, 67, 1772–1784.
Dommon, B.; Muller, D. R.; Richter, W. J. High Performance Tandem Mass Spectrometry for Sequence, Branching and Interglycosidic Linkage Analysis of Peracetylated Oligosaccharides. Biomed. Environ. Mass Spectrom. 1990, 19, 390–402.
Dommon, B.; Muller, D. R.; Richter, W. J. Determination of Interglycosidic Linkages in Disaccharides by High Performance Tandem Mass Spectrometry. Int. J. Mass Spectrom. Ion Processes. 1990, 100, 301–311.
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. 1990, 62, 279–286.
Spengler, B.; Dolce, J. W.; Cotter, R. J. Infrared Laser Desorption Mass Spectrometry of Oligosaccharides: Fragmentation Mechanisms and Isomer Analysis. Anal. Chem. 1990, 62, 1731–1737.
Carroll, J. A.; Ngoka, L.; Mccullough, S.; Gard, E.; Jones, A. D.; Lebrilla, C. B. Quadrupole Fourier Transform Mass Spectrometry of Oligosaccharides. Anal. Chem. 1991, 63, 2526–2529.
Carroll, J. A.; Ngoka, L.; Beggs, C. G.; Lebrilla, C. B. Liquid Secondary Ion Mass Spectrometry/Fourier Transform Mass Spectrometry of Oligosaccharide Anions. Anal. Chem. 1993, 65, 1582–1587.
Harvey, D. J.; Rudd, P. M.; Bateman, R. H.; Bordoli, R. S.; Howes, K.; Hoyes, J. B.; Vickers, R. G. Examination of Complex Oligosaccharides by Matrix-Assisted Laser Desorption/Ionization Mass Spectrometry on Time-of-Flight and Magnetic Sector Instruments. Org. Mass Spectrom. 1994, 29, 753–765.
Harvey, D. J.; Naven, T. J. P.; Kuster, B.; Bateman, R. H.; Green, M. R.; Critchley, G. Comparison of Fragmentation Modes for the Structural Determination of Complex Oligosaccharides Ionized by Matrix-Assisted Laser Desorption/Ionization Mass Spectrometry. Rapid Commun. Mass Spectrom. 1995, 9, 1556–1561.
Visuex, N.; deHoffmann, E.; Dommon, B. Structural Analysis of Permethylated Oligosaccharides by Electrospray Tandem Mass Spectrometry. Anal. Chem. 1997, 69, 3193–3198.
Weiskopf, A. S.; Vouros, P.; Harvey, D. J. Characterization of Oligosaccharide Composition and Structure by Quadrupole Ion Trap Mass Spectrometry. Rapid Commun. Mass Spectrom. 1997, 11, 1493–1504.
Naven, T. J. P.; 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. 1997, 11, 1681–1686.
Harvey, D. J. Matrix-Assisted Laser Desorption/Ionization Mass Spectrometry of Carbohydrates. Mass Spectrom. Rev. 1999, 18, 349–451.
Mechref, Y.; Baker, A. G.; Novotny, M. V. Matrix-Assisted Laser Desorption/Ionization Mass Spectrometry of Neutral and Acidic Oligosaccharides with Collision-Induced Dissociation. Carbohydr. Res. 1998, 313, 145–155.
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. 1998, 70, 4441–4447.
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. 1998, 70, 3053–3059.
Konig, S.; Leary, J. A. Evidence for Linkage Position Determination in Cobalt Co-ordinated Pentasaccharides Using Ion Trap Mass Spectrometry. J. Am. Soc. Mass Spectrom. 1998, 9, 1125–1134.
Solouki, T.; Reinhold, B. B.; Costello, C. E.; O’Malley, M.; Guan, S. H.; Marshall, A. G. Electrospray Ionization and Matrix-Assisted Laser Desorption/Ionization Fourier Transform Ion Cyclotron Resonance Mass Spectrometry of Permethylated Oligosaccharides. Anal. Chem. 1998, 70, 857–864.
Harvey, D. J. Electrospray Mass Spectrometry and Fragmentation of N-Linked Carbohydrates Derivatized at the Reducing Terminus. J. Am. Soc. Mass Spectrom. 2000, 11, 900–915.
Harvey, D. J. Collision-Induced Fragmentation of Underivatized N-Linked Carbohydrates Ionized by Electrospray. J. Mass Spectrom. 2000, 35, 1178–1190.
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. 2000, 14, 2135–2142.
Hunnam, V.; Harvey, D. J.; Priestman, D. A.; Bateman, R. H.; Bordoli, R. S.; Tyldesley, R. Ionization and Fragmentation of Neutral and Acidic Glycosphingolipids with a Q-TOF Mass Spectrometer Fitted with a MALDI Ion Source. J. Am. Soc. Mass Spectrom. 2001, 12, 1220–1225.
