Abstract
Electron capture dissociation (ECD) has become an alternative method to collision-activated dissociation (CAD) to avoid gas-phase cleavage of post-translational modifications carried by side chains from the peptide backbone. Nonetheless, as illustrated herein by the study of O-glycosylated and O-phosphorylated peptides, the extent of ECD fragmentations may be insufficient to cover the entire peptide sequence and to localize accurately these modifications. The present work demonstrates that the derivatization of peptides at their N-terminus by a phosphonium group improves dramatically and systematically the sequence coverage deduced from the ECD spectrum for both O-glycosylated and O-phosphorylated peptides compared with their native counterparts. The exclusive presence of N-terminal fragments (c-type ions) in the ECD spectra of doubly charged molecular cations simplifies peptide sequence interpretation. Thus, the combination of ECD and fixed charge derivatization appears as an efficient analytical tool for the extensive sequencing of peptides bearing labile groups.
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Huddleston, M. J.; Annan, R. S.; Bean, M. F.; Carr, S. A. Selective Detection of Phosphopeptides in Complex Mixtures by Electrospray Liquid Chromatograph/Mass Spectrometry. J. Am. Soc. Mass Spectrom. 1993, 4, 710–717.
Hanisch, F. G.; Green, B. N.; Bateman, R.; Peter-Katalinic, J. Localization of O-Glycosylation Sites of MUC1 Tandem Repeats by QTOF ESI Mass Spectrometry. J. Mass Spectrom. 1998, 33, 358–362.
Rademaker, G. J.; Pergantis, S. A.; Blok-Tip, L.; Langridge, J. I.; Kleen, A.; Thomas-Oates, J. Mass Spectrometric Determination of the Sites of O-Glycan Attachment with Low Picomolar Sensitivity. Anal. Biochem. 1998, 257, 149–160.
Greis, K. D.; Hayes, B. K.; Comer, F. I.; Kirk, M. K.; Barnes, S.; Lowary, T. L.; Hart, G. W. Selective Detection and Site-Analysis of O-GlcNAc-Modified Glycopeptides by β-Elimination and Tandem Electrospray Mass Spectrometry. Anal. Biochem. 1996, 234, 38–49.
Mirgorodskaya, E.; Hassan, H.; Clausen, H.; Roepstorff, P. Mass Spectrometric Determination of O-Glycosylation Sites Using β-Elimination and Partial Acid Hydrolysis. Anal. Chem. 2001, 73, 1263–1269.
Mirgorodskaya, E.; Roepstorff, P.; Zubarev, R. A. Localization of O-Glycosylation Sites in Peptides by Electron Capture Dissociation in a Fourier Transform Mass Spectrometer. Anal. Chem. 1999, 71, 4431–4436.
Cooper, H.; Håkansson, K.; Marshall, A. G. The Role of Electron Capture Dissociation in Biomolecular Analysis. Mass Spectrometry. Reviews. 2005, 24, 201–222.
Syka, J. E. P.; Coon, J. J.; Schroeder, M. J.; Shabanowitz, J.; Hunt, D. Peptide and Protein Sequence Analysis by Electron Transfer Dissociation Mass Spectrometry. PNAS. 2004, 26, 9528–9533.
Shi, S. D.-H.; Hemling, M. E.; Carr, S. A.; Horn, D. M.; Lindgh, I.; McLafferty, F. W. Phosphopeptide/Phosphoprotein Mapping by Electron Capture Dissociation Mass Spectrometry. Anal. Chem. 2001, 73, 19–22.
Kelleher, N. L.; Zubarev, R. A.; Bush, K.; Furie, B.; Furie, B. C.; McLafferty, F. W.; Walsh, C. T. Localization of Labile Posttranslational Modifications by Electron Capture Dissociation: The Case of γ-Carboxyglutamic Acid. Anal. Chem. 1999, 71, 4250–4253.
Ge, Y.; Lawhorn, B. G.; El Naggar, M.; Strauss, E.; Park, J.-H.; Begley, T. P.; McLafferty, F. W. Top Down Characterization of Larger Proteins (45 kDa) by Electron Capture Dissociation Pass Spectrometry. J. Am. Chem. Soc. 2002, 124, 672–678.
Bogdanov, B.; Smith, R. D. Proteomics by FTICR Mass Spectrometry: Top Down and Bottom Up. Mass Spectrom. Rev. 2005, 24, 168–200.
O’Connor, P. B.; Lin, C.; Cournoyer, J. J.; Pittman, J. L.; Belyayev, M.; Budnik, B. A. Long-Lived Electron Capture Dissociation Product Ions Experience Radical Migration via Hydrogen Abstraction. J. Am. Soc. Mass Spectrom. 2006, 17, 576–585.
Savitski, M. M.; Kjeldsen, F.; Nielsen, M. L.; Zubarev, R. A. Hydrogen Rearrangement to and from Radical z-Fragments in Electron Capture Dissociation of Peptides. J. Am. Soc. Mass Spectrom. 2007, 18, 113–120.
Horn, D. M.; Ge, Y.; McLafferty, F. W. Activated Ion Electron Capture Dissociation for Mass Spectral Sequencing of Larger (42 kDa) Proteins. Anal. Chem. 2000, 72, 4778–4784.
Han, X.; Jin, M.; Breuker, K.; McLafferty, F. W. Extending Top-Down Mass Spectrometry to Proteins with Masses Greater than 200 kDa. Science 2006, 314, 109–112.
Chalmers, M. J.; Håkansson, K.; Johnson, R.; Smith, R.; She, J.; Emmett, M. R.; Marshall, A. G. Protein Kinase A Phosphorylation Characterized by Tandem Fourier Transform Ion Cyclotron Resonance Mass Spectrometry. Proteomics 2004, 4, 970–981.
Håkansson, K.; Chalmers, M. J.; Quinn, J. P.; McFarland, M. A.; Hendrickson, C. L.; Marshall, A. G. Combined Electron Capture and Infrared Multiphoton Dissociation for Multistage MS/MS in a Fourier Transform Ion Cyclotron Resonance Mass Spectrometer. Anal. Chem. 2003, 75, 3256–3262.
Sadagopan, N.; Malone, M.; Watson, J. T. Effect of Charge Derivatization in the Determination of Phosphorylation Sites in Peptides by Electrospray Ionization Collision-activated Dissociation Tandem Mass Spectrometry. J. Mass Spectrom. 1999, 34, 1279–1282.
Czeszak, X.; Morelle, W.; Ricart, G.; Tetaert, D.; Lemoine, J. Localization of the O-Glycosylated Sites in Peptides by Fixed-Charge Derivatization with a Phosphonium Group. Anal. Chem. 2004, 76, 4320–4324.
Caravatti, P.; Allemann, M. The Infinity Cell—A New Trapped-Ion Cell with Radiofrequency Covered Trapping Electrodes for Fourier-Transform Ion-Cyclotron Resonance Mass-Spectrometry. Org. Mass Spectrom. 1991, 26, 514–518.
Gauthier, J. W.; Trautman, T. R.; Jacobson, D. B. Sustained Off-Resonance Irradiation for Collision-Activated Dissociation Involving Fourier Transform Mass Spectrometry: Collision-Activated Dissociation Technique that Emulates Infrared Multiphoton Dissociation. Anal. Chim. Acta. 1991, 246, 211–225.
Tsybin, Y. O.; Ramström, M.; Witt, M.; Baykut, G.; Håkansson, P. Peptide and Protein Characterization by High-Rate Electron Capture Dissociation Fourier Transform Ion Cyclotron Resonance Mass Spectrometry. J. Mass Spectrom. 2004, 39, 719–729.
Zubarev, R. A.; Kelleher, N. L.; McLafferty, F. W. Electron Capture Dissociation of Multiply Charged Protein Cations: A Nonergodic Process. J. Am. Chem. Soc. 1998, 120, 3265–3266.
Cooper, H. J. Investigation of the Presence of b Ions in Electron Capture Dissociation Mass Spectra. J. Am. Soc. Mass Spectrom. 2005, 16, 1932–1940.
Haselmann, K. F.; Schmidt, M. Do b-Ions Occur from Vibrational Excitation Upon H-Desorption in Electron Capture Dissociation?. Rapid Commun. Mass Spectrom. 2007, 21, 1003–1008.
Mormann, M.; Paulsen, H.; Peter-Katalinic, J. Electron Capture Dissociation of O-Glycosylated Peptides: Radical Site-Induced Fragmentation of Glycosidic Bond. Eur. J. Mass Spectrom. 2005, 11, 497–511.
Syrstad, E. A.; Turecek, F. Toward a General Mechanism of Electron Capture Dissociation. J. Am. Soc. Mass Spectrom. 2005, 16, 208–224.