Spengler, B.; Kirsch, D.; Kaufmann, R.; Lemoine, J. Structural Analysis of Branched Oligosaccharides Using Post-Source Decay in Matrix-Assisted Laser Desorption Ionization Mass Spectrometry. Org. Mass Spectrom. 1994, 12, 782–787.
Sato, Y.; Suzuki, M.; Nirasawa, T.; Suzuki, A.; Endo, T. Microsequencing of Glycans Using 2-Aminobenzamide and MALDI-TOF Mass Spectrometry: Occurrence of Unique Linkage-Dependent Fragmentation. Anal. Chem. 2000, 72, 1207–1216.
Rouse, J. C.; Strang, A.-M.; Yu, W.; Vath, J. E. Isomeric Differentiation of Asparagine-Linked Oligosaccharides by Matrix-Assisted Laser Desorption-Ionization Postsource Decay Time-of-Flight Mass Spectrometry. Anal. Biochem. 1998, 256, 33–46.
Mechref, Y.; Novotny, M. V.; Krishnan, C. Structural Characterization of Oligosaccharides Using Maldi-TOF/TOF Tandem Mass Spectrometry. Anal. Chem. 2003, 75, 4895–4903.
Mechref, Y.; Kang, P.; Novotny, M. V. Differentiating Structural Isomers of Sialylated Glycans by Matrix-Assisted Laser Desorption/Ionization Time-of-Flight/Time-of-Flight Tandem Mass Spectrometry. Rapid Commun. Mass Spectrom. 2006, 20, 1381–1389.
Spina, E. S. 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. 2004, 18, 392–398.
Morelle, W.; Slomianny, M.-C.; Diemer, H.; Schaeffer, C.; van Dorsselaer, A.; Michalski, J.-C. Fragmentation Characteristics of Neutral N-Linked Glycans Using a MALDI-TOF/TOF Tandem Mass Spectometer. Anal. Chem. 2004, 76, 2343–2354.
Morelle, W.; Slomianny, M.-C.; Diemer, H.; Schaeffer, C.; van Dorsselaer, A.; 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. 2004, 18, 2637–2649.
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. 2007, 79, 3830–3842.
Shi, S. D.-H.; Hendrickson, C. L.; Marshall, A. G.; Seigel, M. M.; Kong, F.; Carter, G. T. Structural Validation of Saccharomicins by High Resolution and High Mass Accuracy Fourier Transform-Ion Cyclotron Resonance-Mass Spectrometry and Infrared Multiphoton Dissociation Tandem Mass Spectrometry. J. Am. Soc. Mass Spectrom. 1999, 10, 1285–1290.
Xie, Y.; Lebrilla, C. B. Infrared Multiphoton Dissociation of Alkali Metal-Coordinated Oligosaccharides. Anal. Chem. 2003, 75, 1590–1598.
Zhang, J.; Schubothe, K.; Li, B.; Russell, S.; Lebrilla, C. B. Infrared Multiphoton Dissociation of O-Linked Mucin-Type Oligosaccharides. Anal. Chem. 2005, 77, 208–214.
Lancaster, K. S.; An, H. J.; Li, B.; Lebrilla, C. B. Interrogation of N-Linked Oligosaccharides Using Infrared Multiphoton Dissociation in FT-ICR Mass Spectrometry. Anal. Chem. 2006, 78, 4990–4997.
Budnik, B. A.; Haselmann, K. F.; Elkin, Y.; Gorbach, V. I.; Zuberev, R. A. Applications of Electron-Ion Dissociation Reactions for Analysis of Polycationic Chitooligosaccharides in Fourier Transform Mass Spectrometry. Anal. Chem. 2003, 75, 5994–6001.
Adamson, J. T.; Hakansson, K. Electron Capture Dissociation of Oligosaccharides Ionized with Alkali, Alkaline Earth, and Transition Metals. Anal. Chem. 2007, 79, 2901–2910.
Thompson, M. S.; Cui, W.; Reilly, J. P. Fragmentation of Singly-charged Peptides by Photodissociation at 157 nm. Angew. Chem. Int. Ed. Engl. 2004, 43, 4791–4794.
Cui, W.; Thompson, M. S.; Reilly, J. P. Pathways of Peptide Ion Fragmentation Induced by Vacuum Ultraviolet Light. J. Am. Soc. Mass Spectrom. 2005, 16, 1384–1398.