Anusiewicz, I.; Berdys-Kochanska, J.; Skurski, P.; Simons, J. Simulating Electron Transfer Attachment to a Positively Charged Model Peptide. J. Phys. Chem. A. 2006, 110, 1261–1266.
Anusiewicz, I.; Berdys-Kochanska, J.; Simons, J. The Electron Attachment Step in Electron Capture (ECD) and Electron Transfer Dissociation (ETD). J. Phys. Chem. A. 2005, 109, 5801–5813.
Sobczyk, M.; Simons, J. Distance Dependence of Through-Bond Electron Transfer Rates in Electron-Capture and Electron-transfer Dissociation. Int. J. Mass Spectrom. 2006, 253, 274–280.
Sobczyk, M.; Simons, J. The Role of Excited Rydberg States in Electron Transfer Dissociation. J. Phys. Chem. B. 2006, 110, 7519–7527.
Huang, Z. H.; Wu, J.; Gage, D. A.; Watson, J. T. A Picomole-Scale Method for Charge Derivatization of Peptides for Sequence Analysis by Mass Spectrometry. Anal. Chem. 1997, 69, 137–144.
Sadagopan, N.; Watson, J. T. Investigation of the Tris(Trimethoxyphenyl)Phosphonium Acetyl Charged Derivatives of Peptides by Electrospray Ionization Mass Spectrometry and Tandem Mass Spectrometry. J. Am. Soc. Mass Spectrom. 2000, 11, 107–119.
Huang, Z. H.; Shen, T.; Wu, J.; Gage, D. A.; Watson, J. T. Protein Sequencing by Matrix-Assisted Laser Desorption Ionization-Postsource Decay-Mass Spectrometry Analysis of the N-Tris(2,4,6-Trimethoxyphenyl)Phosphine-Acetylated Tryptic Digests. Anal. Biochem. 1999, 268, 305–317.
Roth, K. D. W.; Huang, Z. H.; Sadagopan, N.; Watson, J. T. Charge Derivatization of Peptides for Analysis by Mass Spectrometry. Mass Spectrom. Rev. 1998, 17, 255–274.
Sadagopan, N.; Watson, J. T. Mass Spectrometric Evidence for Mechanisms of Fragmentation of Charge-Derivatized Peptides. J. Am. Soc. Mass Spectrom. 2001, 12, 399–409.
Chen, W.; Lee, P. J.; Shion, H.; Ellor, N.; Gebler, J. C. Improving de Novo Sequencing of Peptides Using a Charged Tag and C-Terminal Digestion. Anal. Chem. 2007, 79, 1583–1590.
Iavarone, A. T.; Paech, K.; Williams, E. R. Effects of Charge and Cationizing Agent on the Electron Capture Dissociation of a Peptide. Anal. Chem. 2004, 76, 2231–2238.
Zubarev, R. A.; Haselmann, K. F.; Budnik, B.; Kjeldsen, F.; Jensen, F. Towards an Understanding of the Mechanism of Electron-Capture Dissociation: A Historical Perspective and modern ideas. Eur. J. Mass Spectrom. 2002, 8, 337–349.
Håkansson, K.; Hudgins, R. R.; Marshall, A. G. Electron Capture Dissociation and Infrared Multiphoton Dissociation of Oligodeoxynucleotide Dications. J. Am. Soc. Mass Spectrom. 2003, 14, 23–41.
Fung, Y. M. E.; Liu, H.; Chan, T.-W. D. Electron Capture Dissociation of Peptides Metalated with Alkaline-Earth Metal Ions. J. Am. Soc. Mass Spectrom. 2006, 17, 757–771.
Liu, H.; Håkansson, K. Electron Capture Dissociation of Tyrosine O-Sulfated Peptides Complexed with Divalent Metal Cations. Anal. Chem. 2006, 78, 7570–7576.
Shaffer, S. A.; Turecek, F. Hydrogentrimethylammonium: A marginally stable hypervalent radical. J. Am. Chem. Soc. 1994, 116, 8647–8653.
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Published online April 25, 2007
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Chamot-Rooke, J., van der Rest, G., Dalleu, A. et al. The combination of electron capture dissociation and fixed charge derivatization increases sequence coverage for O-glycosylated and O-phosphorylated peptides. J Am Soc Mass Spectrom 18, 1405–1413 (2007). https://doi.org/10.1016/j.jasms.2007.04.008
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DOI: https://doi.org/10.1016/j.jasms.2007.04.008