Kim, T. Y.; Thompson, M. S.; Reilly, J. P. Peptide Photodissociation at 157 nm in a Linear Ion Trap Mass Spectrometer. Rapid Commun. Mass Spectrom. 2005, 19, 1657–1665.
Devakumar, A.; Thompson, M. S.; Reilly, J. P. Fragmentation of Oligosaccharide Ions with 157 nm Vacuum Ultraviolet Light. Rapid Commun. Mass Spectrom. 2005, 19, 2313–2320.
Devakumar, A.; Mechref, Y.; Kang, P.; Novotny, M. V.; Reilly, J. P. Laser-Induced Photofragmentation of Neutral and Acidic Glycans inside an Ion-Trap Mass Spectrometer. Rapid Commun. Mass Spectrom. 2007, 21, 1452–1460.
Mechref, Y.; Novotny, M. V. Matrix-Assisted Laser Desorption/Ionization Mass Spectrometry of Acidic Glycoconjugates Facilitated by the Use of Spermine as a Co-matrix. J. Am. Soc. Mass Spectrom. 1998, 9, 1293–1302.
Kang, P.; Mechref, Y.; Klouckova, I.; Novotny, M. V. Solid-Phase Permethylation of Glycans for Mass Spectrometric Analysis. Rapid Commun. Mass Spectrom. 2005, 19, 3421–3428.
Devakumar, A.; O’Dell, D. K.; Walker, J. M.; Reilly, J. P. Structural Analysis of Leukotriene C4 isomers Using Collisional Activation and 157 nm Photodissociation. J. Am. Soc. Mass Spectrom. 2007, 19, 14–26.
Payne, A. H.; Glish, G. L. Thermally Assisted Infrared Multiphoton Photodissociation in a Quadrupole Ion Trap. Anal. Chem. 2001, 73, 3542–3548.
Wilson, J. J.; Brodbelt, J. S. MS/MS Simplification by 355 nm Ultraviolet Photodissociation of Chromophore-Derivatized Peptides in a Quadrupole Ion Trap. Anal. Chem. 2007, 79, 7883–7892.
Dommon, B.; Costello, C. E. A Systematic Nomenclature for Carbohydrate Fragmentations in FAB-MS/MS Spectra of Glycoconjugates. Glycoconjugate J. 1988, 5, 397–409.
Spina, E. S. 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. 2004, 18, 392–398.
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. 2004, 76, 2343–2354.
Silva, M. L. C. D.; Stubbs, H. J.; Tamura, T.; Rice, K. G. 1H NMR Characterization of a Hen Ovalbumin Tyrosinamide N-Linked Oligosaccharide Library. Arch. Biochem. Biophys. 1995, 318, 465–475.
Kuster, B.; Naven, T. J. P.; Harvey, D. J. Rapid Approach for Sequencing Neutral Oligosaccharides by Exoglycosidase Digestion and Matrix-Assisted Laser Desorption/Ionization Time-of-Flight Mass Spectrometry. J. Mass Spectrom. 1996, 31, 1131–1140.
North, S.; Birrell, H.; Camilleri, P. Postive and Negative Ion Matrix-Assisted Laser Desorption/Ionization Time-of-Flight Mass Spectrometric Analysis of Complex Glycans Released from Hen Ovalbumin and Derivatized with 2-Aminoacridone. Rapid Commun. Mass Spectrom. 1998, 12, 349–356.
Lattova, E.; Perreault, H.; Krokhin, O. Matrix-Assisted Laser Desorption/Ionization Tandem Mass Spectrometry and Post-Source Decay Fragmentation Study of Phenylhydrazones of N-Linked Oligosaccharides from Ovalbumin. J. Am. Soc. Mass Spectrom. 2004, 15, 725–735.
Lattova, E.; Perreault, H. Profilling of N-Linked Oligosaccharides Using Phenylhydrazine Derivatization and Mass Spectrometry. J. Chromatogr. A. 2003, 1016, 71–87.
Harvey, D. J. Fragmentation of Negative Ions from Carbohydrates: Part 3: Fragmentation of Hybrid and Complex N-Linked Glycans. J. Am. Soc. Mass Spectrom. 2005, 16, 647–659.
Author information
Authors and Affiliations
Corresponding author
Additional information
Published online March 25, 2008
Electronic supplementary material
Rights and permissions
About this article
Cite this article
Devakumar, A., Mechref, Y., Kang, P. et al. Identification of isomeric N-glycan structures by mass spectrometry with 157 nm laser-induced photofragmentation. J Am Soc Mass Spectrom 19, 1027–1040 (2008). https://doi.org/10.1016/j.jasms.2008.03.005
Received:
Revised:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1016/j.jasms.2008.03.005